1.Fabrication of the composite scaffolds for bone regeneration and verification of their value in muscle pouch osteogenic activity in rats
Ming ZHENG ; Qiang ZHOU ; Jixing YE ; Zongxin LI ; Yaopeng ZHANG ; Xiang YAO ; Xueping WEN ; Nan WANG ; Dianming JIANG
Chinese Journal of Trauma 2024;40(7):635-646
Objective:To fabricate the composite scaffolds for bone regeneration with silk fibroin (SF), bacterial cellulose nanofibers (BCNR) and hydroxyapatite (HAp) and evaluate their osteogenic activity.Methods:HAp particles, BCNR and bone morphogenetic protein-2 (BMP2) were added into SF aqueous solution in turn, poured into molds of different sizes after being mixed evenly and processed at -25 ℃ for 24 hours to obtain frozen molds, and the composite scaffolds were frozen-dried by freezing-drying machine. The composite scaffolds with different mass ratios of SF and BCNR were divided into groups A (2∶1), B (4∶1) and C (6∶1), and the inactive composite scaffolds without BMP2 fell into group D. The surface morphology and pore structure of the scaffolds were detected by scanning electron microscopy. The porosity of the scaffolds was measured by mercury intrusion porosimeter. The stress-strain curve was obtained by using the universal material testing machine to compress the scaffolds, with which their compressive strength and Young′s modulus were analyzed. Immortalized mouse embryonic fibroblasts (iMEF) were inoculated on the composite scaffolds of group A, B, C and D. At 4 and 8 days after cell inoculation, the proportion of alive and dead cells in each group was detected by cell survival/death staining; the cell counting kit-8 (CCK-8) was used to detect cell proliferation activity in each group; the positive staining cells were detected in each group by alkaline phosphatase (ALP) staining; the ALP activity was observed in each group with ALP activity detection. A total of 15 female SD rats were selected to establish osteogenesis models with ectopic muscle bag. The composite scaffolds implanted with different SF/BCNR mass ratios and the inactive composite scaffolds without BMP2 fell into group A′ (2∶1), B′ (4∶1), C′ (6∶1) and D′ respectively, and a sham operation group was set at the same time, with 3 rats in each groups. In the sham operation group, the muscle bag and skin were sutured without scaffold implantation after the incision of skin, the blunt separation of the quadriceps muscle, and the formation of muscle bag in the muscle. In the other four groups, the corresponding scaffolds were implanted in the muscle bag and the muscle bag and skin were sutured. X-ray examination was performed at 2 and 4 weeks after operation to observe the osteogenesis in each group. At 4 weeks after operation, the implanted scaffolds and tissue complexes were collected by pathological tissue sectioning, HE staining and Masson staining, and for observing the osteogenesis by in each group. Immunohistochemical staining was also performed on the tissue sections to observe the expression of osteogenic markers type I collagen (COL1) and osteopontin (OPN) in each group.Results:Scanning electron microscopy showed that the lamellar and micropore structures of group B were more regular and uniform than those of groups A and C. The porosity rate analysis showed that the porosity rates of groups B and C were (89.752±1.866)% and (84.257±1.013)% respectively, higher than that of group A [(81.171±1.268)%] ( P<0.05 or 0.01), with the porosity rate of group C lower than that of group B ( P<0.01). The mechanical property test showed that the compressive strengths of groups B and C were (0.373±0.009)MPa and (0.403±0.017)MPa respectively, higher than that of group A [(0.044±0.003)MPa] ( P<0.01), and the Young′s moduli of groups B and C were (7.413±0.094)MPa and (9.515±0.615)MPa respectively, higher than that of group A [(1.881±0.036)MPa] ( P<0.01), with the compressive strength and Young′s modulus of group C higher than those of group B ( P<0.05 or 0.01). The cell survival/death staining showed that the number of dead cells of group B was significantly smaller than that of groups A, C and D at 4 days after cell inoculation, and that group B had the most living cells and the fewest dead cells at 8 days after cell inoculation. The results of CCK-8 experiment showed that at 4 days after cell inoculation, the cell proliferation activity of groups A and B was 0.474±0.009 and 0.545±0.018 respectively, higher than 0.394±0.016 of group D ( P<0.01); the cell proliferation activity of group C was 0.419±0.005, with no significant difference from that of group D ( P>0.05), while the cell proliferation activity of groups A and C were both lower than that of group B ( P<0.01). At 8 days after cell inoculation, the cell proliferation activity of group B was 1.290±0.021, higher than 1.047±0.011 of group D ( P<0.01); the cell proliferation activity of group C was 0.794±0.032, lower than that of group D ( P<0.01); the cell proliferation activity of group A was 1.086±0.020, with no significant difference from that of group D ( P>0.05); the