Table of Content

2020, Volume 51 Issue 1
06 February 2020 Previous Issue    Next Issue
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Technology and Methodology

Histology,Embryology and Developmental Biology
Anthropology
Anatomy
Neurobiology
Cancer Biology
Bioengineering


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Observation of tissue slice staining with purple BL and blue B2R SHAO Jin-hui FENG Gui-hua 2020, 51 (1):  132-134.  doi: 10.16098/j.issn.0529-1356.2020.01.023 Abstract ( )   PDF (1744KB) ( )    Objective To find out much more simply prepared, effective and abundant tissue slice dye.  Methods Totally 27 kinds of directly used dress dye were tested to make mouse liver slice stained. Among them purple BL and blue B2R made hepatic structure including central vein and hepatic nucleus shown clearly. Then, the heart, kidney and testis slices of mouse were stained by purple BL and blue B2R.  Results The solution preparation of Purple BL and blue B2R was just dissolved in distilled water in five minutes, so they were very much simple and timesaving.After the heart, kidney and testis slices of mouse were stained by purple BL and blue B2R,the stained tissue structure including cardiac muscle cross striation,kidney capsule cavity and spermatogenic cells is clear and similar to tissue stained by HE.  Conclusion The effective dress dye purple BL and blue B2R are much more simply prepared and abundant than hematoxylin. So, it can be partly replaced by purple BL and blue B2R. References | Related Articles | Metrics

Technology and Methodology

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Improvement on the methods about the primary culture of rat brain microvascular endothelial cells and identification TANG Yuan-yu MA Hua-gen LIU Zhao-de 2020, 51 (1):  135-138.  doi: 10.16098/j.issn.0529-1356.2020.01.024 Abstract ( )   PDF (3471KB) ( )   Objective To improve the method  about the primary culture of rat brain microvascular endothelial cells in vitro.  Methods The SD rats aged from 4 to 6 weeks were chosen as research object. After craniotomy, washing and cutting, sieving, density gradient concentration of BSA, digestion of type Ⅱ collagenase and collagenase dispersive enzyme twice, the primary culture was carried out. The target cells were indentified by morphological abservation and immunocytochemical staining of  facter Ⅷ.  Results Cultured for 12 to 24 hours，the cells in vitro migrated outward from the microvascular section. The cells appeared polygonal-shaped，and proliferated in a clustered monolayer. the cell growth density reached 70％-80％ of the bottle bottom after 3 days，and arranged like cobbles. The correlation antigen of Ⅷ factor was positive，they reached confluence with over purity 99％.  Conclusion The method  is available that can successfully separate and cultivate microvascular endothelial cells of rat brains in vitro. References | Related Articles | Metrics

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Application of new methods on the study of ancient human remains in China in recent years YANG Shang-bing CHEN Hui-min DUAN Wan-qi LI Hai-jun CHEN Feng DAI Cheng-ping 2020, 51 (1):  139-149.  doi: 10.16098/j.issn.0529-1356.2020.01.025 Abstract ( )   PDF (716KB) ( )   In the past decade, in addition to using traditional naked-eye morphological analysis, two-dimensional measurement and data comparison, more and more scholars placed great emphasis on the application of new technologies available and supported the study of ancient human remains from a multi-disciplinary perspective. Through outlining recent advances in the application of new techniques on research of ancient human remains, this paper briefly summarized the characteristics of present research and gave an outlook on future research of ancient human remains in China. References | Related Articles | Metrics

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Advances in research on the suboccipital muscles PEI Fei CHEN Guo-ping WANG Yan 2020, 51 (1):  150-152.  doi: 10.16098/j.issn.0529-1356.2020.01.026 Abstract ( )   The suboccipital muscle group has been paid more and more attention by the rehabilitation community. Its special junction and special structure make many diseases and symptoms have important association with it. This article will introduce the research progress of suboccipital muscle group from four parts: anatomy, physiological function, common cause of injury and clinical application. References | Related Articles | Metrics

Histology,Embryology and Developmental Biology

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Polypeptide compound urantide inhibiting expression of typeⅠcollagen in rat heart of atherosclerosis YANG Shu-han WANG Bo SUN Mei-sha CAO Yu-guang WANG Yi-ning CUI Hai-peng ZHAO Juan 2020, 51 (1):  103-108.  doi: 10.16098/j.issn.0529-1356.2020.01.018 Abstract ( )   PDF (4224KB) ( )   Objective To observe the effect of the peptide compound urantide on the expression of type Ⅰ collagen (Col Ⅰ) in the heart tissue of rats with atherosclerosis (As), and to explore its mechanism of prevention and treatment of heart damage in As rats. Methods Sixty healthy male 3-week-old SPF Wistar rats were selected. The As model was established by intraperitoneal injection of vitamin D3 (VD3) to damage the arterial intima and high-fat diet. They were randomly divided into normal group, As model group, simvastatin group and urantide (3 days, 7 days, 14 days) groups. HE staining and Masson trichrome staining were used to observe the morphology and collagen fiber expression of rat hearts. Immunohistochemistry, Western blotting and Real-time PCR were used to detect the expression of Col Ⅰ protein and gene in rat heart.  Results Compared with the normal group, pathological phenomena such As myocardial cell degeneration, intercellular infiltration of a large number of neutrophils, scattered foam cells and hyperemia and hemorrhage were observed in the heart tissues of the As model group. Meanwhile, collagen fibers increased, and the gene and protein expression levels of Col Ⅰ increased. Compared with the As model group, the cardiac pathological phenomena were effectively alleviated after the treatment with urantide. With the extension of the administration time, the collagen fibers decreased, and the gene and protein expression levels of Col Ⅰ were gradually dow-regulated, especially the effect was the best when the drug was given for 14 days.  Conclusion Urantide can inhibit the expression of Col Ⅰ in As heart to reduce myocardial interstitial damage, and has a protective effect on the heart of As rats. References | Related Articles | Metrics

