Two patients with neonatal diabetes tested as V59A and V59M mutations were chosen for the study. Clinical data were analyzed retrospectively. The results showed that the patient with V59A mutation was characteristic of spasm and hyperglycemia at the age of three month, and treated with insulin for a long time as unresponsive to the glibenclamide at the beginning. Myasthenia and delay of development were observed during the follow-up. At the age of two years, glibenclamide was tried for the second time with a high dose and fairly-controlled glucose level. The patient with V59M mutation was diagnosed with diarrhea, hyperglycemia, and ketosis at the age of two month, and was responsive to glibenclamide at a relatively low dose with well-controlled glucose level. These results suggest that KCNJ11 V59M mutation would show some milder clinical manifestations and better glibenclamide efficacy as compared with V59A mutation.

Objective To study the protective effect and its mechanism of anti-tumor necrosis factor-?antibody (TNF-? Ab) on lung injury after cardiopulmanary bypass(CPB).Methods Twenty-eight healthy rabbits were selected and randomly divided into four groups:group Ⅰ only received open chest operation;groups Ⅱ-Ⅳ underwent CPB.In the group Ⅳ,rabbit TNF-? Ab (2 400 pg/kg) was dropped into the intracheal tube before operation and just after releasing the aortic clamp.Saline was given to the group Ⅲ in- stead.Blood neutrophils count,TNF-?,MDA from the right and left atrium in the four groups were determined perioperatively.Water volume,TNF-? mRNA,TNF-? protein,apoptosis and pathomorphological changes were measured in the lung tissues.Results TNF- ? Ab can restrain leukocyte accumulation,reduce releasing of TNF-? and MDA in the lung.It can also reduce the occurrence of apop- tosis and attenuate pathomorphological changes in the lung tissue.However,it cannot reduce the secretion of TNF-? at the transcrip- tion level and protein level.Conclusion Intratracheal TNF-? Ab administration has markedly protective effect on lung injury after CPB.

Objective To investigate the efficacy and toxicity of neoadjuvant regional arterial chemotherapy in the treatment of advanced gastric cancer. Methods The clinical data of 158 patients with advanced gastric cancer and with the same clinical stages who were admitted to Renji Hospital of Shanghai Jiaotong University from February 2002 to May 2005 were retrospectively analysed. Preoperative regional arterial chemotherapy was applied to 76 patients (test group) and the remaining 82 patients only received surgical treatment (control group). The chemotherapy regimen was epirubicin (50 mg/m2) + cisplatin (60 mg/m2) + 5-fluorouracil (1000 mg/m2).This regimen was modified to oxaliplatin (130 mg/m2) + 5-fluorouracil (1000 mg/m2) since 2003, and surgery was performed 6-11 days after the chemotherapy. All patients received postoperative intravenous chemotherapy.The clinical effects, radical resection rate, operative complications and long-term survival of the two treatment methods were evaluated. All data were analysed using the chi-square test and Kaplan-Meier analysis. Results The radical resection rate was significantly higher at 86% (65/76) in the test group compared with 71% (58/82)in the control group ( x2 = 5.01, P ＜ 0. 05 ). The toxicity of the chemotherapy in the test group was mild. The postoperative complication rate was 20% (15/76) in the test group and 16% (13/82) in the control group, with no significant difference between the two groups (x2 = 0.41, P＞0.05). The median survival time was 41 months in the test group and 23 months in the control group. The 5-year overall survival rate was higher in the test group (44.6%) than that in the control group (29.1%) (x2 =3.95, P＜0. 05). Conclusions Neoadjuvant regional arterial chemotherapy is well tolerated by patients with advanced gastric cancer. It is also effective for increasing the radical resection rate and improving the long-term survival.

Objective To investigate the preopera ti ve diagnosis and the surgical treatment of primary retroperitoneal tumor(PRT). Methods The clinical dat a of 71 patients with PRT were retrospectively analyzed including clinical manif estation, radiologic studies, pathologic examination and surgical procedures. Results There were 32 cases of benign tumor, 38 cases of malignant tumor and 1 case of borderline tumor. Of the 32 patients with benign tumor, 31 underwent complete surgical resection. Of the 38 patients with malignant tumor, 29 underwent complete resection. One patie nt with borderline tumor underwent total resection. The 5-year survival rate of benign PRT was 89.29%, the 5-year survival rate of malignant PRT was 20.80%. Four cases of recurrent benign PRT underwent complete resection. Ten cases of r ecurrent malignant PRT underwent complete resection and 3 underwent partial rese ction. ConclusionsRadiolog ical study is crucial for the diagnosis. Complete resection is the key for the t reatment of PRT.

OBJECTIVETo investigate the expression changes and regulation of pituitary adenylate cyclase-activating polypeptide (PACAP) mRNA in corpus luteum during pregnancy.

