Abstract: A new selaginellin derivative named as selaginellin S (1) was isolated from the whole plants of Selaginella pulvinata (Hook. et Grev.) Maxim. (Selaginellaceae), together with a known one (selaginellin M, 2). Compounds 1 and 2 were separated and purified by silica gel and Sephadex LH-20 column chromatography. Their structures were determined on the basis of extensive spectroscopic analysis including IR, MS, 1D and 2D NMR experiments, as well as ECD calculations. Compound 1 is a key intermidiant in the biosynthesis pathway of selaginellins. Compound 2 is first reported in this plant.

OBJECTIVE To investigate the distribution and resistance of fungi isolated from hemopathy patients,and to provide the laboratorial data in order to prevent and treat infection caused by fungi more effectively.METHODS Strains were isolated from the upper respiratory tract,lower respiratory tract,genitourinary tract,alimentary tract;and perianal region,perineum,and the soft tissue and skin wounds.Antimicrobial susceptibility of clinical isolates was tested by broth microdilution susceptibility test.RESULTS Candida albicans,Aspergillus,tropicalis,C.krusei,and C.glabrata were the main fungi of total 3104 detected isolates.The resistance rates of C.albicans and C.tropicalis to fluconazole,itraconazole and 5-flucytosine were no more than 1.4%.The MIC50-MIC90 range of C.tropicalis for itraconazole was narrow(0.25-0.5 mg/L),but that was wide for fluconazole(0.25-4.0 mg/L).CONCLUSIONS C.albicans,Aspergillus,C.tropicalis,C.krusei,and C.glabrata are possibly the most common pathogens causing endogenous and exogenous fungi infection.The main infection sites of Aspergillus are the upper respiratory tract and lower respiratory tract,which may be associated possibly with ward's air quality.The MIC50to MIC90 range of C.tropicalis is wider for fluconazole than for itraconazole,which is possibly correlated with overexpression of CtMDR1 in C.tropicalis caused by the widely application of fluconazole in therapy.

Objective To explore the characteristics and the clinical application of MR perfusion weighted imaging(PWI)in the normal kidneys and the renal diseases.Methods Thirty-one subjects including 9 cases without urinary diseases,14 cases with renal carcinoma,6 cases with renal cyst and 2 cases witll renal tuberculosis who had been examined with T1WI,T2WI and PWI were analyzed retrospectively.All the data were processed by a workstation to obtain time-signal intensity curves,color perfusion maps and relative perfusion value.The relative renal blood volume(RBV),relative renal blood flow(RBF),mean transition time(MTT)and the time to peak(TTP)in the normal renal cortex and medulla and the renal lesions were calculated.Comparisons between the right and the left normal kidneys,and between the cortex and the medulla of the normal kidneys were performed using t test,and comparisons between the normal and the abnormal kidneys were performed using q test.Results Relative RBV and relative RBF of the cortex were 1.33±0.08 and 1.44±0.09 respectively,and for medulla were 0.58±0.05 and 0.78±0.13 respectively(t=9.2241 and 5.0336,P＜0.01);MTT(1.11±0.08)and TTP (1.04±0.06)of the cortex has no difference with that of the medulla(0.97±0.04 and 0.94±0.03)in the normal kidneys(t=2.2551 and 2.2613.P＞0.05).The values of relative RBF of the renal carcinoma (1.35±0.34)were significantly higher than that of the normal tissues(1.02±0.06)(q=3.0882,P＜0.01).Conclusion PWI is able to demonstrate the hemodynamic change of the normal renal tissues and the renal lesions,and it maybe an ideal method for showing the functional changes of the kidney and for differentiating the renal diseases.