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Effects of hydrogen sulfide on pulmonary vascular remodeling and its inhibitors in rats with pulmonary hypertension HUANG Chu-sheng QIN Jia-jin TAN Xia-xian HUANG Hong-jian XIAN Lei 2020, 51 (1):  109-113.  doi: 10.16098/j.issn.0529-1356.2020.01.019 Abstract ( )   PDF (1174KB) ( )   Objective To explore the effects of hydrogen sulfide on pulmonary vascular remodeling and its inhibitors in rats with pulmonary hypertension（PH）.  Methods Thirty male SD rats were randomly divided into control group (10 rats), model group (10 rats) and H2S intervention group (10 rats), PH model was induced by Lilium Wilfordii in model group, on the basis of model group, rats in H2S intervention group were injected with NaHS (56 μmol/kg) intraperitoneally, while rats in control group were injected with normal saline at the same dose. Four weeks later, the hemodynamic parameters were measured, the right ventricular hypertrophy index (RVHI) was calculated, the pathological changes of pulmonary vessels were detected by HE staining, and the expressions of p38 and c-Jun N-terminal kinase(JNK) proteins in the mitogen-activated protein kinase (MAPK) family were detected by Western blotting and Real-time PCR.  Results There were significant differences in hemodynamics, RVHI, wall thickness as a percentage of vessel diameter(WT)%, pulmonary vessel wall area as a percentage of vascular cross-sectional area(WA)%, p38 and JNK in each group (P<0.05). The expression levels of MSAP, MPAP, RVHI, WT%, WA%, p38 mRNA and JNK mRNA in the model group and H2S intervention group were significantly higher than those in the control group (P<0.05), while the levels of MSAP, MPAP, RVHI, WT%, WA%, p38 mRNA and JNK mRNA in H2S intervention group were significantly lower than those in the model group (P<0.05). The pulmonary artery morphology showed that the wall thickness and lumen stenosis of the model group and the H2S intervention group increased compared with the control group, but the lumen thickness and lumen stenosis of the H2S intervention group were significantly reduced compared with the model group; Western blotting showed that the expressions of p38 and JNK in model group and H2S intervention group were higher than those in control group, while the expressions of p38 and JNK in H2S intervention group were lower than those in model group.  Conclusion H2S can improve hemorheology, right ventricular hypertrophy index, alleviate pulmonary artery wall thickening and lumen stenosis, and inhibit pulmonary vascular remodeling in PH rats. Its mechanism may be related to the down-regulation of JNK and p38 protein expression in MAPK signaling pathway by H2S. References | Related Articles | Metrics

Anthropology

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Limb characteristics of Yangshao residents in Wanggou site in Zhengzhou ZHOU Ya-wei ZHOU Bei GU Wan-fa 2020, 51 (1):  114-123.  doi: 10.16098/j.issn.0529-1356.2020.01.020 Abstract ( )   PDF (1342KB) ( )   Objective To research the physical development of ancient inhabitants in the Neolithic Age.  Methods Referring to Anthropometric Method ,the main projects including humerus, tibia and femur of 108 human bone specimens unearthed from Wanggou site have been measured respectively. The limb bone index, sexual dimorphism and height have been compared and analyzed by Excel software.  Results The overall development level of humerus and femur was higher than that of the control group in the same period and weaker than that of the Inner Mongolian group. The development of the middle femur was weaker than that of the control groups, with less muscle attachment. The difference in the brawniness of humerus between the genders was statistically significant (t=0.811, P>0.05), while the difference in the brawniness of femur was just the opposite (t=0.63, P>0.05). The maximum variation range of the male femur was 40.6 to 49.4 cm, which was at a moderate low level in the control group. The average height of male was 166.22 cm, ranging from 164.47-167.68 cm. The average height of female was 158.06 cm, ranging from 154.02-160.34 cm；There was a significant difference in height between the genders (t=4.96, P<0.01).The height of male is close to the south China group, lower than the comparison group in the same region and the north region, and that of female is at a higher level in the south region. The sexual dimorphism index of Wanggou formation was 5.7 %.  Conclusion The humerus, tibia and femur of the male in Wanggou group are stronger than that of the female, while the upper limbs of the male were stronger than that of the female, and there was no significant difference in the strength of the lower limbs. We suggest that there is no significant difference in nutrition between male and female of Wanggou site during their growth. References | Related Articles | Metrics

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Handgrip strength of Tibetan adult in Lhasa YAO Jie WEN You-feng CHEN Ying SONG Jia LIU Ying-ying 2020, 51 (1):  124-127.  doi: 10.16098/j.issn.0529-1356.2020.01.021 Abstract ( )   PDF (664KB) ( )   Objective To measure the handgrip strength of Tibetan adults in Lhasa, and to explore its changing trends and characteristics. This provides a basis for the determination of the Tibetan grip strength reference value.  Methods China (Xiangshan) CAMRY EH101 electronic gripper measured the grip strength of healthy Tibetan adults aged 20-70 in Chengguan District of Lhasa City. There were a total of 760 respondents (310 males and 450 females). After stratifying them by age and gender, they analyzed their changes.  Results The average handgrip strength of male Tibetan adults aged 20-69 in Lhasa was (34.50±8.36) kg. The median male grip strength was 35 kg. The grip strength reached the peak (39.87 kg) in the 20- group. From the 40- group, the grip strength value decreased with age. The average grip strength of female Tibetan adults was (21.54±5.73) kg. The median female grip strength is 22 kg. There was no significant difference in grip strength between the ages of 20- and 40-.The grip strength reached a peak (23.41 kg) in the 40- group. From the 50- group, the grip strength value decreased with age. The average grip strength of Tibetan adults in Lhasa was (26.83±9.41) kg, and the median grip strength was 26 kg. The average grip strength of males in the same age group was higher than that of females. The hand grip strength decreased with increasing altitude (P<0.05).  Conclusion There is a difference in gender and age in hand grip, and it is also affected by altitude. References | Related Articles | Metrics