METHODSPregnant rats' ovaries were collected at different time points. The techniques of RT-PCR and in situ hybridization were used to observe expression changes of PACAP mRNA in rat ovaries during pregnancy. To further explore the regulation mechanism of PACAP mRNA expression in corpus luteum, luteal cells were cultured in vitro. Immature (25 - 28 days old) female Sprague-Dawley rats were injected subcutaneously with 50IU pregnant mare serum gonadotrophin (PMSG), and 25IU human chorionic gonadotrophin (hCG) 48 h later, to induce follicular development and luteum formation. On day 6 after hCG administration (the day of hCG administration was the first day), the rats were killed by guillotine and the ovarian luteal cells were collected. After incubation for 24 h, luteal cells were administration with various factors for 24 h. And then expression changes of PACAP mRNA in luteal cells after administration with different factors were detected by RT-PCR, and radioimmunoassay was used to analyze progesterone levels.

RESULTSWith the development of pregnancy, the expression of PACAP mRNA increased gradually, reached the peak at pregnancy 19 d, and then decreased. Compared with control group, platelet activating factor (PAF), forskolin and PMA could obviously stimulate PACAP mRNA expression in luteal cells which were cultured with corresponding factors for 24 h. At the same time, progesterone levels in culture media were also elevated.

CONCLUSIONPACAP, acting as a local ovary regulator, was closely related to the maintenance of medium-term and late pregnancy. PAF could directly stimulate PACAP mRNA expression in luteal cells, and protein kinase C (PKC) and protein kinase A (PKA) signal pathways could both participate in this process.

Animals ; Cells, Cultured ; Corpus Luteum ; metabolism ; Female ; Pituitary Adenylate Cyclase-Activating Polypeptide ; genetics ; metabolism ; Platelet Activating Factor ; metabolism ; Pregnancy ; RNA, Messenger ; genetics ; Rats ; Rats, Sprague-Dawley

Objective To develop a HPLC method for determination of pueratin.Methods The separation was carried out on a Waters Symmetry C18 column(4.6 mm×250 mm, 5 μm), the mobile phase was composed of acetonitrile and 1% formic acid(11∶89), the detection wavelength was set at 250 nm, the flow rate was 1.0 ml/min, the column temperature was 30 ℃ and the injection volume was 10 μl.Results The linearity was obtained over 2~40 μg/ml (r=0.999 8) for pueratin.The RSD of precision were less than 2%.The average recovery was between 98% and 103%.Conclusion This HPLC method was simple, accuracy and suitable for the quality control of Jiangzhi Hugan capsule.

Background Retinitis pigmentosa (RP)is a common hereditary blinding eye disease in ophthalmology.Current researches documented that RP may have the common pathophysiologic basis to Alzheimer disease and chronic neurodegenerative disease.Understanding this mechanism will offer a new therapeutic target for RP.Objective The purpose of the present study was to investigate the roles of cyclin-dependent kinase 5 (Cdk5)/P25 activation in the apoptosis of retinal neural cells of RCS rats.Methods Eighteen SPF RCS rats and 18 RCS-rdy+ rats were randomized into 17-,25-and 35-day groups respectively and 6 rats for each.The rats were sacrificed at corresponding time points and retinal hemogenete was prepared.Expressions of CdkS,P35,P25 and tau phosphorylation in the retinas were detected by Western blot,and the kinase activity of Cdk5/P25 was analyzed by quantitative colorimetric assay.Results The expressing level of P35 protein(A340) in the retinas of 17-day-old RCS rats was near that of 17-day-old RCS-rdy+ rats(t =0.52,P＞0.05).In 25-and 35-day-old RCS rats,the expressing levels of P35 protein were 2.20±0.48 and 1.23±0.14,which were higher than those of RCS-rdy+ rats(1.43±0.13 and 0.93±0.10),showing significant differences between them(t =3.78,4.28,P＜0.05).The expression of P25 was undetectable at postnatal 17 days in RCS rats and RCS-rdy+ rats,but it showed significantly higher in RCS rats(0.300±0.003 and 0.230±0.004) than that in RCS-rdy+ rats(0.040±0.004 and 0.070±0.004) at postnatal 25 days and 35 days(t=121.81,77.51,P＜0.01).No significant difference was found in the expression of Cdk5 in RCS rats and RCS-rdy+ rats at different ages (t =-0.60,0.19,1.62,P＞ 0.05).The kinase activity of Cdk5/P25 did not show significantly different between RCS and RCS-rdy+ rats at postnatal 17 days(t =0.19,P＞0.05),but significantly higher kinase activity of Cdk5/P25 was seen in RCS rats (0.0058 ±0.0005 and 0.0056±0.0004) than that in RCS-rdy+ rats(0.0038±0.0003 and 0.0032 ±0.0007) at postnatal 25 days and 35 days (t =8.07,5.97,P＜ 0.01).No expression of tau phosphorylation was detected in RCS rats at postnatal 17 days,but significantly higher tau phosphorylation level was seen in RCS rats at postnatal 25 days and 35 days(1.80±0.22 and 1.23±0.17),which were significant different in comparison with RCS-rdy+ rats at postnatal 25 days and 35 days(1.60 ±0.20 and 1.04 ±0.12)(t=4.71,3.17,P＜0.05).Conclusions The Cdk5/P25 kinase activity shows a consistent trend with theexpressions of P25 and tau phosphorylation in the RCS rats,indicating that the upregulation of P25 induces the enhance of enzyme activity of Cdk5,which phosphorylate its substrates to result in more apoptosis of retinal neural cells.

