Objective To summarize the diagnosis and therapeutic experience of insulinoma in order to improve the surgical success rate and prognosis. Methods The clinical data of 138 patients with insulinoma from 1966 to 2007 were retrospectively analyzed. Results In this group of patients, hypoglycemia of different levels and Whipple triad were detected. 64 patients expressed different psychic symptom, 12 patients' psychic symptom were still present after blood glucose normalized after operation. Fasting serum insulin values in 88 patients were measured, and the insulin release index was higher than 0. 3. Before operation, tumor was detected in 8 of 75 patients by B-ultrasound scan, and in 17 of 68 patients by CT, and in 5 of 10 patients by MRI. The intra-operative B-ultrasound (IOUS) examination was applied in 44 cases, and 43 cases were successfully detected. The operations included enucleation of insulinoma (n=88) , resection of the body and tail of pancreass (n = 44) , pancreaticoduodenectomy (n=2) , and biopsy (n=1). The blood glucose symptoms normalized postoperatively in 132 patients. The blood glucose rebound in 110 patients, but blood glucose normalized within 2 weeks. After operation, 20 patients developed pancreatic fistula, 32 patients developed acute pancreatitis. Conclusions Insulinoma could be qualitatively diagnosed according to Whipple triad and the insulin release index. Operations with IOUS were simple and effective methods to localize the tumors. The only way to cure insulinoma was operation, and IOUS guided operation could avoid main pancreatic duct and vessel injury, decrease post-operative complications.

Objective To investigate the action of chondrogenesis differentiation of bone marrow stromal cells (BMSCs) transfected with adeno-hTGF-β1. Methods In the experiment group, replication-deficient a denoviruses carrying human hTGF-β1 complementary DNA (adeno-hTGF-β1 was constructed and applied to transfect to the first generation BMSCs. As a control, each BMSCs was transduced with 200 pfu of adeno-LacZ gene. One day after transfer, BMSCs were trypsinized, counted, and 5×105 cells aliuots were spun down at 500 rpm per minute in 15 ml polypropylene conical tubes and then cultured in a defined medium in an incubator at 37℃ for 21 days. The aggregates were harvested at time points to 21 days and assessed by gross observation, histological analyses and immunohistochemical localization of type Ⅱ collagen. Results When harvested at 21 days, each pellet shrinked to spheroid tissue with apearly opalescence in gross morphology and found to be relatively firm. H.E staining showed elongate dlining cells appeared as perichon drium-like cells at the surface. Some nests of cartilage were observed at the substrate of the tissue. Mature chon drocytes were embeded in the lacuna in the experiment group. In addition, Safranin'O staining confirmed the presence of sulfated proteoglycans in the ECM of chondrogenesis region. Immunohistochemical staining revealed the presence of type Ⅱ collagen in chondrogenesis region. By contrast, HE staining showed no evidence of cartilage formation in the control group. They were fibrous tissue with no architectural feature. Safranin'O staining and Immunohistochemical staining showed no evidence of sulfated proteoglycans or typeⅡ collagen expression. Conclusion BMSCs transfected with adeno-hTGF-β1 could induce its chondro-genesis when aggregate cultured in a defined medium in vitro, laying a foundation for the application of hTGFβ1 gene-transfected BMSCs in cartilage tissue engineering.

Objective To summarize the experience in the diagnosis and management of insulinoma. Methods From January 1966 to December 2007, the clinical data of 131 patients with insulinoma were retrospectively analyzed. Results All the 131 cases had Whipple triad syndrome and 64 eases suffered from psychoneurosis symptoms. The fasting blood glucose or insultus blood glucose concentration of all the cases was lower than 2.8 mmol/L. The ratios of serum insulin to glucose were all higher than 0.3. Before operation, tumor was detected in 8 of 75 patients by B-us scan, and in 17 of 68 by CT, and in 5 of 10 by MRI. The intraoperative ultrasonography(IOUS) was applied in 44 eases, and tumor was found in 43 cases. Surgery included enucleation of insulinoma (88 cases), resection of the corpus and eauda of the pancreas (40 cases), duodenopancreatectomy (2 cases), and biopsy (1 case). The low blood glucose symptoms disappeared postoperatively in 130 cases. Pancreatic fistulae occurred in 20 cases, acute pancreatitis occurred in 32 cases. Conclusions Insulinoma can be diagnosed based on symptoms of Whipple triad and the ratio of serum insulin to glucose. Exploration and IOUS are the simple and effective methods to localize insulinoma.