Anatomy

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Modified braclial plexus block anesthesia LIU Zhang-yu LI Ying-yun 2020, 51 (1):  62-65.  doi: 10.16098/j.issn.0529-1356.2020.01.011 Abstract ( )   PDF (1033KB) ( )   Objective To analyze the influence of topographic anatomy on brachial plexus block anesthesia, and to provide reference for clinical research and application.  Methods Total of 20 cadavers were selected and located through the modified method  of inferior brachial plexus block of the coracoid process. The nerve blocking points were marked with blue dye, and the nerve concentration sites and marking points were dissected and exposed at the same time. The anatomical relationship between nerve concentration points and body surface markers were explored.  Results The distances from the upper margin of the left centralization to the midpoint of the subclavian border, the inferior margin of the sternoclavicular joint, the shoulder peak and the inferior edge of the coracoid process were (3.62±0.24) cm, (10.39±0.25) cm, (6.67±0.18) cm and (2.80±0.19) cm, respectively. The distances from the upper edge of the right centralization to the midpoint of the subclavian border, the inferior margin of the sternoclavicular joint, the shoulder peak and the inferior edge of the coracoid process were (4.24±0.27) cm, (11.10±0.28) cm, (6.35±0.19) cm and (3.03±0.15) cm, respectively.  Conclusion The application of anatomy improves the accuracy of brachial plexus block, thus improves the effect of brachial plexus block anesthesia, and provides a powerful reference for clinical research and application. References | Related Articles | Metrics

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Effect of double chain method on the closure of orthodontic teeth ZHAO Chan-yuan WANG Yang-yang WANG Mei-yan WANG Jing HUO Feng WANG Peng 2020, 51 (1):  66-71.  doi: 10.16098/j.issn.0529-1356.2020.01.012 Abstract ( )   PDF (1690KB) ( )   Objective To study the safety and effectiveness of the double chain method  on the closure of the space of the teeth.  Methods Totally 72 cases of different maxillary and mandible dentition space were included in the study. The pulp vitality was measured before and after treatment, the maxillary and mandible first molar teeth. All values were compared to assess the safety of dental pulp vitality. There were 36 cases in tooth extraction group and non-tooth extraction group.The observation period was 4 weeks,the changes of the alveolar space between maxillary and mandible teeth in adult and minor groups were measured before and after treatment. The difference of the total groove spacing before and after treatment were calculated and compared, to assess the effectiveness of space closure.  Results In all 72 patients, there was no significant difference in the vitality of the pulp before and after treatment (P>0.05). Compared to before treatment,the sum of the distance between the upper and lower mandibular groove using the double chain method was significantly reduced for both adult and juvenile patients in the four week tooth extraction group and the nontooth extraction group,the difference was statistically significant (P<0.05).  Conclusion Double chain closed teeth space does not cause pulp degeneration and necrosis. It is safe in clinical practice. Double chain method  is effective, and can simplify the clinical operation. References | Related Articles | Metrics

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Application of 3D digital reconstruction technique in adult sinus CT based volume AN Feng-yang SUN Zhong-wu WANG Yu-hui BAI Ming-yu SUN Jian-bin ZHU Li 2020, 51 (1):  72-77.  doi: 10.16098/j.issn.0529-1356.2020.01.013 Abstract ( )   PDF (2161KB) ( )   Objective To calculate the volume of maxillary sinus and sphenoid sinus of adults in China by CT, and to analyze the differences of gender in the adult maxillary sinus and sphenoid sinus.  Methods Ninety-three CT images of sinus were randomly selected from Peking University Third Hospital. The digital model of maxillary sinus and sphenoid sinus was reconstructed by Mimics17.0 3D reconstruction software. And the sinus volume was obtained by the three-dimensional digital model.  Results The average volume of the right maxillary sinus in the male was (16.69±6.90) ml, and the average volume of the maxillary sinus in the left side of the male was (16.88±7.12) ml. The average volume of the right sphenoid sinus of male was (4.62±2.99) ml, and the average volume of the left sphenoid sinus of male was (5.58±3.72) ml. The average volume of the right maxillary sinus in female was (11.39±5.63) ml, and the average volume of the left maxillary sinus in female was (11.37±5.38) ml. The average volume of the right sphenoid sinus of female was (4.12±2.53) ml, and the average volume of the left sphenoid sinus of female was (3.32±2.30) ml. The left sphenoid sinus and bilateral maxillary sinus volume had gender differences.  Conclusion Three-dimensional reconstruction can quickly obtain a three-dimensional model of the sinus, accurately calculate the sinus volume, and indirectly reflect gender through the sinus volume. It provides a certain reference for further understanding of sinus anatomy. References | Related Articles | Metrics