OBJECTIVE:To establish the quality standard of Jingyuan capsule. METHODS:TLC was used for qualitative iden-tification of Epimedium brevicornu,Pseudostellaria heterophylla and Fallopia multiflora in the preparation. HPLC method was used to determine the content of icariin. The determination was performed on Odyssil C18 column with mobile phase consisting of acetoni-trile-water(28:72,V/V)at the flow rate of 1.0 mL/min. The detection wavelength was set at 270 nm,and the column temperature was 30 ℃. The sample size was 10 μL. The contents of polysaccharides was determined by phenol-sulfuric method. RESULTS:TCL spots of E. brevicornu,P. heterophylla and F. multiflora were clear and well separated without interference from negative con-trol. The linear range of icariin ranged 0.027-0.135 mg/mL(r=0.9999);RSDs of precision,stability and repeatability test were all lower than 2.0%. The recoveries were 97.87%-101.94%(RSD=1.47%,n=9). The linear range of glucose ranged 0.056-0.121 mg/mL(r=0.9995),and RSDs of precision,stability and repeatability tests were all lower than 2.0%. The recoveries were 99.37%-100.38%(RSD=0.36%,n=6). CONCLUSIONS:The established standard can be used for the quality control of Jingyuan capsule.

Objective To identify hepatitis C virus(HCV)-specific cytotoxicity T lymphocytes (CTL)epitopes by the combination of T epitopes prediction software and in vitro assays.Methods HCVspecific CTL epitopes were predicted by T epitope prediction software Rankpep and then candidate HCV-specific CTL epitopes were selected.Candidate HCV-specific CTL epitopes were used to stimulate PBMC of HCV-infected patients and healthy volunteers.and then enzyme-linked immunospot(ELISPOT)and intracellular cytokine staining(ICS)were used to measure the frequencies of IFN-γ-producing cells in total PBMC and the percentages of IFN-γ+CD8+T cells in total CD8+T cells,respectively.Results Five candidate CTL epitopes[NS3 450(TVPQDAVSR),NS3 594(GPTLLYRL),Ns4b 78(sMMAFSAAL),NS5a 416(SEENVSVVF)and NS5a 367(TVSSALAEL)]were used to stimulate PBMC of ten HCV-infected patients and two healthy volunteers.PBMC of seven HCV-infected patients secreted IFN-γ while PBMC of healthy volunteers did not produce IFN-γ.The frequencies of peptide-specific IFN-γ-producing cells ranged from 5 to 36 SFC/105 PBMC and the percentages of peptide-specific IFN-γ+CD8+T cells ranged from 0.02%-0.25%.Conclusion Resuhs of ELISPOT assay and ICS assay confirm that these five peptides NS3 450,NS3 594,NS4b 78,NS5a 416 and NS5a 367 are identified as Hovel HCV-specific CTL epitopes.

Objective To study the relationship between viral infection and chronic obstructive pulmonary disease (COPD),and the effect of viral infections on the changes of Th1/Th2 in COPD patients. Methods The se-ra from 81 patients with acute exacerbations of COPD,25 patients with stable COPD and 22 healthy subjects were tested for specific IgM of respiratary syncytial virus(RSV) ,herpes simplex virus type 1 (HSV-1), parainfluenza vi-rus (PIV), adenovirus (ADV) and cytomegalovirus (CMV) by indirect enzyme-linked immunosorbent assay (ELISA). Supernatant INF-γ and IL-4 cultured with PHA were determined. Results The positive rates of IgM in patients with acute exacerbations of COPD compared with those in patients with stable COPD and healthy subjects were significantly different(P <0.001 ). The level of INF-γ, and IL-4 in patients with acute exacerbations of COPD and stable COPD compared with normal control group;the level of INF-γ (242±43) and the of IL-4(42±9) in pa-tients with acute exacerbations of COPD was not significantly different as compared with the level of INF-γ( 198±32) and the level of IL-4(56±11 ) in patients with stable COPD (P<0.05), but the level of INF-γ was increased (90±18)and IL-4 was decreased (141±24) in control group (P<0.05). Compared with those in IgM negative group, the levels of INF-γ were significantly higher and the levels of IL-4 were significantly lowerIL-4 in lgM positive group( P<0.01 ). Conclusion Virus infection is a rather important factor in acute exacerbations of COPD, and COPD is characterized by a predominance of Th1-type response,and there is more significant Th1 response predomi-nance in COPD with virus infection.

