[Abstract ] Objective Bone marrow mesenchymal stem cells(BMSCs) can be induced to the differentiation into vascular smooth muscle cells in many induction conditions.We sought to explore the possibility of the differentiation of mesenchymal stem cells into vascular smooth muscle cells by continuous cell culture in vitro. Methods Rat BMSCs were isolated from the bilateral tibial and femoral bones by the method of whole bone marrow adherence, followed by ex vivo expansion.BMSCs were identified by flow cytometry and three-lineage differentiation.After continuous five days'cell culture of BMSCs, the specific surface antigens of VSMCs were detec-ted by immunofluorescence, western blot and real-time PCR. Results BMSCs expressed CD29、90, in contrast, they did not express CD45、34、49d.After induction of osteogenesis, adipogenesis and chondrogenesis, alizarin red、oil red and alcian blue staining pro-duced a strong reaction in cells.The expressions ofα-SMA、Calponin1、SM-MHC and SM22 in the cells of experimental group were no-tably increased, which indicated that BMSCs were differentiating towards VSMCs. Conclusion In the absence of exogenous stimula-tion, BMSCs can be successfully induced to differentiate into VSMCs by continuous cell culture.

Objective:To investigate the effect of 3 HLA matching measures on preventing hyperacute rejection(HAR). Methods:Among 1 152 cases,550 cases received anti donor lymphocytotoxic crossmatch(LXM),335 cases received LXM and HLA serological test,267 cases of the latter again received panel reaction antibody(PRA)test, the change of the HAR rate with different HLA matching measures were analyzed. Results:If LXM value ≤10% or HLA matched loci ≥3,the rate of HAR was unrelated to LXM value or HLA matched loci. HAR rate decreased significantly in patients with LXM and HLA serological test, and decreased more markedly in those with additional PRA test compared with those only with LXM test. Conclusion:HLA test and PRA are necessary and effective to avoid HAR.

Objective To investigate the clinical implication of Neoral C 2 in evaluating drug hepatic toxicity following renal transplantation in hepatitis patients. Methods 162 cases receving renal transplantation were divided into two groups: positive group with positive hepatitis B and(or) C viral antigens or de finite hepatitis history before renal transplantation ( n =51) and the remaining 111 cases served as normal control group. One week posttransplantation, the blood concentrations of cyclosporins at 0 (C 0) and 2 (C 2) h after administration were determined. Results In the positive group, C 2, not C 0 one week postoperation was higher than normal control group ( P

Objective To understand the distribution and drug resistance of pathogens from patients with anastomotic fistula infection after esophageal cancer surgery, and provide basis for clinical diagnosis and treatment.Methods Patients were admitted to a hospital due to anastomotic fistula after esophageal cancer surgery between January 2012 and December 2015, microbial culture and antimicrobial susceptibility testing results of patients were retrospectively analyzed.Results 1 986 patients underwent radical resection of esophageal cancer within 4 years, 148 of whom developed anastomotic fistula, 104 (70.27％) were with positive microbial culture.A total of 197 pathogenic strains were isolated, 52(26.40％)and 145 (73.60％)strains were isolated from intrathoracic anastomotic fistula and cervical anastomotic fistula respectively;127 (64.47％)strains were gram-negative bacteria, the major were Pseudomonas aeruginosa, Klebsiella pneumoniae, and Acinetobacter baumannii, 62(31.47％) strains were gram-positive bacteria, the major were Staphylococcus aureus, Enterococcus spp., and Streptococcus viridans, 8 strains (4.06％) were fungi.49(47.12％) cases were with mixed pathogenic infection.The resistance rates of gram-negative bacteria to imipenem were 17.86％-47.62％, to polymyxin B was 0, resistance rates of Pseudomonas aeruginosa to other antimicrobial agents were all<50％, Klebsiella pneumoniae to piperacillin and aztreonam were both>70％, Acinetobacter baumannii to most antimicrobial agents were all>50.00％;resistance rates of gram-positive bacteria to clindamycin and tetracycline were both>50.00％, to linezolid, vancomycin, and teicoplanin were all 0, resistance rates of Staphylococcus aureus to penicillin, oxacillin, and ciprofloxacin were all>60％,resistance rate of Enterococcus spp.to quinupristin/dalfopristin was 100.00％.Conclusion Postoperative anastomotic fistula combined with infection can affect the prognosis of patients after esophageal cancer surgery, regular monitoring on distribution and drug resistance of pathogens can provide the basis for initial empirical treatment, and is conducive to the early treatment and rational use of antimicrobial agents.