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Image anatomy of percutaneous retrograde screw fixation of acetabular posterior column YU Ke-quan GAO Shi-chang CHEN Jian-fei WU Min 2020, 51 (1):  78-85.  doi: 10.16098/j.issn.0529-1356.2020.01.014 Abstract ( )   PDF (3414KB) ( )   Objective To investigate the position, direction and fixation range of retrograde screw of the posterior column of acetabulum based on CT data.  Methods Totally 100 cases (50 males, 50 females) of normal adult pelvic CT data were collected from the First Affilated Hospital of Chongqing Medical University. The pelvis was reconstructed by Mimics 17.0 software and imported into Geomagic Studio 2015 software. The virtual screw was placed in the posterior column from the ischial tuberosity to the iliac fossa, the maximum diameter, the entrance point, the exit point，the direction, safe fixation inclination angles of the screw were measured. The range of the screw fixing，the best fluoroscopic views and the easy-to-penetrate site of the screw in the posterior column were assessed.  Results The safe corridor the posterior column of the acetabulum was similar to the “triangular prism” shape. The entrance point was located on the midline of the medial edge and lateral edge of the ischial tuberosity, which were (12.99±1.99) mm in male and (13.26±2.58) mm in female from farthest end of the ischial tuberosity. There were no stitistical significant differences between male and female(P>0.05). The exit points was located at the iliac fossa, which were (23.65±2.42)mm in male and (24.94±2.39) mm in female to the ipsilateral anterior sacroiliac joint line,and (19.33±2.60)mm in male and (17.63±2.00) mm in female to the arcus marginalis. The maximum diameters of screw were (17.21±1.41) mm in male and (15.54±1.51) mm in female. The angles between screw and coronal plane were (15.00±4.92)° in male and (12.94±4.72)° in female; the angles between screw and sagittal plane were (10.52±3.04)° in male and (7.72±2.99)° in female. The abore data had statistially significant differences between male and female (P<0.05). Percutaneous retrograde screw could fix the acetabular posterior column fracture that below the 4.0 cm proximal the horizontal plane of the femoral head center. The easy-to-penetrate sites were the junction of the acetabular posterior wall and the sciatic branch，the middle part of the acetabulum, and below 1.0 cm of the highest point of greater sciatic notch. The tangential position of three lateral faces of the “triangular prism” were ilium oblique position 10°, the ilium oblique position 60°, and obturator oblique position 60°.  Conclusion The entrance point of the retrograde screw is located on the midline of the ischial tuberosity, which is 1.3 cm from farthest end of the ischial tuberosity and the direction is forward inclination about 15°, external inclination about 10°. The acetabular posterior column fracture that below 4.0 cm proximal plane to the femoral head center can be fixed by the retrograde screw. References | Related Articles | Metrics

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Percutaneous Magic screw fixation for acetabular posterior column fracture: an imaging anatomical study#br# CHEN Jian-fei GAO Shi-chang YU Ke-quan 2020, 51 (1):  86-92.  doi: 10.16098/j.issn.0529-1356.2020.01.015 Abstract ( )   PDF (3032KB) ( )   Objective To investigate the entry point, direction and fixation range of Magic screw for acetabular posterior column fracture.   Methods The 3D models of 100 pelvis were reconstructed by Mimics 19.0 software based on CT data, and the virtual magic screw were placed in acetabular posterior column, then the screw’s entry point were determined, and the direction, length, diameter and safe range were measured respectively. Acetabular posterior column with magic screw was osteotomy modeling by Mimics 19.0 and imported the Geomagic Wrap 2017 software, after that anatomical parameters of the screw path were measured.  Results The entry point of Magic screw on bone surface was located at (33.37±5.53)mm of the anterior inferior iliac spine moving posteriorly, and (13.40±3.70)mm of the apex of the upper margin of acetabular moving celphalad in male, which were (33.97±5.46)mm and (9.01±3.86)mm in female. The posterior inclination angle, interior inclination angle of magic screw and the angle between screw and iliac wing were (57.40±6.57)°, (52.09±5.65)°and (15.21±3.42)°in male, which were (55.64±8.01)°，(51.55±5.58)°and (9.85±3.68)°in female. The maximum diameter of male screw was (6.97±0.98) mm, which was (6.39±0.85) mm in female. The length of male screw was (76.73±9.20) mm, which was (63.64±8.37) mm in female. The safe range of posterior inclination and interior inclination of were (7.19±3.30)°and (9.41±3.95)°in male screw with 5.5 mm diameter, which were (8.37±2.82)°and (10.32±3.93)°in female screw with 5.2 mm diameter. In the direction of the screw, the length range of the posterior column which is fixed by screw was (56.87±7.60) mm，and the proportion of fractures fixed with screws which were located on the top of the acetabular was 20/50 in male, which were (41.71±7.97) mmand 8/50 in female.  Conclusion Percutaneous Magic screw is a minimally invasive treatment for acetabular posterior column fracture, which is difficult to operate. The screw can fix the fracture which is located at the middle and upper part of the acetabular posterior column. References | Related Articles | Metrics

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Vascular anatomy and clinical application of latissimus dorsi myocutaneous flap XIA You-chen LI Bi CHEN Xiao-xun YANG Xin MA Jian-xun ZOU Li-jun WANG Jian-wei ZHANG Wei-guang 2020, 51 (1):  93-97.  doi: 10.16098/j.issn.0529-1356.2020.01.016 Abstract ( )   PDF (1668KB) ( )   Objective To explore the relationship between latissimus dorsi myocutaneous flap and blood supply, so as to provide a scientific basis for the re-division, transposition and transplantation of latissimus dorsi myocutaneous flap.  Methods The latissimus dorsi muscle of 48 cadavers were observed by anatomy and angiography. The clinical applications of latissimus dorsi myocutaneous flap in 31 cases were reviewed.  Results The latissimus dorsi myocutaneous flap had many sources of blood supply. The main thoracodorsal artery was distributed in the upper and outer latissimus dorsi muscle. The medial and lateral branches were separated steadily with their respective distribution areas. The inner and lower parts of latissimus dorsi muscle were supplied by intercostal and lumbar arteries. The anastomosis between them and the branches of thoracodorsal artery was obvious according to X-ray angiography. The caliber was between 320-550 μm. The blood supply of the skin superficial to the latissimus dorsi muscle was from the myocutaneous artery. But the anastomosis between the perforating branches was sparse and the caliber was small near the inner and lower parts. Myocutaneous flaps were applied for wound repair, breast reconstruction and leg defect repair after mass excision in 31 cases. Thirty cases of myocutaneous flaps survived completely post-operation. One case of myocutaneous flap had early signs of partial blood flow disturbance. After treatment, blood flow gradually improved and wound healing delayed.  Conclusion The latissimus dorsi myocutaneous flap can be divided into 3 parts according to their arterial branches and anastomotic characteristics, which can provide the vascular anatomical basis for whole or partial separation, transposition or transplantation and preservation of muscle function.  Related Articles | Metrics

