Seventy-two strains of endophytic fungi were isolated from the healthy bark, branches and leaves of Cephalotaxus hainanensis L.. Sixty-eight of them were morphologically classified into Fungi Imperfecti, thirty-three sporulated were identified to five genera. For those did not sporulate, one was identified to Rhizoctonia sp., the rest were tentatively classified into Mycelia Sterilia. Four were identified to Basidiomycetes. The result indicated the endophytic fungi of C. hainanensis show a degree of tissue specificity. There were significant differences about the quantity, genera and composition between the fungi isolated from bark and those from branches and leaves.

Objective:To investigate the expression of chemokine receptor CXCR4 in hepatocellular carcinoma tissues,hepatocellular carcinoma cell line-MHCC97,human umbilical vein endothelial cells(HUVECs)and the ascites level of CXCL12,ligand of CXCR4,so as to lay a foundation for studying the role of CXCR4 in the metastasis of hepatocellular carcinoma.Methods:The expression of CXCR4 mRNA and protein was examined by RT-PCR and Western blotting in 21 specimens of hepatocellular carcinoma tissues,MHCC97 cells,HUVECs,and 17 specimens of normal hepatic tissues.Meanwhile,the levels of CXCL12 in ascitic fluids were assayed by ELISA in 18 hepatic cancer patients.Results:The relative expression values of CXCR4 mRNA in hepatocellular carcinoma tissues,MHCC97 cells,and HVECs were 2.21?1.09,2.14?1.15 and 1.72?1.20,respectively;and those of CXCR4 protein were 1.51?0.12,1.76?0.25,and 1.89?0.24,respectively;and those of CXCR4 protein were 1.51?0.12,1.76?0.25,and 1.89?0.24,respectively.CXCR4 mRNA and protein were not detected in normal hepatic tissues.ELISA results showed that the 18 hepatocellular carcinoma samples had a CXCL12 concentration range of 783-8 364 pg/ml(median value 6 871 pg/ml)in ascitic fluids.Conclusion:CXCR4 is highly expressed in the hepatocellular carcinoma tissues and cells,which is not associated with the clinical staging of the cancer.The elevated CXCL12 level in the ascitic fluid of cancer patients indicate that CXCR4 may play an important role in the metastasis of hepatocellular carcinoma.

Objective To observe effect of lidocaine pretreatment to malondialdehyde(MDA) and endothelin(ET) of patient ac-cepeted brain tumor removing and discuss the optimized pretreatment time .Methods 60 brain tumor patients in the hospital from March 2009 to September 2011 .according to the different pretreatment time ,the patients were randomly divided into five groups :group A(preoperative 48 h) ,group B(preoperative 24 h) ,group C(preoperative 12 h) ,group D(0 h or anesthesia induced) ,group E (control group) and group F(blank control group) ,10 cases in each group .Group A ,B ,C ,D with 1% lidocaine 1 .5 mg/kg intrave-nous pretreatment on schedule ,then induced conventional anesthesia ;group E were supplemented with 1% lidocaine 2 .5 mg · kg -1 · h-1 intravenous injection after anesthesia induction ;group F was performed routine program without lidocaine .The spontaneous breathing time ,awake time and tracheal extubation time was recorded ,while NIHSS score for evaluation of neural function defect was applied ,and peripheral serum level of MDA and ET was detected by colorimetric technique and radio-immunity .Results In group C ,the spontaneous breathing time ,awake time and tracheal extubation time were shorter than other groups ,but the difference had no statistically significant(P>0 .05) .There was no significant difference among each group in the aspect of NIHSS score 1 day before surgery(P>0 .05) ,after 14 days of operation ,NIHSS of group C was statistically lower than that of group E and group F (P<0 .05) .Before anesthesia induction ,there was no significant difference among groups (P> 0 .05) .MDA and ET content in group C was significantly lower than those in other groups after surgery (P<0 .05) .Conclusion Lidocaine given 12 h before cere-bral ischemia has varying degree protection against cerebral ischemia-reperfusion injury .The protection has relation with the de-crease of MDA and ET content .

