1.Overexpression of Sox9 gene by the lentiviral vector in rabbit bone marrow mesenchymal stem cells for promoting the repair of cartilage defect.
Zhen WANG ; Da-chuan LIANG ; Jie-yu BAI ; Ning KANG ; Jun-yu FENG ; Zi-quan YANG
China Journal of Orthopaedics and Traumatology 2015;28(5):433-440
OBJECTIVETo study the overexpression of Sox9 gene on rabbit bone marrow mesenchymal stem cells for repairing articular cartilage injury in vivo.
METHODSRabbit bone marrow mesenchymal stem cells (BMSCs) were transduced with lentivirus vector containing Sox9 gene and then cartilage specific molecule was detected by RT-PCR in vitro. Total 48 knee joints of 24 mature New Zealand white rabbits were randomly divided into 3 groups according to different defect treatment. After animals anesthesia,a full-thickness cylindrical cartilage defect of 4 mm diameter and 3 mm deep was created in the patellar groove using a stainlesssteel punch. Meanwhile, the transfected cells were implanted to repair the rabbit model with full-thickness cartilage defects. Cartilage defects tissue was observed with light microscope, electron microscope, HE and immunohistochemistry staining to assess the repair of defects by the complex at 6 weeks or 12 weeks after the implantation.
RESULTSAt 3 days after the transfection, Sox9 gene expression was highest and Sox9 gene expression decreased with the increase of time. At 3 days after the transfection, the expression of collagen type II began and reached the peak at 14 days. It showed that the bone marrow mesenchymal stem cells went into chondrogenic differentiation after transfected by Sox9 gene. Histological observation showed that at 6 weeks after the operation, the defects in the experimental group was filled with hyaline like cartilage tissue, 12 weeks after operation,the defects of cartilage and subchondral bone had satisfactory healing. Both at 6 and 12 weeks postoperatively, the defects were filled with fibrous tissues in control groups. Meanwhile, immunohistochemical staining of sections with type II collagen antibodies showed the proteins in the regenerated tissue stained positive for type II collagen and stronger than the control groups. The histological scoring system indicated that the cartilage repair of experiment groups were better than the two control groups with statistical significances.
CONCLUSIONOverexpression of Sox9 gene on rabbit bone marrow mesenchymal stem cells (BMSCs) promote the repair of cartilage defect.
Animals ; Bone Marrow Cells ; metabolism ; Bone Marrow Transplantation ; Cartilage, Articular ; injuries ; metabolism ; Cell- and Tissue-Based Therapy ; Female ; Genetic Vectors ; genetics ; metabolism ; Humans ; Lentivirus ; genetics ; metabolism ; Male ; Mesenchymal Stem Cell Transplantation ; Mesenchymal Stromal Cells ; metabolism ; Osteoarthritis ; genetics ; metabolism ; therapy ; Rabbits ; SOX9 Transcription Factor ; genetics ; metabolism ; Tissue Engineering
2.Male pseudohermaphroditism due to 17β-hydroxysteroid dehydrogenase 3 deficiency
Jun YANG ; Guang NING ; Lihao SUN ; Jie HONG ; Jialun CHEN ; Manyin XU ; Weiqing WANG ; Xiaoying LI
Chinese Journal of Endocrinology and Metabolism 2008;24(3):272-274
Objective To investigate the clinical and genetic characteristics in a patient with 17β-hydroxy-steroid dehydrogenase (17β-HSD) 3 deficiency, regarding its pathophysiology and pathogenesis. Methods Clinical features and laboratory data were analyzed in a pedigree of 17β-HSD3 deficiency. Blood samples from the patient and his parents were collected. HSD17B3 gene was screened for mutations by PCR and subclone sequencing. Results The patient presented with pubertal virilization and gynecomastia. The physical examination showed female external genitalia and testes in inguinal canals. The chromosome karyotype was 46, XY. Serum FSH, LH, dehydroepiandrosterone sulfate, androstenedione and 17-OH-progesterone levels were raised, whereas plasma testosterone was lowered. Sequencing analysis revealed 4 nucleotide deletion (172-175del) of HSD17B3 gene. Conclusion Virilization and gynecomastia in puberty suggest the probability of 17β-HSD deficiency. It may be verified clinically by hCG-stimulating test and confirmed by gene diagnosis.