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Anatomical basis in the treatment of sinus tarsi syndrome by local steroid injection LIAO Li-qing LI Hong LI Yi-kai 2020, 51 (1):  98-102.  doi: 10.16098/j.issn.0529-1356.2020.01.017 Abstract ( )   PDF (1594KB) ( )   Objective To provide anatomical basis for the treatment of sinus tarsi syndrome by developing the anatomical structure of sinus tarsi.  Methods Thirty formalin fixed lower limb specimens were dissected. The anatomic structures of the sinus tarsi, neurovascular tissue, ligament and adipose tissue were observed and measured. Morphologies of the sinus tarsi and canal of tarsus in intact dry calcaneus and talus of 136 Chinese adult specimens were investigated and measured.  Results The artery of the sinus tarsi and its periphery issued from the dorsalis pedis artery, anterior medial malleolar artery, perforating peroneal artery and lateral tarsal artery. The laterosuperior point of cuboidal articular surface of calcaneus was the obvious osseous marks of the lateral surface of sinus tarsi. The length canal of tarsus, sinus tarsi  and the calcaneal sulcus were (14.85±2.08)(8.33-22.00)mm, (26.38±3.05) (16.54-35.79)mm and (41.25±3.52)(31.06-53.92)mm respectively. There was a angle between two axes of calcaneus and tarsal canal, which was (49.37±4.76)(39.18-56.37)°. Conclusion During the treatment of sinus tarsi syndrome by the local steroid injection, the laterosuperior point of cuboidal articular surface of calcaneus should be palpated firstly, which is the point of inserting the needle. Then, the injection needle is towards posteromedial at 49°, and advance 2.5 mm to touch the sinus tarsi. References | Related Articles | Metrics

Neurobiology

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Effect of silencing beclin1 gene on apoptosis of HT22 cells after oxygen and glucose deprivation/reoxygenation ZHANG Yi JIN Xiao-fei ZHOU Xiao-hong DONG Xian-hui ZHANG Ying YU Wen-tao CHENG Yuan GAO Wei-juan 2020, 51 (1):  3-8.  doi: 10.16098/j.issn.0529-1356.2020.01.001 Abstract ( )   PDF (3032KB) ( )   Objective To investigate the effect of Beclin1 gene on apoptosis of HT22 of mouse hippocampus neuron treated with oxygen and glucose deprivation/reoxygenation(OGD/R).  Methods HT22 cells in logarithmic growth phase were randomly divided into 4 groups: normal, model, beclin1-/- and control. All groups were reoxygenated after 6 hours of oxygen and glucose deprivation except for normal group. The beclin1 interference sequence was designed for mouse cDNA sequence using RNAi technology, and was transfected into HT22 cells by liposome Lipo2000. The transfection efficiency was observed under fluorescence microscope and Western blotting after 48 hours of transfection. Cell viability was detected by CCK-8 method. Cell damage was detected by lactate dehydrogenase(LDH) method , Bax and Bcl-2 were detected by immunofluorescence staining and the expression of LC3, P62 and Caspase-3 were tested by Western blotting after 24 hours of reoxygenation. SPSS 19.0 statistical software was used for data analysis.  Results Compared with the normal group, the cell viability and P62 expression decreased significantly (P<0.01), the LDH leakage rate, LC3Ⅱ/LC3Ⅰ，Caspase-3 expression and Bax/Bcl-2 increased significantly in model group (P<0.01). Compared with the model group, the cell viability and LC3Ⅱ/LC3Ⅰ decreased significantly (P<0.01), the LDH leakage rate, Bax/Bcl-2，P62 and Caspase-3 expression increased significantly in beclin1-/- group (P<0.01). There was no difference between the control group and the model group.  Conclusion Silencing beclin1 inhibites autophagy, which aggravates the damage of OGD/R HT22 cells and further increases apoptosis. References | Related Articles | Metrics

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Role of hippocampal sortilin in streptozocin inducing diabetic cognitive impairment mice LUO Dan YANG Hui LUO Shi-shi NIU Lei LI Wei SONG Guang-ming XIE Liang-zhe XU Yang CAO Wen-yu HE Jie 2020, 51 (1):  9-14.  doi: 10.16098/j.issn.0529-1356.2020.01.002 Abstract ( )   PDF (1676KB) ( )   Objective To investigate the role of hippocampal sortilin in streptozotocin (STZ) inducing diabetic cognitive impairment in mice. Methods Twenty four adult male ICR mice were randomly divided into control group (NS group) and experimental group (STZ group). The animal model of diabetic cognitive impairment was induced by STZ treatment. The blood glucose was measured by blood glucose meter every other week. The cognitive function was determined by novelty object recognition test 8 weeks after STZ injection. The expressions of sortilin in hippocampus of mice were determined by immunohistochemistry. The expressions of sortilin and brain-derived neurotrophic factor (BDNF）protein in hippocampus of mice were determined by Western blotting. The expressions of sortilin and BDNF mRNA in hippocampus of mice were determined by Real-time PCR. Results STZ treated mice showed significantly increased blood glucose compared with  the NS group (P<0.01). Compared with the NS group, the STZ group showed significantly reduced discrimination index in the novelty object recognition test (P<0.05). The result of immunohistochemistry assay showed that the positive score of NS group was significantly higher than STZ group (P<0.05).The results of Western blotting and Real-time PCR showed that the expression level of sortilin protein (P<0.01) and mRNA (P<0.05) in the hippocampus of STZ group decreased significantly compared with the NS group. The results of Western blotting and Real-time PCR showed that the expression level of BDNF protein (P<0.05) and mRNA (P<0.01) in the hippocampus of STZ group decreased significantly compared with the NS group.Conclusion The STZ-induced cognitive impairment in mice may be related to the down-regulation of sortilin in the hippocampus. References | Related Articles | Metrics

