Objective To study the chemical constituents in the leaves of Nelumbo nucifera. MethodsCompounds were repeatedly purified by chromatography and structures were elucidated by physicochemical properties and spectroscopic analyses. Results Nine alkaloids were isolated and identified as 2-hydroxy-1-methoxy-6-methyl-6a,7-dehydroaporphine (1),armepavine (2),dehydroroemerine (3),dehydronuciferine (4),2-hydroxy-1-methoxyaporphine (5),liriodenine (6),pronuciferine (7),roemerine (8),and nuciferine (9). Conclusion compound 1 is a new aporphine alkaloid named nelumnucine.

Objective To investigate the best protocol of liver 31 P MR spectroscopy . Methods The 31 P MRS was performed using 1.5T MR scanner,heart/liver surface coil and prone position with Resp/Trigger gate in the normal volunteer and the cases with liver diseases including hepatic cirrhosis, hepatocellular carcinoma and benige hepatic tumour. The age ranged From 15 to 58 years old. The related data were processed and compared using several kinds of protocol methods.Results By using the edit skills with Filter, Zero-filling and Curve fitting under the type of J-coupling peak, the curves corresponding with the request and precise data had been obtained for 31 P MRS of the liver. By the simple protocol methods, the 31 P MRS, concentration and ratio of the materials in the liver could be obtained in all cases.Conclusion The process of MR spectroscopy is easy to apply. The requested curves and the accurate data will be obtained by using the editing skill of edit.

Objective: To study the adscription of plasma effective constituents of rat after oral administration of alkaloid fraction of Nelumbo nucifera.Methods: Based on the established HPLC analytical method of plasma effective constituents,analysis and comparison were carried out among HPLC profi les of plasma samples obtained after oral administration of alkaloid fraction of Nelumbo nucifera,blank plasma,alkaloid fraction of Nelumbo nucifera.Results: Sixteen compounds were detected under the method,three of them were architypes of compound contained in alkaloid fraction of Nelumbo nucifera,and the others were metabolites.Through the comparison of the UV spectra,seven metabolites remain essential structure because the max wavelength was same to the architypes’,the other six metabolites’ structure changed greatly.Conclusion: Through the study of the adscription of plasma effective constituents of rat after oral administration of alkaloid fraction of Nelumbo nucifera,the structure of metabolites can be conjectured,which may be helpful to study the ADME and mechanism of action.

AIM:Activin A,a member of transforming growth factor superfamily,is the negative regulator factor in liver regeneration. In this study,the effects of extract of Ginkgo Biloba on hepatic fibrosis and the expression of Activin A in rats with cirrhosis were investigated. METHODS:The experiment was performed at in the Central Laboratory of Wuhan First Hospital from September 2005 to December 2006. ①Thirty-six male SD rats of(160?20) g were randomized into 3 groups:control group,model group and treatment group. ② Except the rats in the control group,others were intraperitoneally injected with 500 mL/L CCl4 for 8 weeks to establish models of hepatic fibrosis. Meanwhile,the extract treatment group was infused with the extract of Ginkgo Biloba(Chinese drugs preparation laboratory of Wuhan First Hospital,detected by Hubeu Wushi Medicine Industry Co.,Ltd. No. 02-391) daily for 8 weeks. ③After administration,all anesthetized rats were sacrificed. Blood samples were collected for the determination of liver function biochemical indexes. Liver tissue samples were used for histopathological examinations. The expression of Activin A was determined by immunohistochemistry and RT-PCR. RESULTS:All 36 rats were involved in the final analysis. ①The liver function in extract treatment group was significantly improved compared with that in model group. ②The grade of fibrosis in extract treatment group were remarkably lower than that in model group under light microscope. ③The positive staining of Activin A in treatment group was significantly reduced compared with model group. ④The expression of Activin A mRNA in extract treatment group was significantly reduced compared with model group. CONCLUSION:Extract of Ginkgo Biloba can effectively decrease the expression of Activin A in rats with hepatic fibrosis caused by CCl4,and lessen the degree of hepatic fibrosis.