To develop a new small molecular probe for discovering an antitumor lead compound from the replacement of carboxylic group of two molecular antibacterial fluoroquinolones with a heterocyclic ring, a series of the C3/C3 bis-fluoroquinolones tethered with an 1, 3, 4-oxadiazole ring were synthesized as their respective HCl salts, and their structures were characterized by elemental analysis and spectral data. The in vitro antitumor activity against L1210, CHO and HL60 cell lines was also evaluated via the respective IC50 values by methylthiazole trazolium (MTT) assay.

Objective To report a ileal ureteric replacement surgery for bilateral long segment ureter injury.Methods Data from a patient suffered bilateral long segment uretercic stenosis was reviewed.A 32-year-old female,who underwent radical surgery of cervical cancer and postoperative radiotherapy 1 year ago,complained intermittent low back pain for 11 months and was found renal function abnormality for 3 months.Ultrasound showed bilateral hydronephrosis,and the CT urography showed bilateral middle-lower ureteral stenosis.Preoperative diagnosis was bilateral hydronephrosis with bilateral ureteral obstruction.The patient underwent bilateral ileal ureteric replacement under general anesthesia.Results The surgical procedure was successful and the postoperative recovery was favorable.The operation time was 180 min,and blood loss was 100 ml.The abdominal cavity drainage tube was removed 8 days and the urinary catheter was removed 9 days postoperatively.During 8 months' follow up,the patient showed resolved flank pain with stable serum creatinine.No complication was reported.The CTU 2 months postoperatively showed the hydronephrosis was relieved with normal functioning kidneys.Conclusion Bilateral ileal ureteric replacement surgery might be an effective procedure and viable option for bilateral long segment ureteric injuries.

AIM:To observe the expression of nuclear factor-kappa B ( NF-κB) , N-methyl-D-aspartic acid re-ceptor 2B (NR2B) and inducible nitric oxide synthase (iNOS) in the spinal cord in a rat model of chronic constriction in-jury (CCI) of the sciatic nerve.METHODS:Fifty-six adult male Sprague-Dawley rats weighing 180~220 g were random-ly divided into sham group (n=8) and CCI group (n=48).The mechanical withdrawal threshold (MWT) and paw with-drawal latency (PWL) of the hind paws were measured 1 d before CCI and 1 d, 4 d, 7 d, 14 d and 21 d after surgery.The L4~L6 segment of the spinal cord was taken for determining the expression of NF-κB, NR2B and iNOS by RT-PCR and Western blotting.RESULTS:At 1 d, 4 d, 7 d, 14 d and 21 d after surgery, the MWT and PWL in CCI group were obviously lower than those in sham group .The expression of NF-κB, NR2B and iNOS at mRNA and protein levels in-creased significantly.Positive correlations were found between the mRNA expression of NF-κB and iNOS (r=0.842, P<0.05), and between the mRNA expression of NR2B and iNOS (r=0.833, P<0.05).CONCLUSION:The generation and maintenance of hyperalgesia in sciatic nerve injury rats may attribute to the activation of NF -κB and NR2B and concom-itant increase in iNOS .