Objective To explore the characteristics and method that used modified septoplasty in high posi- tion debiation drawing(or)accompanying dislocation of nosal septum.Methods By performing modified septoplasty on 59 patients who had high position deviation and former dislocation of nosal septum,improvement on method was carried out,and aspects of 59 patients in operation such as data,operation method,follow-up analysis were checked. Results The follow-up visiting in the following 5 years showed that the 59 patients' nasal septum were in the cen- tre of the nose.and there was no dislocation again or no perforation.Conclusion The improved method is simple and convenient,so it can be easily used in clinics.

Objective To investigate the risk factors of cervical esophagogastric anastomotic fistula after esophagectomy of esophageal cancer.Methods The retrospective case-control study was conducted.The clinicopathological data of 956 patients who underwent esophagectomy and cervical esophagogastrostomy from January 2012 to December 2016 in the First Affiliated Hospital of Zhengzhou University were collected.Patients underwent Sweet or Mckeown surgery.Observation indicators:(1) intra-and post-operative situations;(2) the risk factors analysis of cervical esophagogastric anastomotic fistula after esophagectomy;(3) follow-up situations.Follow-up using outpatient examination and telephone interview was performed to detect the esophagogastric anastomotic stenosis of patients up to February 2017.Measurement data with normal distribution were represented as the (x)±-s.Univariate analysis and comparison of count data were done using the chi-square test or Fisher exact probability method.Multivariate analysis was done using the Logistic regression model.Results (1) Intra-and post-operative situations:all the 956 patients underwent successful operations,including 107 with Sweet operation and 849 with Mckeown operation.Of 956 patients,336 received thoracotomy and 620 received thoracoscopic surgery.Tumors located in upper,middle and lower esophagus were respectively detected in 143,627 and 186 patients.Operation time,volume of intraoperative blood loss and number of lymph node dissected in 956 patients were (274 ± 67) minutes,(210 ± 167) mL and 18 ± 11,respectively.Of 956 patients,117 had cervical esophagogastric anastomotic fistula,with an incidence of anastomotic fistula of 12.24％ (117/956).Of 117 patients with cervical esophagogastric anastomotic fistula,2 had early stage fistula,110 had middle stage fistula and 5 had later stage fistula;12 were cured by two-tube method (stomach tube and nutrition tube),24 were cured by three-tube method (stomach tube,nutrition tube and chest tube or mediastinal tube),43 were cured by open neck incision dressing,15 were cured by fistula cavity drainage and 17 were cured by esophageal stent implantation.Sixteen patients died in hospital postoperatively,including 6 with cervical esophagogastric anastomotic fistula and 10 without cervical esophagogastric anastomotic fistula.Duration of hospital stay of 956 patients was (16± 11)days,and durations of hospital stay of patients with and without cervical esophagogastric anastomotic fistula were (39± 19) days and (13±6) days.Postoperative pathological examinations:873,9 and 74 patients were respectively diagnosed with squamous cell carcinoma,adenocarcinoma and other types of cancer.TNM staging:stage 0,Ⅰ,Ⅱ,Ⅲ,Ⅳ and unidentified stage were respectively detected in 135,110,325,376,1 and 10 patients.(2) The risk factors analysis of cervical esophagogastric anastomotic fistula after esophagectomy:univariate analysis showed that gender,age,history of diabetes,surgical method,tubular stomach production,operation time,postoperative pulmonary infection and postoperative aspirating sputum through fiberbronchoscope were risk factors affecting cervical esophagogastric anastomotic fistula after esophagectomy,with statistically significant differences (x2 =4.179,6.174,4.427,4.377,6.266,7.057,55.036,51.806,P＜ 0.05).Multivariate analysis showed that tubular stomach production,postoperative pulmonary infection and aspirating sputum through fiberbronchoscope were independent risk factors affecting cervical esophagogastric anastomotic fistula after esophagectomy,with statistically significant differences (OR =1.922,2.907,2.323,95％ confidence interval:l.203-3.070,1.682-5.023,1.235-4.370,P＜0.05).(3) Follow-up situations:908 of 956 patients were followed up for 2-62 months,with a median follow-up time of 28 months.During the follow up,21 of 111 patients with cervical esophagogastric anastomotic fistula were complicated with cervical esophagogastric anastomotic stenosis,59 of 797 patients without cervical esophagogastric anastomotic fistula were complicated with cervical esophagogastric anastomotic stenosis,showing a statistically significant difference in cervical esophagogastric anastomotic stenosis (x2-16.803,P＜0.05).Conclusion Tubular stomach production,postoperative pulmonary infection,postoperative aspirating sputum through fiberbronchoscope are independent risk factors affecting cervical esophagogastric anastomotic fistula after esophagectomy.