Objective To identify human leucocyte antigen (HLA)-A* 0201-restricted hepatitis C virus (HCV)-cytotoxic T lymphocyte (CTL) epitopes. Methods Based on the prediction results of RANKpep and SYFPEITHI prediction programs, six candidate CTL epitopes were selected and synthesized. The affinity of candidate CTL epitopes to HLA-A* 0201 molecules of T2 cells was explored. Subsequently, enzyme-linked immunosorbent spot (ELISPOT) assay and intracellular cytokine staining (ICS) were utilized to determine whether candidate CTL epitopes could induce the recall positive response in peripheral blood mononuclear cells (PBMC) of HLA-A* 0201 positive HCV-1b-infected patients. Results Among six candidate CTL epitopes, peptides C_181(LLSCLTTPV) and NS2_172 (VLQAGLIRV) had high affinity to HLA-A* 0201 molecules. Moreover, the affinity was proportional to the concentration of peptide. Furthermore, among ten HLA-A* 0201 positive HCV-1b-infected patients, the frequencies of C_181 and NS2_172-specific interferon (IFN)-γ-producing cells were 0-19 spots forming cells (SFC)/1 × 105 PBMC and 0-20 SFC/1 × 105 PBMC, respectively.The percentages of C_ 181 and NS2_172-specific IFN-γ+ CD8+ T lymphocytes in total CD8+ T lymphocytes were 0.006%-0.065% and 0.005%-0.080%, respectively. Conclusion Peptides C_181 (LLSCLTTPV) and NS2_172 (VLQAGLIRV) are identified as novel HLA-A* 0201-restricted HCV-CTL epitopes.

Objective To evaluate the preoperative liver function and prognosis of laparoscopic cholecystectomy (LC) in patients with cirrhosis,using the Child-Turcotte-Pugh classification and the model for end-stage liver disease(MELD) score.Methods From January 2009 to June 2013,973 patients who were admitted to the Department of General Surgery of our hospital and the HuiZhou Municipal Central Hosptial were studied.Of the 373 patients with cirrhosis,38 patients were excluded because of Child C,MELD ＞ 30,or laparotomy.The remaining 335 patients who received laparoscopic cholecystectomy were randomly divided into two groups The Child grade and MELD score were retrospectively analyzed.Results There was no significant difference in intraoperative hemorrhage between the Child A group [(106 ± 11) ml] and the Child B group [(109 ± 11) ml] (P ＞ 0.05).The R ＜ 14 scores in the MELD group [(58 ± 15) ml] was significantly lower than that in the R≥ 14 group [(120 ± 28) ml] (P ＜ 0.01).There was no significant difference in postoperative complications between the Child group A (10 cases,12％) and the Child group B (17 cases,21％) (P ＞0.05).There was a significantly lower incidence in the R ＜ 14 scores in the MELD group (10 cases,12％) than the R ≥ 14 group (27 cases,33％) (P ＜ 0.05).There was also no significant difference in the hospital stay between the Child A group (9 ± 1) and the Child B group (10 ± 2)(P ＞0.05) ; the R ＜ 14 score of the MELD group (7 ± 1) was significantly less than that of the R≥ 14 group (11 ±2) (P ＜0.01).There was no significant difference in the cost of hospitalization between the Child A group (1.337 ± 0.063) and the Child B group (1.359 ± 0.089) (P ＞ 0.05) ; the R ＜ 14 group (MELD score 1.108 ± 0.123) was significantly less than that of the R ≥ 14 group (1.568-± 0.117)(P ＜ 0.01).Conclusion Compared with the Child-Turcotte-Pugh classification,the MELD score was more scientific,objective and accurate in judging the preoperative liver function.It helped to predict the amount of intraoperative hemorrhage and postoperative morbidity,reduced hospital stay and hospitalization expenses.Therefore,the MELD scoring system more objectively guided the treatment of patients with cholecystitis with cirrhosis.