OBJECTIVETo discuss the clinical effects of external fixator combined with limited internal fixation in the treatment of pediatric distal femur fractures.

METHODSFrom January 2008 to June 2014, 17 children of distal femur fractures were treated by external fixator combined with limited internal fixation. There were 12 males and 5 females, aged from 6 to 13 years old with an average of 10.2 years, ranged in the course of disease from 1 h to 2 d. Preoperative diagnoses were confirmed by X-ray films in all children. There were 11 patients with supracondylar fracture , and 6 patients with intercondylar comminuted fracture. According to AO/ASIF classification, 9 fractures were type A1, 5 cases were type A2,and 3 cases were type C1. The intraoperative and postoperative complications, postoperative radiological examination, lower limbs length and motion of knee joints were observed. Knee joint function was assessed by KSS score.

RESULTSAll the patients were followed up from 6 to 38 months with an average of 24.4 months. No nerve or blood vessel injury was found. One case complicated with the external fixation loosening, 2 cases with the infection of pin hole and 3 cases with the leg length discrepancy. Knee joint mobility and length measurement (compared with the contralateral), the average limited inflexion was 10 degrees (0 degrees to 20 degrees), the average limited straight was 4 degrees (0 degrees to 10), the average varus or valgus angle was 3 degrees (0 degrees to 5 degrees). KSS of the injured side was (96.4 +/- 5.0) points at final follow-up, 16 cases got excellent results and 1 good. All fractures obtained healing and no epiphyseal closed early was found.

CONCLUSIONExternal fixator combined with limited internal fixation has advantages of simple operation, reliable fixation, early functional exercise in treating pediatric distal femurs fractures.

Adolescent ; Child ; External Fixators ; Female ; Femoral Fractures ; surgery ; Femur ; injuries ; surgery ; Fracture Fixation, Internal ; Humans ; Internal Fixators ; Male

Objective: To explore the effects of Cordyceps extract in regulating the imbalance of Th1/Th2 ratio and inhibiting the inflammatory reaction, and to find the theoretical basis of Cordyceps extract for treating asthma in remission stage. Methods: A total of 20 peripheral venous blood samples (3 mL) were collected from 20 asthmatic children during remission stage, and peripheral blood mononuclear cells (PBMCs) were separated by Ficoll method. PBMCs were separated into three groups (blank group, low-dose group and high-dose group). The PBMCs were incubated in vitro for 48 hours in the absence (blank group) or presence (low-dose group and high-dose group) of Cordyceps extract at different concentrations (10, 20 mug/mL). The expressions of interferon-gamma (IFN-gamma), interleukin-4 (IL-4), IL-10, T-box expressed in T cells (T-bet), GATA-binding protein-3 (GATA-3) and forkhead/winged-helix transcription factor-3 (Foxp3) mRNAs in PBMCs were measured by real-time fluorescent quantitative polymerase chain reaction, and the contents of IFN-gamma, IL-4 and IL-10 in supernatants were measured by enzyme-linked immunosorbent assay. Results: The expressions of IFN-gamma mRNA showed no significant differences among the three groups. The expressions of the IL-4 mRNA in the high-dose group and the low-dose group were lower than that in the blank group (P=0.014, P=0.011). The expression of IL-10 mRNA in the high-dose group was higher than that in the blank group (P=0.034). And the differences of the IFN-gamma mRNA/IL-4 mRNA ratio presented no statistic significance among the three groups. The level of IL-4 content in the high-dose group was lower than that in the blank group (P=0.018), but the level of IL-10 content, and ratio of IFN-gamma/IL-4 in the high-dose group were higher than those in the blank group (P=0.011, P=0.045). The differences of the IFN-gamma presented no statistic significance among the three groups. The T-bet/GATA-3 ratio and Foxp3 mRNA expression in the high-dose group were higher than those in the blank group (P=0.001, P=0.015). There was significant difference in expression of GATA-3 mRNA between the high-dose group and the blank group (P=0.028), and between the low-dose group and the blank group (P=0.019). The expression differences of T-bet mRNA were insignificant between any two groups. Conclusion: Cordyceps extract can inhibit the proliferation and differentiation of Th2 cells and reduce the expression of related cytokines by down-regulating the expression of GATA-3 mRNA and up-regulating the expression of Foxp3 mRNA in PBMCs. Meanwhile, it can alleviate the chronic allergic inflammation by increasing the content of IL-10.