3.Mechanism of varied responsiveness to ACTH in patients with polycystic ovary syndrome
Cheng XU ; Jun YANG ; Huijie ZHANG ; Jie HONG ; Guang NING ; Xiaoying LI
Chinese Journal of Endocrinology and Metabolism 2010;26(5):359-363
Objective To explore the potential mechanism of adrenal androgen excess in patients with polycystic ovary syndrome (PCOS), ACTH stimulation test was conducted and the polymorphisms in the promoter region of CYP21 A2 gene were screened to verify the variations related to the responsiveness to ACTH stimulation. Methods 30 healthy women and 101 PCOS patients, matched for age, were recruited from Ruijin hospital. Blood biochemical examinations were taken and sex hormone profiles obtained at baseline. 17 hydroxyprogesterone( 17OHP)was measured at 0 and 60 min in an ACTH stimulation test. The -710 bp -1 bp of the promoter region of CYP21A2 was sequenced in 87 PCOS patients and 30 control subjects. Results According to the post-stimulation 17 OHP levels obtained from 30 healthy women,PCOS patients were allocated into one group with high responsiveness to ACTH ( HR-PCOS, n = 21) and the other with normal responsiveness to ACTH ( NR-PCOS, n = 80). Compared with NR-PCOS subjects, HR-PCOS patients had higher testosterone( P<0.05), basal and post-stimulation 17OHP (both P<0.01)and dehydroepiandrosterone sulfate levels (P<0.05 or P<0.01) .whereas serum cortisol and androstenedione levels were not significantly different before and after ACTH stimulation test. The genotypes of locus -535 were well correlated with post-stimulation 17OHP levels (r = 0. 20,P = 0. 03) in PCOS patients and the control subjects. The genotype T/T or allele T was significantly more frequent in subjects with a higher fertile of post-stimulation 17OHP (P<0.05 or P<0. 01). The odds ratio(OR)for higher responsiveness to ACTH in women with allele T at -535 was 3. 69 (95% CI 1. 69-8. 06,P = 0. 000 7). Conclusions The PCOS patients with higher responsiveness to ACTH are characterized by severe hyperandrogenemia and adrenal androgenexcess,suggesting that adrenal androgen excess in some PCOS patients may be due to higher responsiveness to ACTH. The polymorphism of -535C>T in the promoter region of CYP21 A2 may play a role in regulating 21 hydroxylase gene expression and further influencing 17OHP responsiveness to ACTH.
4.Relationship between survivin expression and paclitaxel drug-resistance and research of reversed drug-resistance by matrine in non-small cell lung cancer
Su-Xia LUO ; Xiao-Bing CHEN ; Ning LI ; Jun-Jie ZENG ;
Cancer Research and Clinic 2006;0(12):-
Objective To study the correlation between drug-resistance of paclitaxel and the expres- sion levels of anti-apoptotic protein survivin and the role of matrine reversal resistance of PTX in non-small cell lung cancer.Methods The expressions of survivin in 120 samples of human lung cancer with paclitaxel chemotherapy were detected by immunohistochemical staining.Results The positive expression rate of sur- vivin was 57.5%.In the positive expression of survivin group,chemotherapy combined matrine could improve response rate and survival time(P0.05).Conclusion The survivin expression was related to the response rate of paclitaxel in lung cancer.It might be a valuable new marker to predict the drug-resistance of paclitaxel.In this prospective research,survivin protein,which promoted the apoptosis caused by paclitaxel,may be the target of martine.It could partly reverse the drug re- sistance to paclitaxel.
5.Application of team based learning in teaching of dental endodontics
Wenming HUANG ; Jie QIN ; Hui XUE ; Ning GAN ; Jun LUO ; Deqin YANG
Chinese Journal of Medical Education Research 2012;(12):1229-1232
Objective To apply team based learning (TBL) in the teaching of dental endodontics for undergraduates in order to expand and deepen the TBL teaching in stomatology education.Methods TBL was used among 2008 grade oral undergraduates in Chongqing Medical University.Average test scores of 2008 grade undergraduates before and intra class were calculated and questionnaire was designed.At the same time,final examination scores between 2007 grade and 2008 grade under-graduates were compared.Results There was no significant correlation in average test scores before and intra class (r =0.027,P > 0.05).Final examination scores were higher in 2008 grade than in 2007 grade.Based on the questionnaire survey,learning interests,sense of teamwork and classroom knowledge grasp of 2008 grade undergraduates were obviously elevated.Conclusions TBL teaching significantly improve students' learning effect and it can be promoted in stomotology education.