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Effect of Notch signaling pathway on nerve regeneration after cerebral ischemia in rats with neural stem cells transplantation#br# LIU Zong-xiu ZHANG Zi-wei CHENG Yuan JIN Xiao-fei ZHOU Xiao-hong GAO Wei-juan 2020, 51 (1):  15-20.  doi: 10.16098/j.issn.0529-1356.2020.01.003 Abstract ( )   PDF (2654KB) ( )   Objective To investigate the mechanism of inhibiting the Notch signaling pathway in promoting neural regeneration after neural stem cells (NSCs) transplantation in rats with cerebral ischemia.  Methods The model of focal cerebral ischemia was established by middle cerebral artery occlusion (MCAO) method . NSCs were cultured in vitro and transplanted into the striatum ischemic area. In the experiment, 40 SD rats were divided into sham operation group, model group, transplantation group (transplanted neural stem cells), and N-［N-(3,5-difluorohenacetyl)-L-alanyl］-S-phenylglycinet-butyl ester（DAPT）+ transplantation group. The degree of neuronal damage in each group was observed by HE staining. The expressions of Notch1, Hes1 and Hes5 in the brain tissue of each group were detected by immunohistochemistry and Western blotting.  Results Compared with the sham operation group, the neuron injury in the model group was severe, and nuclear pyknosis and nuclear lysis were observed. The Notch1, Hes1 and Hes5 positive cells increased significantly, and the expression of Notch1, Hes1 and Hes5 proteins were significantly up-regulated (P<0.05). Compared with the model group, the neuronal damage in the transplantation group and the DAPT+ transplantation group were all relieved, and the Notch1, Hes1 and Hes5 positive cells were partially expressed, and the expression of each protein decreased (P<0.05). The DAPT+ transplantation group neurons were compared. The damage was obviously restored, and the expression of protein-positive cells and protein further decreased (P<0.05).  Conclusion Inhibition of Notch signaling pathway can promote nerve regeneration after neural stem cell transplantation in rats with cerebral ischemia. The mechanism is mainly related to down-regulation of Notch1, Hes1 and Hes5 expression. References | Related Articles | Metrics

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Combining effects of aspirin and nimodipine on prognosis of cerebral ischemia reperfusion#br# LIU Qian HU Ya-guang CHAI Er-qing 2020, 51 (1):  21-25.  doi: 10.16098/j.issn.0529-1356.2020.01.004 Abstract ( )   PDF (708KB) ( )    Objective To investigate the effects of the combination of aspirin and nimodipine preconditioning on the prognosis of cerebral ischemia-reperfusion.  Methods Eighty healthy male SD rats were randomly divided into sham group, model group, aspirin preconditioning group and aspirin + nimodipine preconditioning group, with 20 rats in each group. The model of cerebral ischemia reperfusion was established. The rats in each group were given intragastric administration for 5 days before the model was established, and the drug was administered daily for 5 days. Sham group and model group were given normal saline; Aspirin preconditioning group was given 50 mg/kg aspirin; Aspirin + nimodipine preconditioning group was given 50 mg/kg aspirin and 10 mg/kg nimodipine. After 2 hours ischemia and 24 hours reperfusion, the animals were neurologically assessed, and then the volume of cerebral infarction was measured by TTC staining. The contents of superoxide dismutase (SOD), malondialdehyde (MDA), thromboxane B2, 6-keto-prostaglandin-1α in brain tissue were determined by ELISA. The mRNA expression of Notch1, Jagged1 and Hes1 in the brain tissues were detected by Real-time PCR, and the expressions of Jagged1 and Hes1, a downstream substance in Notch signaling pathway, were detected by Western blotting.  Results The neurological deficit score of the aspirin+nimodipine pretreatment group was significantly lower than model group (P<0.05), and the cerebral infarction volume was significantly smaller than other groups. The SOD and 6-keto-prostaglandin 1 in the aspirin pretreatment group and the aspirin plus nimodipine pretreatment group were significantly higher than those in the model group, and the expressions of MDA, thromboxane B2 and thromboxane B2/6-keto-prostaglandin 1 were low. In the model group, the changes in the aspirin + nimodipine pretreatment group were more significant (P<0.05). The expression levels of Notch1, Jagged1 and Hes1 mRNA and protein in the aspirin pretreatment group and aspirin+nimodipine pretreatment group were significantly lower than those in the model group (P<0.05), and the expression level of aspirin+nimodipine pretreatment group was lower than that of aspirin pretreatment group (P<0.05).  Conclusion The protective effect of aspirin plus nimodipine is superior to aspirin alone, which can significantly improve cerebral ischemia-reperfusion injury in rats, which may be exerted through influencing notch signaling pathway to achieve brain tissue protection. References | Related Articles | Metrics

