Objective To explore the appropriate operative methods for the treatment of CBD(common bile duct) stone in endoscopic era.Methods We retrospectively analyzed the diagnosed and treated data of 309 patients with suspected CBD stones with ERCP,LC+ERCP and LECBD(laparoscopic exploration of the commonbile duct) from January 2004 to July 2008 in our hospital.Results A total of 216 patients receivedERCP,among them 97(44.9%) cases had CBD stone,and there was the trend that the number of patients who received ERCP reduced yearly.Among the 93 patients who received LECBD,71 cases were successful and 22 cases were converted to open operation.Of the 71 cases,transcystic duct CBD exploration was done in 11 cases,direct CBD exploration in 60 cases,and 6 cases had primary closure of CBD.The number of cases that received LECBD grew steadily with time.There was no difference in successful operative rate,intraoperative bleeding and residual calculi rate between ERCP+LC and LECBD group.The operative time,postoperative complications and length of hospital stay in LECBD.group were significantly lower than those in LC+ERCP group.Conclusions LECBD is better than LC+ERCP in the treatment of CBD stones,but in the endoscopic era,the selection of an individualized treatment approach is the best operative method for the management of CBD stone.

Objective To investigate the effect of different irradiation doses of ultraviolet C ray(UVC)on the expression of basic fibroblast growth factor(bFGF)in granulation tissue of gunshot wound in rabbit limbs.Methods After giving 30mJ/cm2 or 60mJ/cm2 UVC to the gunshot wound of soft tissue of limbs,the expression of bFGF was observed both at the mRNA level and the protein level by the methods of in situ hybridization and immunohistochemistry,respectively.Results On the 7th day after UVC irradiation,the expression of bFGF in UVC-treated groups was significantly higher than that in control group(P

Objective To investigate the prevalence of vitamin D deficiency in medical intensive care unit (ICU) and its relationship with severity of disease and prognosis.Methods A prospective study was performed to evaluate vitamin D status in 216 patients admitted to the medical intensive care unit.The incidence of hypovitaminosis D was observed.Acute Physiology and Chronic Health Evaluation Ⅱ (APACHE Ⅱ) score,days kept in ICU and on ventilator,main laboratory findings,and mortality rate were compared among patients with different serum 25-hydroxyvitamin D [25 (OH) D] levels.Potential risk factors for mortality were analyzed by multivariate logistic regression analysis.Results One hundred and fifty-three patients (70.8％) developed hypovitaminosis D.25 (OH) D deficiency was identified in 95 (44.0％),25 (OH) D insufficiency in 58 (26.8％),and 25 (OH) D sufficiency in 63 (29.2％) patients.APACHE Ⅱ score,positive blood culture,the incidence of multiple organ dysfunction syndrome(MODS),and mortality rate were higher in deficiency group compared with the other two groups [APACHE Ⅱ score:deficiency group 25 (20,28) score,insufficiency group 22 (17,26) score,sufficiency group 19 (18,20) score,P＜0.01 ; positive blood culture:deficiency group 18.9％,insufficiency group 13.8％,sufficiency group 3.2％,P=0.015 ; MODS:deficiency group 48.4％,insufficiency group 43.1％,sufficiency group 25.4％,P=0.025; mortality rate:deficiency group 40％,insufficiency group 24.1％,sufficiency group 15.9％,P =0.003].25 (OH)D levels were negatively correlated with APACHE Ⅱ score and mortality rate (r =-0.325,P＜0.01 ; r=-0.276,P＜0.01,respectively).Analysis by multiple logistic regression demonstrated that 25 (OH) D deficiency (OR =3.005,95 ％ CI 1.321-5.875,P =0.008) was independent risk factor for mortality.Conclusions This study demonstrates that vitamin D deficiency is highly prevalent among patients admitted into ICU.Vitamin D deficiency is associated with disease severity,and seems to be an independent risk factor for mortality.

To investigate the relationship between the expression of early growth response gene 1 (EGR-1) and p38MAPK pathway in the paclitaxel resistance of ovarian carcinoma cells, the effect of p38MAPK inhibitor SB203580 on cell apoptosis was examined by using Hoechst 33258 staining. The intracellular Rh123 (Rhodamine 123) accumulation was detected by the flow cytometry (FCM). The 50% inhibition concentration (IC50) of paclitaxel for A2780/Taxol cells was determined by MTT method. Electrophoretic motility shift assay (EMSA) was employed to examine the EGR-1DNA binding activity. MDR1 and EGR-1 mRNA were assessed by RT-PCR. The expressed of p-gp, phosphorylated p53 and p38 were detected by Western blotting. SB203580 could remarkably promote the apoptosis of A2780/Taxol cells, and the cell apoptosis was in a time-dependent manner. Cellular Rh123 accumulation was increased, and the IC50 of paclitaxel for A2780/Taxol cells was decreased significantly. A2780/Taxol cell line after SB203580 treatment was shown to have a significantly higher level of EGR-1 DNA binding activity. SB203580 down-regulated the activity of p38MAPK pathway, but up-regulated EGR-1 expression. SB203580 significantly increased the level of cellular phosphorylated p53 protein, but decreased the p-gp protein level and MDR1 mRNA level in A2780/Taxol cells. There existed a close relationship between p38MAPK pathway and the paclitaxel resistance of ovarian carcinoma cells. The expression of EGR-1 mediated by p38MAPK pathway plays a critical role in paclitaxel resistance of ovarian carcinoma cells.