OBJECTIVETo explore the mechanism and feasibility of the supinator syndrome evoked embitterment test from anatomy and clinic.

METHODS25 cases of The supinator syndrome were reviewed. 18 of them were male and 7 were female. Drop finger deformation were apparent in 25 cases and The supinator syndrome evoked embitterment test was positive for All patients. Operative neurolysis was done in 8 cases, conservation treatment 17 cases; 92 cadaver upper extremities were dissected for a study the relationship between supinator tunnel and posterior interosseous nerve.

RESULTS22 cases had been followed up for an average of 9 months. 16 cases had a full recovery and 6 cases, a partial recovery. the anatomical study shows that The posterior interosseous nerve was compressed by Forhse arcade and the distal border of the supinator muscle during passive pronation forearm.

CONCLUSIONThe supinator syndrome evoked embitterment test was a new test for the diagnosis of supinator syndrome, it was found to be more sensitive and specific than the others test.

Exercise Test ; methods ; Female ; Humans ; Male ; Nerve Compression Syndromes ; diagnosis ; Radial Nerve ; pathology ; Radial Neuropathy ; diagnosis ; pathology ; therapy ; Sensitivity and Specificity

OBJECTIVETo observe the protective effects of the Huanglian Jiedu decoction aqueous extract and its active fraction, which consists of total alkaloids, total flavonoids and total iridoid, on the thalamus of cerebral ischemia in rats.

METHODThe rat model of middle cerebral artery occlusion (MCAO) was chosen. Male SD rats were randomly divided into sham-operation group, model group, aqueous extract group (800 mg x kg(-1)), total alkaloids group(44 mg x kg(-1)), total flavonoids group (50 mg x kg(-1)) and the total iridoid group (80 mg x kg(-1)). The rats were administered the appropriate drugs intragastrically once a day, for 7 days after surgery. An equivalent volume of saline was given in the sham surgery and model groups. The HE staining was adopted to observe the pathological changes. Determination of Glu and gamma-GABA in thalamus were detected by HPLC with fluorescence detection. The expression of GAD65 was examined with immunohistochemistry and double staining with uorescent-conjugated antibodies against GFAP and Cx43 was chosen in this study.

RESULTThe neurons degenerated in MCAO rats after cerebral ischemia 7 d. The content of Glu, gamma-GABA decreased (P < 0.05), the expression of GAD65 reduced (P < 0.05) and the expression of GFAP and Cx43 increased (P < 0.01) in thalamus of rats compared with sham-operation group. Huanglian Jiedu decoction aqueous extract, total alkaloids, total flavonoids and total iridoid reduced the degeneration of neurons. Total flavonoids could promote the expression of GAD65 (P < 0.05) and decrease the expression of GFAP and Cx43 (P < 0.01) in thalamus compared with model group while it could also increased the content of Glu,gamma-GA BA to normal levels. Compared with model group, Huanglian Jiedu decoction aqueous extract, total alkaloids and total iridoid could raise the expression of Cx43, and Huanglian Jiedu decoction aqueous extract could also increase the expression of GAD65 (P < 0.05). The expression of GFAP in Huanglian Jiedu decoction aqueous extract group, total alkaloids group and total iridoid group were not different compared with model group while the content of gamma-GABA decreased (P < 0.05) compared with sham-operation group.