6.Observation on clinical effect of tuina plus Western medication for functional dyspepsia due to liver qi stagnation and spleen deficiency
Jiong-Jie NING ; Jun YU ; Wu LI ; Jiang-Shan LI ; Guo-Qi HUANG
Journal of Acupuncture and Tuina Science 2018;16(2):109-114
Objective:To observe the clinical effects of tuina plus Western medication for functional dyspepsia (FD) due to liver qi stagnation and spleen deficiency.Methods:A total of 72 patients in conformity with the inclusion criteria of FD were randomly divided into an observation group and a control group based upon the random number table,36 cases in each group.The control group was treated with mosapride citrate dispersible tablets,and the observation group was treated with the same tablets plus tuina.Before the treatment and 4 weeks after the treatment,the clinical symptoms,quality of life (QOL) and depression severity were observed by the scale,and were followed up two months later after the treatment for assessment of the clinical effects.Results:After the treatment and at the follow-up,the symptom scores of FD and the sores of Hamilton depression rating scale (HAMD) in both groups decreased,and the scores in Chinese version of quality of life questionnaire for functional digestive disorders (Chin-FDDQL) increased,with statistically significant differences in comparison with the same group before the treatment (all P<0.05).In comparison between the two groups at the same time point after the treatment,the scores of FD symptoms,HAMD and Chin-FDDQL were improved better in the observation group than those in the control group,with statistically significant differences (all P<0.05).The total effective rates at the follow-up were 91.7% in the observation group and 75.0% in the control group,without statistical difference between the two groups (P>0.05).The rate of clinical cure and remarkable effect was 66.7% in the observation group,higher than 41.7% in the control group,it is higher in the observation group than that in the control group,with a statistically significant difference between the two groups (P<0.05).Conclusion:Tuina plus Western medication is precise in the therapeutic effects for FD due to liver qi stagnation and spleen deficiency and can effectively relieve clinical symptoms,elevate the QOL and alleviate depression severity of the patients.Moreover,it's better than the treatment by Western medication alone in the long-term therapeutic effects.
8.Relationship between chronic prostatitis and prostatic calculus.
Hong-jie CHEN ; Ning-gang YANG ; Ju-jie ZHANG ; Jun WANG ; Xiang-jun ZHANG ; Jun ZHANG ; Xin-ning YU ; Dian-ting ZHANG
National Journal of Andrology 2011;17(1):43-46
OBJECTIVETo explore the relationship between chronic prostatitis (CP) and prostatic calculus (PC).
METHODSWe used transperineal ultrasonography (TPUS) to detect PC in 500 normal volunteers and 491 CP patients, and divided them into a CP and a CP + PC group according to the ultrasonographic results. Then we analyzed the NIH-CPSI scores, duration of symptoms and white blood cell count in the expressed prostate secretion (ESP).
RESULTSPC was found in 19.8% of the normal controls, 5% (5/100), 12% (12/100), 19% (19/100), 27% (27/100) and 36% (36/100) in the 20-30 yr, 31-40 yr, 41-50 yr, 51-60 yr and > 60 yr groups, respectively. In comparison, PC was detected in 42.2% of the CP patients, 15.8% (12/76), 30.1% (69/215), 55.7% (59/109), 66.2% (43/65) and 82.8% (24/29) in the above five age groups, respectively, with statistically significant differences between the control and CP groups (P < 0.01). The CP and CP + PC groups showed significant differences in the duration of symptoms and white blood cell count in ESP (P < 0.01) but not in CPSI scores (P < 0.05).
CONCLUSIONThe incidence of PC is higher in CP patients than in healthy men, and it is associated with inflammation, aging and symptom duration, but not with CPSI scores.
Adult ; Aged ; Aged, 80 and over ; Bodily Secretions ; Calculi ; pathology ; Case-Control Studies ; Chronic Disease ; Humans ; Inflammation ; Leukocyte Count ; Male ; Middle Aged ; Prostate ; secretion ; Prostatic Diseases ; pathology ; Prostatitis ; pathology ; Young Adult
9.Effect of thymosin alpha1 on immunological function and metabolism in peritoneal sepsis rats.
Jun JIANG ; Ning LI ; Jie-Shou LI
Chinese Journal of Surgery 2004;42(22):1377-1380
OBJECTIVETo investigate the effect of thymosin alpha1 on immunological function and protein metabolism in peritoneal sepsis rats.
METHODSWe observed the effect of thymosin alpha1 on T cell subclassification, TNFalpha, IL-6, IL-10, albumin, C-reactive protein (CRP) and mortality in cecal ligation and puncture (CLP) induced septic rats.
RESULTSConcentration of TNFalpha and IL-6 in CLP induced septic rats increased significantly, and concentration of IL-10 decreased significantly compared to control group. Thymosin alpha1 significantly decreased TNFalpha, IL-6, and significantly raised concentration of plasma IL-10, percent of CD(3), CD(4), and CD(4)/CD(8) in septic group. Thymosin alpha1 reduced degressive degree of albumin. Concentration of CRP increased in both septic groups, but was less prominently in thymosin alpha1 treated group. Thymosin alpha1 reduced cumulate 7-day mortality.
CONCLUSIONThymosin alpha1 can improve immunological function, inflammation condition and protein metabolism.
Adjuvants, Immunologic ; pharmacology ; Animals ; C-Reactive Protein ; metabolism ; Disease Models, Animal ; Interleukin-10 ; blood ; Interleukin-6 ; blood ; Peritonitis ; drug therapy ; immunology ; metabolism ; Rats ; Rats, Sprague-Dawley ; Sepsis ; drug therapy ; immunology ; metabolism ; T-Lymphocyte Subsets ; immunology ; Thymosin ; analogs & derivatives ; pharmacology ; Tumor Necrosis Factor-alpha ; metabolism