cell proliferation activity of groups A and C was lower than that of group B ( P<0.01). At 4 and 8 days after cell inoculation, ALP staining showed that more positive cells were found in groups A, B and C when compared with group D, and more positive cells were found in group B than in groups A and C. At 4 days after cell inoculation, the ALP activity detection showed that the ALP activity of groups A, B and C was 1.399±0.071, 1.934±0.011 and 1.565±0.034 respectively, higher than 0.082±0.003 of group D ( P<0.01), while the ALP activity of groups A and C was lower than that of group B ( P<0.01). At 8 days after cell inoculation, the cell activity of groups A, B and C was 2.602±0.055, 3.216±0.092 and 2.145±0.170 respectively, higher than 0.101±0.001 of group D ( P<0.01), while the ALP activity of groups A and C was lower than that of group B ( P<0.01). X-ray examination results showed that at 2 weeks after operation, no obvious osteogenesis was observed in the sham operation group, group D′, A′ and C′, while it was observed in group B′. At 4 weeks after operation, obvious osteogenesis was observed in group A′, B′ and C′, with significantly more osteogenesis in group B′ than in the other two groups, while there was no obvious osteogenesis in the sham operation group and group D′. At 4 weeks after operation, the HE staining and Masson staining showed that a large number of uniformly distributed new bone tissue was formed in group B′, while only a small amount of new bone tissue was found locally in groups A′ and C′, and only part of new tissue was found to grow in group D′ with no obvious new bone tissue observed. The maturity of new bone tissue formed in group B′ was higher than that in group A′ and C′. Immunohistochemical staining showed more COL1 and OPN positive staining in group B′ when compared with groups A′ and C′. The expression intensity analysis of COL1 and OPN showed that in groups A′, B′ and C′, the expression intensity of COL1 was 2.822±0.384, 22.810±2.435 and 12.480±0.912 respectively and the expression intensity of OPN was 1.545±0.081, 5.374±0.121 and 2.246±0.116 respectively, with higher expression intensity of COL1 and OPN in groups B′ and C′ than that in group A′ ( P<0.01) and lower expression intensity of COL1 and OPN in group C′ than that in B′ group ( P<0.01). Conclusions:The composite scaffold for bone regeneration is successfully fabricated with SF, BCNR and HAp. The composite scaffold with a mass ratio of SF to BCNR of 4∶1 has uniform pore structure, high porosity, good mechanical properties and biocompatibility, excellent pro-osteogenic properties in vitro, as well as excellent osteo-inductivity and osteo-conductivity.
2.Effect of SMAC Gene on Sensitivity of Lung Adenocarcinoma Cells to Paclitaxel and Cell Viability Based on caspase-3/Bcl-2/Bax Signaling Pathway
Kang CHEN ; Ying CHEN ; Zongxin NIU ; Li KANG
Cancer Research on Prevention and Treatment 2023;50(4):357-363
Objective To investigate the effect of the SMAC gene on paclitaxel sensitivity and cellular activity in lung adenocarcinoma cells based on the caspase-3/Bcl-2/Bax signaling pathway. Methods A paclitaxel-resistant cell line A549/Taxol was established for lung adenocarcinoma, and the cells were divided into four following groups: pcDNA-NC (transfected with pcDNA-NC blank vector), pcDNA-SMAC (transfected with pcDNA-SMAC vector), siRNA-NC (transfected with siRNA-NC empty virus vector), and siRNA-SMAC groups (transfected with siRNA-SMAC lentiviral vector). The SMAC mRNA expression in cells was detected by qRT-PCR; cell sensitivity was detected by MTT; cell proliferation ability was detected by cloning assay; cell invasion ability was detected by Transwell; apoptosis ability was detected by flow cytometry assay; and caspase-3, Bcl-2 and Bax protein expression in cells were detected by Western blot analysis. Results The SMAC mRNA expression was significantly lower in A549 cells compared with BEAS-2B cells (
3.Standard on classification of Traditional Chinese Medicine ancient books
Huamin ZHANG ; Weina ZHANG ; Hongtao LI ; Yanping WANG ; Xuejie HAN ; Yuqi LIU ; Lin TONG ; Bing LI ; Sihong LIU ; Hongyan CHEN ; Zongxin WANG ; Hao QIU
International Journal of Traditional Chinese Medicine 2023;45(1):1-7
The standardization of classification methods of Traditional Chinese Medicine(TCM) ancient books can provide a clear and reliable reference for all kinds of TCM ancient books collection units, which can also promote the sharing and utilization of TCM ancient books. We studied and investigated the classification methods of TCM ancient books in past dynasties. The standard on classification of TCM ancient books was formulated by compared with the classification table of Zhongguo Zhongyi Guji Zongmu, and referred to the classification table of Zhonghua Guji Zongmu. This standard specified three-level categories and classification principles of TCM ancient books, and mainly composed of basic categories, three-level category table, classification principles and examples, and instructions for use.