Objective To investigate the effect of disodium cantharidinateon in vitro lymphocyte immune response in lung cancer patients.Methods Twenty non-small cell lung cancer patients diagnosed by pathological diagnosis were selected.The effects of different concentrations of disodium cantharidinate and vitamin B6 on lymphocyte proliferation and CD4 + CD25 + T cells,CD25 + FOXP3 + T cells,CD4 + and CD8 + T cells were detected by methyl thiazolyl tetrazolium assay and flow cytometry.Results When the concentrations of disodium cantharidinate and vitamin B6 was less than 10 μg/ml,with the growth of concentration,it stimulated the proliferation of peripheral blood lymphocytes.When the concentration was more than 10 μg/ml,with the growth of concentration,the absorbance value decreased,and the stimulating effect weakened.When the concentration of disodium cantharidinate and vitamin B6 injection was less than 10 μg/ml,with the growth of concentration,CD4 +/CD8 + ratio increased significantly,and CD4 + CD25 +/CD4 + and CD25 + FOXP3 + T cells were significantly lower,which showed statistically significant differences (t =2.171,P =0.032 ; t =2.103,P =0.041 ; t =3.662,P =0.002 ; t =3.201,P =0.003 ; t =3.233,P =0.003).But when the concentration was more than 10 μg/ml,The differences of CD4 +/CD8 + ratio,CD4 + CD25 +/CD4 + and CD25 + FOXP3 + T cells in 10,15,20 μg/ml groups were not statistically significant,which showed that disodium cantharidinate and vitamin B6 injection had a positive effect on enhancing the immune function of lymphocytes in lung cancer in a concentration dependent manner,but high concentration was unhelpful.Conclusion Disodium cantharidinate and vitamin B6 injection can promote lung cancer in vitro lymphocyte proliferation as well as increase its immunity in a certain range.

Objective To study the expression and the clinical significance of hypoxia-induced factor-1α (HIF-1α) in gallbladder cancer tissues,and the role and mechanism of HIF-1α in metformin-suppressed metastasis in gallbladder carcinoma GBC-SD cells.Methods 24 specimens of gallbladder cancer tissues and 5 specimens of chronic cholecystitis were collected from the First Affiliated Hospital of Zhengzhou University between June 2016 and February 2017.Immunohistochemistry and qPCR were used to detect the expression of HIF-1α in gallbladder cancer tissues,in adjacent non-cancer tissues and in chronic cholecystitis,and the clinical significance was analyzed.The model of metastasis was induced by hypoxia;the wound healing assay and the Transwell assay were used to detect the ability of cell metastasis;the expressions of HIF-1α and VEGF in gallbladder carcinoma GBC-SD cells were detected by western blotting assay and immunofiuorescence.Results The expression of HIF-1α in gallbladder cancer tissues was higher than the adjacent non-cancer tissues and in chronic cholecystitis.The expression of HIF-1α was correlated with lymph node metastasis and TNM staging in gallbladder cancer tissues (P ＜ 0.05).The wound healing rate after 48 h in the negative control group and in the treatment with hypoxia group (1％ O2) in GBC-SD cells were (46.5 ± 4.8) ％ and (67.3 ± 4.0) ％,respectively.The Transwell data showed that the numbers of metastasis after 24 h in the negative control group and in the treatment with hypoxia group GBC-SD cells were (147.4 ± 11.7) and (234.4 ± 17.7),respectively.When compared with the negative control group,treatment with hypoxia significantly increased the ability of metastasis and up-regulated the expression of HIF-1α and VEGF in GBC-SD cells (P ＜ 0.05).The wound healing rate after 48 h in the negative control group,the metformin group,the hypoxia group and the metformin and hypoxia group in GBC-SD cells were (40.6 ± 7.1) ％,(16.4 ± 9.4) ％,(69.5 ± 4.0) ％ and (22.4 ± 7.4) ％,respectively.The Transwell data showed that the numbers of metastasis after 24 h in the negative control group,the metformin group,the hypoxia group and the metformin and hypoxia group in GBC-SD cells were (148.4 ± 6.9),(90.0 ± 8.4),(185.8 ± 10.2) and (113.4± 8.6),respectively.When comparcd with the hypoxia group,treatment with metformin and hypoxia significantly decreased the ability of metastasis and down-regulated the expression of HIF-1α and VEGF in GBC-SD cells (P ＜ 0.05).The wound healing rate after 48 h in the negative control group,the 2MeoE2 group,the hypoxia group,the 2MeoE2 and hypoxia group in GBC-SD cells were (43.4 ±4.4)％,(25.9 ±9.0)％,(63.3 ±2.2)％,(46.2 ±4.5)％,respectively.The Transwell data showed that the numbers of metastasis after 24 h in the negative control group,the 2MeoE2 group,the hypoxia group,the 2MeoE2 and hypoxia group in GBC-SD cells were (144.2 ± 12.6),(80.2 ±7.7),(203.8 ±7.0),(124.0 ± 5.2),respectively.When compared with the hypoxia group,treatment with HIF-1α inhibitor 2MeoE2 and hypoxia significantly decreased the ability of metastasis and down-regulated the expression of HIF-1α and VEGF in GBC-SD cells (P ＜ 0.05).Conclusions The expression of HIF-1 α was correlated with lymph node metastasis and TNM staging in gallbladder cancer tissues.Treatment with hypoxia significantly increased the expression of HIF-1α and VEGF and promoted metastasis of GBC-SD cells,while treatment with metformin decreased the ability of metastasis induced by hypoxia via inhibiting the HIF-1o/VEGF pathway in GBC-SD cells.