Aim To investigate whether the pain modi-fication by group I metabotropic glutamate receptors (mGluRs)required the involvement of Src homology-2 domain-containing phosphatase-2 (SHP-2 ).Methods Co-immunoprecipitation was performed to examine the possible interaction between SHP-2 and group I mGluRs in spinal cord dorsal horn of mice.By measur-ing the paw withdrawal thresholds,the effects of SHP-2 inhibitor NSC-87877 or its catalytically inactive SHP-2 (C459S ) mutant on allodynia induced by group I mGluRs agonist DHPG (50 nmol)were observed.Re-sults Anti-mGluR5 antibody was able to co-immuno-precipitate SHP-2 from spinal dorsal horn of mice, while no SHP-2 was precipitated by anti-mGluR1 anti-body.Inactivation of SHP-2 by NSC-87877 (6 nmol) or SHP-2 (C459S ) effectively attenuated allodynia caused by DHPG.Conclusion SHP-2 can physically interact with mGluR5.The activation of SHP-2 may be necessary for group I mGluRs to process the nocicep-tive information.

Objective To construct recombinant lentivirus with the gene STAT3 of the Mus musculus,measure the expres-sion of STAT3,and conduct lentivirus packing and identification.Methods The mRNA of mouse myoblast was extracted and transformed into STAT3 cDNA by the special primer.then,STAT3 cDNA was amplified and reclaimed and inseted into pLVX-IRES-ZsGreen1 vector.Cleavage map and sequencing analysis were used for identification of the recombinant lentivirus vector (pLVX-IRES-ZsGreen1-STAT3).293 T cells were transfected with main vector pLVX-IRES-ZsGreen1-STAT3.and 48 h later, Western blott detected the expression of STAT3 protein.Lentiviral vectors were packaged and the titer was determined.Results The lentiviral vector plasmid pLVX-IRES-ZsGreen1-STAT3 was identified correctly by cleavage map and Co-transfection of 293 T cells with 48 h,the expression of STAT3 was significantly enhanced by western blot.And DNA sequencing analysis confirmed that STAT3 gene sequencing was exactly the same with that reported by genbank.Conclusion Lentiviral vector carrying STAT3 was successfnlly constructed and could express STAT3 with high efficiency,and can be used in further study.

OBJECTIVE: Using the method of lactate dehydrogenase (LDH) assay, to observe the activities of rat cerebral microvascular endothelial cells (CMECs) intervened by Tongluo Jiunao Injection (TLJNI), a traditional Chinese compound drug removing toxin to dredge brain collaterals, and then further study the effects of different kinds of conditioned mediums (CMECs-CM) of cerebral microvascular endothelial cells on ischemia and ischemia/reperfusion cerebral cortex cells, and to probe into the drug pharmacological mechanisms of CMECs in modulating the neurons. METHODS: Three kinds of CMECs (normal, ischemic and ischemic/reperfusional) were all treated by TLJNI previously, and then the three pairs of CMECs-CM without serum were collected respectively for LDH assay. Rat cerebral cortex neurons were also primarily cultured and then divided into similar three groups (normal, ischemic and ischemic/reperfusional). The neuron responses caused by CMECs-CM at different concentrations were observed by using LDH transudation rate assay. RESULTS: The LDH release values of ischemic and ischemic/reperfusional CMECs with TLJNI treatment were obviously reduced (P<0.01) compared with the same kinds of CMECs untreated. For ischemic neurons, both conditioned medium of ischemic CMECs (Is-CM) and conditioned medium of ischemic CMECs with drug treatment (IsT-CM) in high concentration of 100% increased the LDH transudation rate (P<0.01), while in low concentration of 10%, IsT-CM reduced the transudation rate (P<0.05). For ischemia/reperfusion neurons, all kinds of CMECs-CM reduced the transudation rate respectively (P<0.05 or P<0.01). As far as each group concentration was concerned, 10% or 50% showed relatively stronger effects, and both conditioned medium of normal CMECs (N-CM) group and conditioned medium of ischemic/reperfusional CMECs (Rp-CM) group had statistical significance (P<0.05 or P<0.01). For normal neurons, all kinds of CMECs-CM increased the transudation rate respectively (P<0.05 or P<0.01). As far as each group concentration was concerned, only conditioned medium of normal CMECs (N-CM) had statistical significance (P<0.05 or P<0.01). CONCLUSION: The study shows that TLJNI is capable of preventing the damage of CMECs from both ischemia and ischemia/reperfusion states. Chinese drug can restrain the brain ischemia and ischemia/reperfusion damage by the media that CMECs modulate the neurons, demonstrating the pharmacological mechanisms of TLJNI. This work also indicates that there exist some active substances against ischemia/reperfusion injury secreted from CMECs-CM with TLJNI treatment.