4.A preliminary clinical study comparing percutaneous transhepatic choledochoscopy lithotomy with laparoscopic choledocholithotomy for choledocholithiasis
Tianling FANG ; Zongxin WANG ; Anzhong LIU ; Yanmin LIU ; Canhua ZHU ; Jutao FENG ; Jun LI
Chinese Journal of Hepatobiliary Surgery 2020;26(11):854-857
Objective:To compare the clinical efficacy of percutaneous transhepatic choledochoscope lithotomy (PTCSL) with laparoscopic choledocholithotomy (LD) in treatment of choledocholithiasis.Methods:Data of 132 patients with choledocholithiasis treated at the First Affiliated Hospital of Guangzhou Medical University from July 2012 to December 2018 were retrospectively analyzed. There were 75 males and 57 females, with an average age of 62.7 years. For 76 patients underwent PTCSL (the PTCSL group) and 56 underwent LD (the LD group). The data of the patients the success rate of lithotomy, stone residual rate, operation time, postoperative complications and stone recurrence, chronic cholangitis, and acute cholangitis 1 month after operation were compared between the two groups.Results:The ratio of upper abdominal operation history and biliary tract infection in the PTCSL group was higher than that in the LD group, and the difference was statistically significant (both P<0.05). In the PTCSL group, the calculi were successfully removed in 64 patients in one treatment session, while residual calculi were removed through subsequent sinus choledochoscopy in 9 patients. In the remaining 3 patients, the residual calculi were removed with LD or laparotomy operations. Postoperative complications occurred in 14 patients (19.2%, 14/73). In the LD group, the calculi were successfully removed in one session in 46 patients while in 8 patients the residual calculi were removed by choledochoscopy (1 patient still had residual calculi after choledochoscopy). The remaining 2 patients underwent open surgery due to anatomical difficulties. Postoperative complications occurred in 11 patients (20.4%, 11/54). There were no significant differences between the two groups in the one-off stone removal rate, postoperative stone residual rate, final stone removal rate and postoperative complication rate (all P>0.05). The operation time of the PTCSL group was (156±60) min, which was significantly shorter than the LD group (203±59) min ( P<0.05). There was no significant difference between the two groups in the incidence of postoperative chronic cholangitis and recurrence rate of calculi (both P>0.05). The incidence of acute cholangitis in the PTSCL group was significantly higher than that in the LD group ( P<0.05). Conclusion:PTCSL was as safe and effective as LD, with fewer complications and faster recovery. It is especially suitable for patients with previous upper abdominal surgery, recurrence of calculi and repeated biliary tract infection.
5.Application of internal iliac artery embolization and presetting abdominal aorta balloon for complicated pelvic frac-tures
Xiaodong YANG ; Han LIU ; Zongxin ZHOU ; Weiyu HAN ; Guang XIA ; Cheng GU ; Tao LI ; Weiqi HUANG ; Qiguang MAI ; Dadi JIN ; Shicai FAN
Chinese Journal of Orthopaedics 2017;37(1):11-16
Objective To evaluate the clinical outcome of bleeding control by preoperative embolization of internal iliac artery with DSA and intra?operative presetting abdominal aorta balloon, combine with the operation techniques of exposure, reduc?tion and internal fixation of pelvic fracture through lateral?rectus approach. Methods From March 2012 to May 2015, 7 patients with type C3 pelvic fractures admitted to our department from March 2012 to May 2015, treated with preoperative embolization of internal iliac artery under digital subtraction angiography 2 h before surgery and presetting abdominal aorta balloon were retrospec?tively reviewed. There were 3 males and 4 females, with an average age of 34 years (range, 16 to 61 years). According to AO classi?fication, all 7 cases belonged to type C3 (3.2:5 cases;C3.3:2 