Cancer Biology

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Enhancing effect of Lupeol on migration and invasion abilities of ST3GalⅢ-silenced MDA-MB-231 cells#br#  #br# WANG Ming CUI Hong-xia LIU Ji-cheng 2020, 51 (1):  26-31.  doi: 10.16098/j.issn.0529-1356.2020.01.005 Abstract ( )   PDF (2222KB) ( )   Objective In this study, we expored the enhancing effect of Lupeol on migration and invasion abilities of ST3GalⅢ-silenced MDA-MB-231 cells.  Methods Human breast cancer cell line ST3GalⅢ -silenced MDA-MB-231 was cultured in vitro. The cell adhesion, Transwell and woud healing test were utilized to test the effect of Lupeol on ST3GalⅢ-silenced MDA-MB-231 cells. The expression of matrix metalloproteinase(MMP)-2, -9, phosphatidylinositol 3-kinase/protein kinase B/nuclear factor κB(PI3K/Akt/NF-κB) signaling pathway were examined by Western blotting.  Results No significant influence of the decreased expression of ST3GalⅢ and Lupeol（5μmol/L）on proliferation and apoptosis of MDA-MB-231 cells were found. Lupeol inhibited the migration and invasion of ST3GalⅢ-silenced MDA-MB-231 cells in vitro(P<0.05). Furthermore, the expression of NF-κB p65, p-Akt, Akt, p85, MMP-2 and MMP-9 levels were significantly down-regulated.  Conclusion These observations suggest that Lupeol may inhibit the abilities of migration and invasion of ST3GalⅢ-silenced MDA-MB-231 cells in vitro by inhibiting the protein expression of MMP-2, -9 and effect the signaling pathway of PI3K/Akt/NF-κB. References | Related Articles | Metrics

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Anti-apoptotic effect of human acute myeloid leukemia cell line HL-60 stimulated by interleukin-6 from bone marrow stromal cells HS-5 CHEN Ya-lin BAI Tian-ge LI Yao-yao WANG Wen-qiang WEI Hong 2020, 51 (1):  32-39.  doi: 10.16098/j.issn.0529-1356.2020.01.006 Abstract ( )   PDF (3990KB) ( )   Objective To investigate the effect of interleukin(IL)-6 secreted by bone marrow stromal cells HS-5 on activity and apoptosis of human acute myeloid leukemia(AML) cells HL-60 and its possible mechanism.  Methods HL-60 cells and HS-5 cells were cultured in vitro, and the co-culture system was established. Scanning electron microscope, ELISA, cell counting kit-8(CCK-8), AnnexinV-FITC/PI, double-staining flow cytometry, Real-time PCR and Western blotting techniques were used to detect the changes of viability and apoptosis of HL-60 cells, respectively. HL-60 cells from different groups were inoculated into BALB/c nude mice to observe and record tumorigenesis.  Results IL-6 secreted by bone marrow stromal cells HS-5 could enhance the viability and inhibit the apoptosis of HL-60 cells, and down-regulate the expression of pro-apoptotic gene Bax and and up-regulate the expression of anti-apoptotic gene Bcl-2 in HL-60 cells. HL-60 cells in co-culture group had the strongest tumorigenicity in BALB/c nude mice, while HL-60 cells alone had the weakest tumorigenicity.  Conclusion Part of the mechanism by which bone marrow stromal cells HS-5 promote the proliferation of HL-60 cells and inhibit their apoptosis may be through the secretion of IL-6. References | Related Articles | Metrics

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Effect of lentiviral vector mediated RNA binding protein quaking-5 on the proliferation and apoptosis of lung adenocarcinoma cell line H1299 FANG Ke-xin WANG Xin-chen HONG Kai-ting ZHU Wang-yu 2020, 51 (1):  40-46.  doi: 10.16098/j.issn.0529-1356.2020.01.007 Abstract ( )   PDF (3456KB) ( )    To investigate the proliferation and apoptosis of lung adenocarcinoma cells line H1299 by the lentiviral vector mediated RNA binding protein quaking-5 (QKI-5).  Methods GV358 (up-regulation) and GV248 (down-regulation) vectors were used to construct the lentiviral QKI-5 up-regulation vector and down-regulation vector, respectively. The vectors were transfected into 293T cells for lentiviral packaging and viral titer were then determined. Gene sequencing was performed to screen the sequence of vectors. Then Real-time PCR was used to evaluate the expression of QKI-5 mRNA and the proliferation of H1299 cells was examined by colony forming assay after transfection. The apoptosis of H1299 cells was determined by the detection of the expression membrane protein Ⅴ (annexin Ⅴ) and propyl iodide ingot (PI) by flow cytometry. Pro-apoptotic protein Caspae-3 and Caspase-8 were evaluated by Western blotting.  Results QKI-5 up-regulation and down-regulation lentiviral vectors were constructed successfully. Compared with the controls, the expression of QKI-5 mRNA of H1299 cells was up-regulated, the cell colony formation was decreased, and the early apoptosis of H1299 cells was increased with the over-expression of Caspase-8 after transfected with up-regulated vector, whereas transfecting with QKI-5 down-regulated vector had opposite effect.  Conclusion Lenviral vector mediated QKI-5 could inhibit proliferation and promote apoptosis of lung adenocarcinoma cells through Caspase-8. References | Related Articles | Metrics

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Gambogic acid suppresses invasion in human gastric carcinoma SGC-7901 cells by modulating inhibitor of nuclear factor kappa-B kinase α/p65 signaling pathway in vitro#br# TONG Lei WANG Shi-guang YANG Zhan-feng YUE Li-xiao ZHANG Ling-ling WANG Peng 2020, 51 (1):  47-50.  doi: 10.16098/j.issn.0529-1356.2020.01.008 Abstract ( )   PDF (2392KB) ( )   Objective To investigate the effect of gambogic acid (GA) on invasion in human gastric carcinoma SGC-7901 cells and its possible mechanism.  Methods Cell counting kit-8(CCK-8) assay was performed to detect the effects of GA, inhibitor of nuclear factor kappa-B kinase(IKK)16 and 5-fluorouracil (5-FU) on cell activity of GES-1 and SGC-7901 cells. Cell invasion was assessed with Transwell invasion assay. Western blotting was used to analyze the protein levels of vimentin, matrix metalloproteinase 2 (MMP-2) and MMP-9 and protein phosphorylation of IKKα and p65.  Results The cell activity was significantly decreased in SGC-7901 cells treated with GA in a dose-dependent manner with a half inhibiton concentration(IC50) value of 1.89 μmol/L. But GA had no significant influence on cell viability of GES-1 cells. Meanwhile, 5-FU reduced the cell activity of GES-1 and SGC-7901 cells with IC50 values of 7.36 μmol/L and 199.57 μmol/L respectively. Low-dose GA and IKK16 impaired separately the ability of invasion in SGC-7901 cells, and down-regulated the protein levels of MMP-2, MMP-9 and vimentin, and inhibited phosphorylation of IKKα and p65, while a stronger inhibition was showed when the combination of GA and IKK16 was used.  Conclusion Low-dose GA might inhibit invasion of SGC-7901 cells via IKKα/p65 signaling pathway. References | Related Articles | Metrics