Objective:To predict Th/B cell epitopes in HA of influenza virus(H1N1)and analyze antigenicity of the candidate epitopes in order to develop epitope-bacterin by the way of bioinformatics.Methods:The HA amino acid sequences of infiuenza virus(H1N1),which the viral infection was prevalent recently,were downloaded from Genbank.The Th/B cell epitopes were predicted and analyzed by bioinformatics methods.Then,specificity and conservation of the candidate epitopes were estimated.Finally,antigenicity of the candidate epitopes was identified by influenza virus(H1N1)positiVe serum samples of mice.Results:Three Th/B cell epitopes containing HA_(73-87),HA_(125-139),HA_(188-205) were acquired Two of the candidate epitopes were in a relatively conserved domain of HA1,and a deal of 2006-2009 influenza virus(H1N1)isolates contained the sequences.Moreover,the candidate epitopes were showedin a distinct antibody combining reactivity with the influenza virus (H1N1)positive serum of mice,which inferred the predicted epitopes to be functional ones.Conclusion:The selected epitopes are able to be functional HA Th/B cell epitopes of influenza virus(H1N1).Our study also establish the foundations for the further research of influenza virus infectlon and immunity mechanism,the recognition of influenza virus(H1N1)functional epitope and the development of epitope vaccines.

Objective:To predict Th/B cell epitopes in HA of influenza virus(H1N1) and analyze antigenicity of the candidate epitopes in order to develop epitope-bacterin by the way of bioinformatics.Methods:The HA amino acid sequences of influenza virus (H1N1),which the viral infection was prevalent recently,were downloaded from Genbank.The Th/B cell epitopes were predicted and analyzed by bioinformatics methods.Then,specificity and conservation of the candidate epitopes were estimated.Finally,antigenicity of the candidate epitopes was identified by influenza virus (H1N1) positive serum samples of mice.Results:Three Th/B cell epitopes containing HA73-87,HA125-139,HA188-205 were acquired.Two of the candidate epitopes were in a relatively conserved domain of HA1,and a deal of 2006-2009 influenza virus (H1N1) isolates contained the sequences.Moreover,the candidate epitopes were showedin a distinct antibody combining reactivity with the influenza virus (H1N1) positive serum of mice,which inferred the predicted epitopes to be functional ones.Conclusion:The selected epitopes are able to be functional HA Th/B cell epitopes of influenza virus (H1N1).Our study also establish the foundations for the further research of influenza virus infection and immunity mechanism,the recognition of influenza virus (H1N1) functional epitope and the development of epitope vaccines.