cases), including 5 cases with limb fracture, 2 cases with craniocere?bral trauma, 4 cases with pulmonary contusion, 2 cases with injury of abdominsal organs. Time from injury to operation was 19 days on average (10 to 33 days). Patients received damage control surgery treatment including bleeding control and temporary ex?ternal fixation, and ipsilateral tractions with heavy weight, intensive care and corrections of general situation before operation. The fracture model was manufactured by 3D printing and fracture reduction was simulated on computer preoperatively. Embolization of internal iliac artery was performed in the side of severe displaced sacroiliac joint with DSA 2 hours preoperatively. Reduction was performed to stabilize anterior-posterior pelvic ring and acetabular fractures via the intraoperative lateral?rectus approach. And 2 cases were performed by temporary balloon occlusion of abdominal aorta (≤60 min) for bleeding control in reduction of in the side of sacroiliac joint fractures. Results All the 7 cases had undergone the operations successfully, and the operating time was from 135-320 min with blood loss from 440-3 350 ml. According to Matta radiological evaluation postoperatively, reduction of pelvic fracture was rated as anatomic in 5 cases, satisfactory in 2, without complications. All 7 cases were complicated with lumbosacral plexus injury or lumbosacral trunk injury at different degrees (M0 2 cases, M1 2 cases, M2 2 cases, M3 1 case). According to the BMRC scoring system, 5 cases had well recovered and the other 2 cases had no improvement after three months (M4 2 cases, M5 3 cases). Conclusion Surgical management of pelvic fracture through preoperative internal iliac artery embolization and intra?oper?ative occlusion of abdominal aorta could effective control bleeding and achieve favorable conditions for reduction. Lateral?rectus approach can provide adequate exposure of the anterior and posterior ring, and this approach could also provide excellent visual control of reduction and fixation.
6.Effects of naokang erhao decoction on cognitive ability and hippocampal apoptosis-related proteins in diabetic rats.
Yujing CHEN ; Lin LI ; Zongxin LI ; Xiaobo HUANG ; Li ZHANG ; Wenqiang CHEN
China Journal of Chinese Materia Medica 2011;36(11):1519-1523
OBJECTIVETo investigate the effects of Naokang Erhao decoction on the cognitive ability and the expression of Caspase-3, Bax and Bcl-2 proteins in hippocampus of type 2 diabetic rats.
METHODThe diabetes mellitus (DM) rat model was produced by intraperitoneal injection of streptozotocin and fed with high fat and sucrose diet. The Naokang Erhao-treated rats were intragastrically given different doses of Naokang Erhao, whereas the control and DM model groups were given double distilled water for 4 consecutive weeks. Learning and memory abilities of rats were tested with the Morris water maze. The expression of Caspase-3, Bax and Bcl-2 proteins in hippocampal CA1 region was measured by immunohistochemistry.
RESULTBoth escape latency and swimming distance of type 2 DM rats were significantly prolonged in comparison of those in normal control (P < 0.01), and swimming time in the platform of previous quadrant was significantly shorter in model group (P < 0.01). Furthermore, the expression of Bcl-2 protein was decreased, while Caspase-3 and Bax in the hippocampus were increased compared with the control group (P < 0.01). Four weeks of treatment with Naokang Erhao decoction remarkably improved the learning and memory abilities of DM rats, increased the expression of Bcl-2 and decreased the expression of Caspase-3 and Bax in hippocampal CA1 region of model rats (P < 0.01 or P < 0.05).
CONCLUSIONNaokang Erhao decoction may inhibit apoptosis by increasing the expression of Bcl-2 and reducing the expression of Caspase-3 and Bax in the hippocampus, and this may be one of the mechanisms by which Naokang Erhao decoction improves cognitive ability in DM rats.