Objective To investigate the diagnostic value of conventional MRI combined with DWI in the differential diagnosis of orbital lymphoproliferative disorders (LPD). Methods As a retrospective study, 42 patients were enrolled, including 23 orbital lymphoma and 19 benign orbital LPD confirmed pathologically or clinically. These patients underwent conventional MR, DWI and contrast?enhanced MRI examination of orbit. Qualitative MR imaging features [location, margin, patient ratio of involved quadrants (≥2), signal intensity,“flow void sign”, sinusitis] and quantitative features [ADC and contrast enhanced ratio of the lesion to temporal muscle (CER)] were evaluated. Chi?square test and t test were used for the analysis of qualitative and quantitative features between lymphoma and benign orbital LPD groups, respectively. Multivariate Logistic regression analysis was employed to identify the significant variable for predicting malignant LPD. ROC analysis was used to evaluate the ability of the diagnostic models established based on identified variables. Results “Flow void sign”(9 lymphomas and 17 benign LPDs) and sinusitis (5 lymphomas and 13 benign LPDs) were statistically significant (P<0.05). The MR imaging features [location, margin, patient ratio of involved quadrants (≥2), and signal intensity] had no significant difference between the two groups (P>0.05). Both ADC [(0.79±0.09)×10?3 mm2/s of lymphoma, (1.29±0.35)×10?3 mm2/s of benign LPD] and CER (1.43±0.17 of lymphoma, 1.79±0.31 of benign LPD) between the two groups had significant difference (t=-6.630, -8.257;P<0.01). Multivariate logistic regression analysis showed that ADC value, CER and“flow void sign”were significant variables for predicting malignant orbital LPD (P<0.05). ROC curve showed that ADC value was the most significant single variable in differentiating orbital LPD [threshold value, 0.74 × 10- 3 mm2/s; areas under ROC curve (AUC), 0.97; sensitivity, 87.0%; specificity, 94.7%]. Combination of ADC and CER could further improve the specificity in differentiating benign from malignant orbital LPD (AUC, 0.90;sensitivity, 78.3%;specificity, 100.0%, accuracy 88.1%). Conclusions ADC, CER and“flow void sign”on T2WI were significant variables for predicting malignant orbital LPD. ADC value was the most significant single variable. Combination of ADC and CER could further improve the specificity in differentiating benign from malignant orbital LPD.

Objective To explore the effect and mechanism of fragile site WWOX gene on regulating proliferation of gallbladder cancer cells in vitro. Methods The pcDNA3.0 - WWOX recombinant plasmid which was previous successfully built was transfected to GBC-SD cells and empty carrier by liposome medium. Liposome and GBC-SD were served as the negative control and the blank control,respectively. After 48 hours transfection, inverted microscope was used to observe the changes of gallbladder cancer cells' morphology,MTT and BrdU were used to detect the proliferation level of gallbladder cancer cells,and flow cytometry instrument was used to detect the change of the cell proliferation cycle. Results The results of inverted microscope shown: the number of GBC-SD cells in pcDNA3.0-WWOX group decreased significantly,the suspension cells and cell debris increased,while cells in the vector control,NC and Mock groups were in normal proliferation state. MTT test showed the proliferation levels of GBC-SD cells in pcDNA3.0-WWOX group was lower than those in the control group in 24 h,48 h,72 h,96 h and 120 h,and the differences were statistically significant(P < 0.05). The cell proliferation activity in the pcDNA3.0-WWOX group was obviously inhibited over time. BrdU detection results showed the cell proliferation rate of pcDNA3.0 - WWOX group was(0.44±0.03),while that in the three control groups was(0.78±0.02), (0.81±0.01)and(0.85±0.01),respectively. It showed that cell proliferation activity in pcDNA3.0-WWOX group was lower than the control groups,and the difference was statistically significant(P < 0.05). Cell cycle detection showed the cells increased in G0/G1 phase and decreased in G2/M and S phases of pcDNA3.0-WWOX group. The cell apoptosis rate was significantly higher and the proliferation index was significantly lower than those of the control groups(P < 0.05). However,there were no significant differences among the three control groups(P > 0.05). Conclusion The overexpression of WWOX gene in vitro could effectively inhibit the proliferation activity of gallbladder cancer cells. WWOX might participate in the development of the malignant biological behavior of gallbladder cancer cells. It is expected to become a new potential target for the gene therapy to gallbladder cancer.