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Screening and survival analysis of differentially expressed genes in papillary thyroid carcinoma#br# #br# LIU Zhong-jiao DOU Long-tao ZHUANG Zhen LIU Ling ZHOU Yong-wei LIU Man LI Jing ZHANG Hong-mei 2020, 51 (1):  51-57.  doi: 10.16098/j.issn.0529-1356.2020.01.009 Abstract ( )   PDF (4988KB) ( )   Objective Bioinformatics method  was used to analyze gene expression microarrays of papillary thyroid cancer (PTC) and adjacent tissues. The key genes of PTC and their signal pathways were screened to understand their carcinogenic mechanisms.  Methods Data on PTC and paracancerous tissue samples were obtained from seven GSE series on two sequencing platforms in the Gene Expression Omnibus Database(GEO).Firstly, the differential genes of the two sequencing platform samples were screened by R language. Then biological function, signal pathway analysis and protein-protein interaction analysis were performed on differential genes by Metascape and STRING. Finally, the key gene were selcected by Cytoscape 3.5.1 software.  Results A total of 302 differential genes were obtained from the intersection of the two sequencing platform samples, of which 149 genes were up-regulated and 153 genes were down-regulated. Using the Cytoscape 3.5.1 software to screen out 15 key genes, 12 of them are involved in the extracellular matrix receptor interaction signal pathway. Survival analysis of 15 key genes was performed using the UALCAN database, and the changes in the expression levels of 4 genes were closely related to the survival time of patients.  Conclusion This study uses bioinformatics technology to analyze the data from seven PTC gene chips, making up for the inconsistency of small sample result  and improving the reliability and stability of the result . In addition, 15 key genes are screened out and found that the matrice extracellulaire receptor interactions pathway plays an important role in the development of thyroid cancer. The result  of this experiment provides guidance for the further study  of PTC molecular mechanism, diagnosis and screening of prognostic molecular markers. References | Related Articles | Metrics

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Mechanism of microRAN-145 on proliferation and apoptosis of breast cancer cell line MCF-7#br# QI Yu-lin HAN Hui-fang ZHANG Hai-xin 2020, 51 (1):  58-61.  doi: 10.16098/j.issn.0529-1356.2020.01.010 Abstract ( )   PDF (1081KB) ( )   Objective To explore the mechanism of microRNA-145 (miR-145) involved in proliferation and apoptosis of breast cancer MCF-7 cells.  Methods The immortalized breast cancer cell line MCF-7 cells were cultured in vitro and transfected with miR-145. Real-time PCR and Western blotting were used to detect mRNA and protein levels, respectively. The proliferation level of each group was detected by cell counting kit-8 (CCK-8) method , and the apoptosis level of MCF-7 cells in different treatment groups was detected by flow cytometer.  Results Real-time PCR result showed that miR-145 did not affect Caspase-3, proliferating cell nuclear antigen (PCNA) and B cell lymphoma factor 2 (Bcl-2) mRNA levels in MCF-7 cells. Western blotting analysis showed that compared with the control group, transfection of miR-145 for 96 hours significantly increased the expression of Caspase-3 and inhibited the expression of PCNA and Bcl-2. The result of CCK-8 assay showed that the proliferation rate of MCF-7 cells was decreased after overexpression of miR-145 for 72 hours and 96 hours (P<0.05). The result  of flow cytometer showed that the apoptosis rate of MCF-7 cells in overexpression group was significantly higher than that in the control group (P<0.05).  Conclusion MiR-145 can inhibit the proliferation and promote the apoptosis of MCF-7 cells by down-regulating PCNA and Bcl-2 and up-regulating the expression of Caspase-3, which may be a new target for breast cancer treatment. References | Related Articles | Metrics

Bioengineering

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Preparation and histological evaluation of human decellularized adipose tissue ZHAO Yu- FAN Jun BAI Shu-ling 2020, 51 (1):  128-131.  doi: 10.16098/j.issn.0529-1356.2020.01.022 Abstract ( )   PDF (2850KB) ( )   Objective To decellularize adipose tissue derived from human liposuction, evaluate the histological composition of human decellularized adipose tissue, and explore the possibility for it as tissue engineering scaffold materials.  Methods The adipose tissue derived from five human liposuction was decellularized by a series of physical chemistry and enzymology method . The general morphology of the decellularization process was observed. HE staining, Masson staining, Gomori staining, oil red O staining and DAPI staining were performed and the content of DNA, collagens and glycosaminoglycans remaining in the decellularized adipose tissue were quantitatively detected.  Results The decellularized adipose tissue was some kind of milky, soft, variable insoluble material. HE staining and DAPI staining showed no residual cell components. Masson staining showed the presence of collagen fibers. Gomori staining showed the presence of reticular fibers, and oil red O staining showed no visible lipid residues. The residual DNA content in the tissue is very tiny and the collagen and glycosaminoglycan contents were retained.  Conclusion The adipose tissue derived from the body’s liposuction can be decellularized by physical, chemical, and enzymatic method . The cells and lipid in the decellularized adipose tissue can be effectively removed, and the components of the extracellular matrix of the adipose tissues are mostly retained. References | Related Articles | Metrics