Acetophenones ; pharmacology ; Animals ; Apoptosis ; drug effects ; Benzoates ; pharmacology ; Bridged-Ring Compounds ; pharmacology ; CA1 Region, Hippocampal ; drug effects ; metabolism ; Caspase 3 ; drug effects ; metabolism ; Cognition Disorders ; drug therapy ; metabolism ; Diabetic Neuropathies ; drug therapy ; metabolism ; Diterpenes, Abietane ; pharmacology ; Drug Combinations ; Drugs, Chinese Herbal ; pharmacology ; Glucosides ; pharmacology ; Male ; Maze Learning ; drug effects ; Proto-Oncogene Proteins c-bcl-2 ; drug effects ; metabolism ; Pyrazines ; pharmacology ; Rats ; bcl-2-Associated X Protein ; drug effects ; metabolism
7.Anxiety of Cerebral Infarction Patients with Sleep-disordered Breathing and Correlated with Sleep Quality
Ningqun WANG ; Zongxin LI ; Xiaobo HUANG ; Wenqiang CHEN
Chinese Journal of Rehabilitation Theory and Practice 2010;16(5):446-448
ObjectiveTo investigate the anxiety and its effect on the sleep quality in patients with cerebral infarction and sleep-disordered breathing. Methods149 cerebral infarction patients were divided as 95 without sleep-disordered breathing and 54 with sleep-disordered breathing. They were evaluated with Hamilton Anxiety scale (HAMA) and Pittsburgh Sleep Quality Index (PSQI). ResultsPatients with sleep-disordered breathing showed higher HAMA overall score and factor scores of somatic anxiety and psychic anxiety(P<0.001). In the factors of somatic anxiety, the scores of sensing, cardiovascular, gastrointestinal and autonomic nerve system were higher in patients with sleep-disordered breathing than those without sleep-disordered breathing (P<0.05). In the factors of psychic anxiety, the scores of anxious mood, tension, fears, insomnia, depressed mood were higher in patients with sleep-disordered breathing(P<0.05). PSQI overall score and all factor scores were also higher in patients with sleep-disordered breathing(P<0.001). HAMA overall score as well as psychic anxiety and somatic anxiety scores were closely correlated with the PSQI overall score (P<0.001). Somatic anxiety score was correlated with subjective sleep quality, sleep latency, sleep disturbance and daily dysfunction scores (P<0.05). Psychic anxiety score was correlated with all the dimension scores of PSQI (P<0.01). ConclusionCerebral infarction patients with sleep-disordered breathing appeared anxiety and worse sleep quality than patients without sleep-disordered breathing. Anxiety is closely related with sleep quality.
8.Bibliographic Study on Symptomatic Figures of Lip and Tongue (Chunshe Zhenghou Tu)
Zongxin WANG ; Jian QIU ; Rong LIANG ; Hongtao LI ; Jinsheng ZHENG
International Journal of Traditional Chinese Medicine 2009;31(2):157-158
For all the time the book Symptomatic Figures of Lip and Tongue(Chunshe Zhenghou Tu)was described as written by Li Jun of Qing dynasty. We made studies on historical records on Li Jun and the features and contents of this book, and proved that Symptomatic Figures of Lip and Tongue was not written by him, but a Chinese medical book written by Japanese.
9.Clinical Characteristics of Comorbid Anxiety and Depression after Cerebral Infarction
Ningqun WANG ; Zongxin LI ; Xiaobo HUANG ; Wenqiang CHEN
Chinese Journal of Rehabilitation Theory and Practice 2009;15(5):455-457
Objective To determine the clinical characteristics of comorbid anxiety and depression after cerebral infarction. Methods All the patients enrolled were divided into post-stroke comorbid anxiety and depression (PSCAD) group (84 cases) and the control group (121 cases). Their condition of smoke, alcohol intake, education background, carotid plaque and brain blood vessels were investigated, and they were evaluated with Hamilton Anxiety Scale (HAMA), Hamilton Depressive Scale (HAMD), Barthel index, and Mini-Mental State Examination (MMSE). Results There was not significant difference in condition of smoke, alcohol intake, education background, carotid plaque and brain blood vessels between these groups (P>0.05), but were in total scores and all the factors scores of HAMA and HAMD (P<0.05). PSCAD group showed more obvious anxious mood, depressive mood, tension, insomnia, cognitive impairment and autonomic nervous symptoms. Psycho-anxiety, soma-anxiety, despair, guiltiness, decreased interest were also more significant in the PSCAD group. The scores of MMSE and Barthel index were also impaired. Conclusion The cognitive function and daily living ability are worse in the patients with PSCAD. The prominent symptoms includes: anxious mood, depressive mood, tension, insomnia, cognitive impairment, autonomic nervous symptoms, soma-anxiety, despair, guiltiness, decreased interest.
10.Relationship between Carotid Atherosclerosis and Cognitive Impairment or Quality of Life in Patients with Cerebral Infarction
Xiaobo HUANG ; Zongxin LI ; Ningqun WANG ; Wenqiang CHEN
Chinese Journal of Rehabilitation Theory and Practice 2009;15(7):652-653
Objective To investigate the relationship between carotid atherosclerosis (CAS) and cognitive impairment, stroke-specific quality of life of patients with cerebral infarction. Methods Patients enrolled were divided into mild-to-moderate CAS group and severe CAS group after carotid ultrasound. They were assessed with Mini Mental State Examination (MMSE) and Stroke Impact Scale (SIS) Version 3.0. Results The scores of attention/calculation, comprehension, and total of MMSE, the mobile ability and hand function dimensions of SIS were all lower in severe CAS group than in mild-to-moderate CAS group (P<0.05). Conclusion Carotid atherosclerosis may further impair the cognitive function and quality of life of cerebral infarction patients.


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