Objective To investigate whether miR?485?3p plays a role in regulation of radiosensitivity of gastric cancer cells by targeting TLR1. Methods Quantitative real?time PCR and Western blot were used to determine the expression of miR?485?3p and TLR1, respectively. The interaction between miR?485?3p and TLR1 was verified by target prediction software ( DIANA, TargetScan, and miRanda) and dual luciferase reporter assay. Gastric cancer MGC803 cells transfected with miR?485?3p mimic or TLR1 siRNA were exposed to irradiation. Apoptosis assay, colony formation assay, and MTT assay were used to evaluate the changes in radiosensitivity of gastric cancer cells. Dual luciferase reporter assay was used to determine the effects of miR?485?3p overexpression and TLR1 silencing on the activity of NF?κB. Western blot was used to study the effects of miR?485?3p overexpression and TLR1 silencing on NF?κB target genes. Results In gastric cancer cells exposed to radiation, the expression of miR?485?3p was downregulated and the expression of TLR1 was upregulated. TLR1 was predicted to be the target of miR?485?3p by target prediction software. Dual luciferase reporter assay further confirmed TLR1 as the direct target of miR?485?3p. miR?485?3p negatively regulated the expression of TLR1. The overexpression of miR?485?3p, as well as TLR1 silencing, increased the apoptosis rate of cells, reduced colony formation and cell proliferation, and enhanced the radiosensitivity of the cells. Both miR?485?3p overexpression and TLR1 silencing reduced the activity of NF?κB and downregulated the expression of multiple NF?κB target genes. Conclusions miR?485?3p enhances the radiosensitivity of gastric cancer cells probably by targeting TLR1 and regulating the NF?κB signaling pathway.

Objective To compare the clinical and ultrasound features of Klebsiella pneumoniae liver abscess(KPLA) in dia‐betes mellitus(DM ) patients with different levels of HbA1c .Methods Totally 146 patients with diabetes mellitus of Klebsiella pneumoniae liver abscesses(KPLA‐DM )were divided into three groups on the basis of their levels of HbA1c:complete glycemic controlled (HbA1c≤7% );good glycemic controlled (7% < HbA1c≤9% );bad glycemic controlled (HbA1c>9% ) .The patients′characteristics ,clinical features ,ultrasound features and complications among the three groups were compared .Results Compared with patients in complete glycemic controlled group and good glycemic controlled group ,patients in bad glycemic controlled group had lower onset age and longer length of hospital stay(P<0 .05) ,are more susceptible to underlying diseases such as hyperlipoi‐demia and chronic renal failure(P<0 .05) ,had more life‐threatening clinical crisis ,and had higher infection rate(P<0 .05) .KPLA‐DM could be characterized by patchy ,mass like enhancement echoultrasound image .Patients in bad glycemic controlled group al‐so had higher risk of biliary pneumatosis ,hepatic venous thrombosis and gas‐forming pyogenic liver abscess than other two groups (P<0 .05) .Conclusion The ultrasound image of KPLA has characteristics .The occurrence of hepatic venous thrombosis and gas‐forming observed by ultrasound is associated with bad glucose control in KPLA‐DM patients ,which suggests that there may be oth‐er complications such as metastatic infection .