CONCLUSIONThe degeneration of nerve cells, the reduction of neurotransmitter amino acids content, the aberrant activation of astrocytes and the abnormal expression of GFAP and Cx43 will appear in thalamus of MCAO rats after ischemia. Huanglian Jiedu decoction total flavonoids could relieve the injury of nerve cell through inhibiting the abnormal activation of astrocytes and regulating the expression of GFAP and GAD65.

Animals ; Astrocytes ; drug effects ; Disease Models, Animal ; Drugs, Chinese Herbal ; pharmacology ; Infarction, Middle Cerebral Artery ; drug therapy ; Male ; Protective Agents ; pharmacology ; Rats ; Rats, Sprague-Dawley ; Thalamus ; drug effects

This study examined the effect of Notch-1 signaling on malignant behaviors of breast cancer cells by regulating breast cancer stem cells (BCSCs). BCSCs were enriched by using serum-free medium and knocked out of Notch-1 by using a lentiviral vector. Real-time polymerase chain reaction (RT-PCR) and Western blotting were used to detect the Notch-1 expression levels in breast cancer cell lines and BCSCs, and flow cytometry to detect the proportion of BCSCs in BCSC spheres. The BCSC self-renewal, migration, invasion, and tumorigenicity were examined by the tumor microsphere-forming assay and transwell assay and after xenotransplantation. The results showed that the Notch-1 silencing reduced the number of BCSC spheres, the proportion of BCSCs, and the number of cells penetrating through the transwell membrane. It also decreased the size of tumors that were implanted in the nude mice. These results suggest that Notch-1 signaling is intimately linked to the behaviors of BCSCs. Blocking Notch-1 signaling can inhibit the malignant behaviors of BCSCs, which may provide a promising therapeutical approach for breast cancer.

OBJECTIVETo examine the effect of pcDNA3.1-VEGF165 vector to the angiogoiesis, expression of collagen type I and type III mRNA in soft tissue injury model.

METHODSThirty two Sprague-Daulay rats,weighted (180 +/- 20) g, were made tissue injury in the bilateral of vertebral region. Round wound (diameter 12 mm) was made by perforex on the back, removed the skin and 2 mm muscle, one side was experimental group by random and the other as control. The wound was done with sodium chloride (0.2 ml) in the control group, with the recombinant VEGF165 vector (0.2 ml, 200 mg) in the experimental group. The wound healing and other general state of health was observed after the operation. The specimens were obtained at 3,5, 7,14 and 30 days after injury. Draw the materials from the rats at the same time, all samples were divided into two parts. one ( > 0.1 g) was conserved in refrigerator at - 80 degrees C, which was extracted total RNA by TRIZOL, design the primer of rat's collagen type I and type III, RT-PCR analysis indicated that collagen type I, III. The other was fixed by 10% formalin. Examine wound healing of local tissue and count it' s MVD by HE staining.

RESULTSAll the rabbits were well alive, no death or infection. Wound healing time was shorter than the control one (14.2, 17.4 d). Inflammatory cell infiltrate, cellula intersitialis, fibroblast, collagen and the density of angiogenesis were more in the experimental group than in the control one. The MVD was significant difference between the two groups at 1, 2 weeks are 63.38 +/- 9.20, 52.72 +/- 7.06 and 76.64 +/- 12.27, 66.84 +/- 9.82 (P < 0.05). The expression of collagen type I , III mRNA was found in the third day, the peak was in the second week and then degression. The collagen type I , III mRNA and beta-actin specificitic belt were found and its initial template volume different, the results was trend of RT-PCR obtained.

CONCLUSIONSThe local application of pcDNA3.1-VEGF165 can enhance the expression of collagen type I, III mRNA, enhance angiogenesis and extra cellular matrix, both of which can shorten healing time of tissue injury.

Animals ; Collagen Type I ; metabolism ; Collagen Type III ; metabolism ; Genetic Vectors ; RNA, Messenger ; metabolism ; Rats ; Rats, Sprague-Dawley ; Soft Tissue Injuries ; genetics ; metabolism ; Transfection ; Vascular Endothelial Growth Factor A ; genetics ; Wound Healing