Objective To study the feasibility of TNF-α promoting migration of rat mesenchymal stem cells (MSCs) to local damaged tissues. Methods The MSCs was exposed to TNF-α at different concentrations and the expression rate of surface adheslon molecules and specific markers as well as their adhesion to endothelial cells were detected.Based on the above steps,the MSCs stimulated with the optimal concentration of TNF-α were obtained and were injected intravenously to the rats whose hindlimbs experienced ischemia damage.The rats were executed for achieving the muscle samples in the ischemic area,which were made into frozen section to count the number of MSCs. Results ( 1 ) Twenty-four hours after the TNF-o stimulation,the expression of adhesion molecule (VCAM-1) of MSCs increased in a concentration-dependent manner,while the expression of adhesion molecules (ICAM-1,L-Selectin and VLA4) of MSCs showed no significant changes.Besides,the expression rate of specific markers of MSCs was also obscure.(2) Exposed to 10 ng/ml TNF-o,MSCs presented an obviously increased ability in adhesion to the endothelial cells.(3) MSCs stimulated with 10 ng/ml TNF-α showed a larger number in the ischemia-damaged tissue of rat hindlimbs than that in the control group. Conclusion TNF-α at concentration of 10 ng/ml is effective within a short term in increasing VCAM-1 expression in rat MSCs and promoting the adhesion of MSCs to endothelial cells without affecting their character.

Objective To investigate the changes of fecal short-chain fatty acids (SCFA) and bile acid levels in patients with colon cancer.Methods Totally 189 patients with colon cancer (CC group),201 patients with adenomatous polyp (AP group),and 512 healthy patients (control group) who were confirmed by endoscopy were included in this study.The fecal SCFA and bile acid levels were measured by enzyme linked immunosorbent assay.Results The total bile acids,primary bile acids,and secondary bile acids were not significantly different among these three groups (P ＞ 0.05).The chenodeoxycholate level in the CC group [0.338 (0.101,0.416) mg/g] was significandy higher than that in AP group [0.241 (0.108,0.375) mg/g] and control group [0.248 (0.110,0.371) mg/g] (P=0.025,P=0.023),but was not significantly different between the AP groupand the control group (P ＞ 0.05).The deoxycholic acid level in CC group [0.375 (0.136,0.503) mg/g] and AP group [0.369 (0.113,0.494) mg/g] were significandy higher than that in control group [0.277 (0.115,0.412) mg/g] (P=0.026,P=0.024),and the difference between CC group and AP group was not statistically significant (P ＞ 0.05).The level of lithocholic acid in CC group [0.386 (0.147,0.507) mg/g] was significantly higher than those in the AP group [0.103 (0.012,0.238) mg/g] and control group [0.239 (0.081,0.405) rng/g] (P=0.011,P=0.027); also,its level in AP group was significantly lower than that in the control group (P =0.022).The levels of total short-chain fatty acids,acetic acid,propionic acid,and isovaleric acid were not significantly different among the control group,AP group,and CC group (P＞0.05).The levels of butyrate [0.105 (0.059,0.198) mg/g,0.090 (0.050,0.183) mg/g],isobutyl acid [0.036 (0.024,0.046) mg/g,0.025 (0.020,0.034) mg/g] in CC group and AP group were significantly higher than in the control group [0.081 (0.051,0.107) mg/g,0.021 (0.016,0.029) mg/g] (butyrate:P=0.026,P=0021; isobutyl acid:P=0.025,P=0.019),and the difference between CC group and AP group was statistically significant (butyrate:P =0.031; isobutyl acid:P =0.024).Conclusions Fetal chenodeoxycholic acid,lithocholic acid,butyric acid,and isobutyric acid may play a role in the developmem of colon cancer,while deoxycholic acid may also be implicated in both colon cancer and colon adenomas.No association is found between other SCFA and bile acids and colorectal cancer/adenoma.

Objective To observe and compare the atomic force microscopy (AFM) images of Yersinia pestis EV76 and the changes in the morphology of the bacteria treated with normal serum and F1 antibody from rabbit,and to explore the immunoassay method to detect Yersinia pestis by AFM. Methods The Yersinia pestis were treated with normal serum and F1 antibody from rabbit and control buffer. All the prepared samples were observed and analyzed by AFM. The changes in the cell surface structures were probed and characterized through sectional analysis,especially the changes of Ra and Rq value. Results The normal morphology of Yersinia pestis was oval in shape with a relatively smooth surface, the size dimension of which was about 1.1-1.3 μm in length with a section profile of 0.8-1.0 μm in width and 0.04-0.06 μm in step height. The step height of the bacteria treated with the normal serum and F1 antibody was obviously enlarged. The shape of the bacteria treated with F1 antibody changed irregularly. Furthermore, the surface of the bacteria was more roughened. Conclusion The morphological characters of Yersinia pestis has been acquired through its AFM images. The morphology of Yersinia pestis treated with F1 antibody has changed greatly, and the index of roughness can be regarded as the distinguished index to detect Yersinia pestis by AFM.