Objective To predict and identify liner B-cell epitopes in the hemagglutinin ( HA) of human-infected avian-origin H7N9 influenza virus and analyze the specificity of H7 subtype.Methods Three serum samples collected at different times from the same patient who was confirmed to be infected with H7N9 influenza virus were provided by Shaoxing People’s Hospital, and one serum sample from healthy person was collected as the control.The extracellular region of HA protein was predicted by TMHMM Sever v.2.0.The potential B-cell epitopes were predicted by DNAStar Lasergene’ s Protean, BcePred and ABCpred tools, and the immunogenicity of the predicted B cell antigen epitopes was assessed by indirect enzyme-linked immunosordent assay ( ELISA ) .H7 subtype specificity was analyzed by comparing HA protein amino acid sequence with H7N9 and H1-H16 subtype influenza virus from Genbank using Clustal X 2.1 software, and Cn3D 4.3.1 software was used to detect the distribution and 3D structure of predicted epitopes on the HA protein of H7N9.Results The potential B-cell epitopes may be located in 172-183, 363-380, 452-472 and 491-506 of extracellular N-terminus of HA protein.ELISA showed that four predicted eptiopes specifically reacted with positive serums from patient.Multi-sequence alignment demonstrated that peptide 172-183 and 363-380 had higher H7 subtype specificity compared with amino acid sequences of other subtypes.Moreover, the predicted linear B-cell epitopes all located on the surface of HA protein according to the 3D structure analysis.Conclusion Four potential B-cell epitopes were identified, in which peptide 172-183 and 363-380 have higher H7 subtype specificity, and may be used in the design of epitope-based vaccines and diagnostics tests.

A role of gut microbiota in colorectal cancer (CRC) growth was first suggested in germ-free rats almost 50 years ago, and the existence of disease-associated bacteria (termed pathobionts) had becoming increasingly evident from experimental data of fecal transplantation, and microbial gavage or monoassociation. Altered bacterial compositions in fecal and mucosal specimens were observed in CRC patients compared to healthy subjects. Microbial fluctuations were found at various cancer stages; an increase of bacterial diversity was noted in the adenoma specimens, while a reduction of bacterial richness was documented in CRC samples. The bacterial species enriched in the human cancerous tissues included Escherichia coli, Fusobacterium nucleatum, and enterotoxigenic Bacteroides fragilis. The causal relationship of gut bacteria in tumorigenesis was established by introducing particular bacterial strains in in situ mouse CRC models. Detailed experimental protocols of bacterial gavage and the advantages and caveats of different experimental models are summarized in this review. The microbial genotoxins, enterotoxins, and virulence factors implicated in the mechanisms of bacteria-driven tumorigenesis are described. In conclusion, intestinal microbiota is involved in colon tumorigenesis. Bacteria-targeting intervention would be the next challenge for CRC.
Adenoma ; Animals ; Bacteria ; Bacteroides fragilis ; Carcinogenesis* ; Colon ; Colorectal Neoplasms ; Enterotoxins ; Escherichia coli ; Fecal Microbiota Transplantation ; Fusobacterium nucleatum ; Gastrointestinal Microbiome ; Healthy Volunteers ; Humans ; Mice ; Microbiota* ; Models, Theoretical* ; Mutagens ; Rats ; Virulence ; Virulence Factors

The present study was undertaken to investigate the effects of ceramide on progesterone production and apoptosis in rat luteal cells in vitro. Luteal cells were prepared from the ovaries of pregnant mare serum gonadotropin (PMSG) and human chorionic gonadotropin (hCG) primed female Wistar rats and incubated with cell permeable C2-ceramide. The effects of ceramide on progesterone production and apoptosis in luteal cells were assessed by radio immunoassay (RIA) and flow cytometry analysis. In addition, changes in nitric oxide synthase (NOS) activity and nitric oxide (NO) production by luteal cells treated with C2-ceramide were also evaluated. Ceramide was found to reduce hCG-stimulated progesterone production in a dose-dependent manner, whereas it had little effect on basal progesterone content. Spontaneous apoptosis in luteal cells was observed after a 12-hour incubation in vitro and 5 μmol/L of ceramide significantly increased the apoptotic rate (P<0.05). Enhanced apoptotic peak was seen in the histogram by flow cytometry. Moreover, 50 μmol/L of C2-ceramide significantly increased NOS activity (P<0.01) and NO production (P<0.001). It is suggested that ceramide may serve as an important signaling molecule mediating certain ovarian processes, such as luteal regression.