Objective To detect the positively selected sites in the surface ( S) gene of hepatitis B viruses ( HBVs) from patients with occult HBV infection and to study the molecular mechanism of occult HBV infection.Methods The sequence of S gene from patients with occult HBV infection and reference strains of eight HBV genotypes ( A through H) were downloaded from GenBank and then alignment analysis were performed by using Clustal W software .Phylogenetic trees were constructed by using MEGA 5.05 soft-ware package.PAML4.7 was used to analyze positively selected sites .Results A total of 1286 HBV se-quences from patients with occult infection were searched in GenBank .One hundred and seventy-four com-plete gene sequences encoding surface S protein were screened after alignment analysis and confirmation , of which 13 sequences with nonsense mutation were removed .The likelihood ratio test showed that for both the 161 remained sequences and the 31 reference sequences , the selection models of M2, M3 and M8 were sig-nificantly better than the neutral models of M0, M1 and M7 (2△lnL<55.12, P<0.001).By using Bayes Empirical Bayes (BEB) analysis, 14 positively selected sites (including codon 3, 8, 40, 45, 46, 47, 49, 68, 126, 127, 164, 184, 207 and 210) were detected in the surface gene of HBVs from patients with occult HBV infection, eight of which were located at the immune epitope of HBsAg .However, only 2 positively se-lected sites were identified in reference sequences .Conclusion The long-lasting persistence of HBV in pa-tients with occult HBV infection might be caused by the adaptive evolution of their surface gene in a form of escape mutant under immune suppressive condition .

Objective To study the nutritional status of pregnant women in Shanxi Province before and after the implementation of the new standards of iodized salt content,provide the basis for scientific supplementation of iodine for pregnant women.Methods According to the method of population proportion sampling,30 county-level monitoring sites were selected,a primary school was selected from each county (city,district) by the method of simple random sampling and 40 students in 2011 or 50 students in 2014 aged 8-10 years were selected in each school,direct titration was used to detect salt iodine;at the same time,20 pregnant women were selected from each town where the primary school was located and urinary iodine was determined using arsenic cerium catalytic spectrophotometry (WS/T 107-2006).Results A total of 1 182 and 1 437 salt samples was detected in Shanxi Province in 2011 and 2014,the median of salt iodine was 30.5 and 24.1 mg/kg,respectively,and the difference was statistically significant (H =567.45,P ＜ 0.01);it was 95.41％,80.31％,76.62％ of the coverage rate of iodized salt,qualified rate of iodized salt,qualified iodized salt consumption rate in 2014,respectively;which were compared with those in 2011 (97.63％,97.49％,95.18％),the differences were statistically significant (x2 =9.27,232.40,166.25,P ＜ 0.01).A total of 440 and 630 urinary samples of pregnant women were tested in 2011 and 2014,the median of urinary iodine was 279.6 and 177.1 μg/L,respectively,iodine nutrition of pregnant women was more than adequate in 2011,and iodine nutrition was suitable in 2014.The difference was statistically significant (H =153.89,P ＜ 0.01).The proportion of pregnant women's median of urinary iodine less than 150 μg/L in 2014 [41.11％ (259/ 630)] was significantly higher than that in 2011 [8.18％ (36/440),x2 =140.68,P ＜ 0.01].The constituent ratio of 250 to 500 μg/L was significantly decreased [23.65％ (149/630) vs 54.77％ (241/440),x2 =108.33,P ＜ 0.01).Conclusion It is at a reasonable level of iodine nutrition level of pregnant women in Shanxi after the adjustment of iodized salt content,but the ratio of ＜ 150 μg/L is increasing,which needs to be paid attention to.