This study aims to develop a method for determination of beta-elemene, curcumol, germacrone and neocurdione in the volatile oil of Curcuma phaeocaulis, and to provide the basis of the quality control method for the volatile oil of C. phaeocaulis and the related preparations. Based on GC-MS, the 4 main compounds were simultaneously determined, with the internal standard n-tridecane. The Agilent 19091S-433 column (0.25 microm x 250 microm x 30 m) was adopted at the temperature of 250 degrees C, the programmed temperature method (60 degrees C for 1 min, 5 degrees C x min x to 110 degrees C for 5 min, 1 degrees C x min(-1) to 140 degrees C, 5 degrees C x min(-1) to 160 degrees C, 10 degrees C x min(-1) to 240 degrees C) was used. Helium gas was used as the carrier gas at a constant flow rat of 1 mL x min(-1), with an injection volume of 1 RL. Mass spectra were taken at 70 eV; the ion-source temperature was 200 degrees C. The relation time and character acteristic ions for each target compound were determined by full scan mode and SIM, and m/z 85.1, 93.1, 121.1, 107.1 and 180.1 were the detection ions of n-tridecane, beta-elemene, curcumol, germacrone and neocurdione. As a result, beta-elemene, curcumol, germacrone and neocurdione were all detected with good separation. They were all in a good linear relationship within each concentration scope. The average recovery rates were in the range of 98.2%-101%. So, the method can be used to control the quality of the volatile of C. phaeocaulis Val. and the preparations related.
Acetic Acid ; chemistry ; Curcuma ; chemistry ; Drugs, Chinese Herbal ; analysis ; isolation & purification ; Gas Chromatography-Mass Spectrometry ; methods ; Oils, Volatile ; chemistry ; isolation & purification ; Plant Oils ; chemistry ; isolation & purification ; Sesquiterpenes ; analysis ; isolation & purification ; Sesquiterpenes, Germacrane ; analysis ; isolation & purification

The change of the effective components (liquiritin, glycyrrhizic acid, liquiritigenin, isoliquiritigenin) contents of Glycyrrhizae Radix et Rhizoma (GRR) before and after compatibilities in Sini decoction was studied in this paper. Taking single GRR decoction, GRR-Aconiti Lateralis Radix Praeparata (ALRP) decoction, GRR-Zingiberis Rhizoma (ZR) decoction and Sini decoction as test samples, the contents changing of the four effective components of GRR were measured by HPLC. The results showed that the contents of the four effective components of GRR in the single GRR decoction was higher than that in other samples, and the sequence was single GRR decoction > GRR-ZR decoction > GRR-ALRP decoction > Sini decoction. The contents of liquiritin were 11.18, 9.89, 9.67, 9.17 mg · g(-1); the contents of glycyrrhizic acid were 20.76, 15.58, 11.30, 8.52 mg · g(-1); the contents of liquiritigenin were 0.66, 0.57, 0.45, 0.24 mg · g(-1); the contents of isoliquiritigenin were 0.14, 0.07, 0.03, 0.01 mg · g(-1). Therefore, the effective components of GRR decreased obviously after GRR compatibility with ZR providing scientific basis for GRR relieving the strong nature of ZR. The effective components of GRR decreased sharply after GRR compatibility with ALRP providing scientific support for the material foundation research of GRR reducing the toxicity of ALRP. The effective components of GRR decreased further in Sini decoction indicating that the three medicines in Sini decoction were interactional, which reflecting the scientific connotation of the mutual-restraint/mutual-detoxication, mutual-promotion/mutual-assistance compatibilities in Sini decoction.
Chromatography, High Pressure Liquid ; Drug Compounding ; methods ; Drugs, Chinese Herbal ; analysis ; Glycyrrhiza ; chemistry ; Rhizome ; chemistry

Using the concepts and methods of epigenetics and metabolomics, to investigate the overall action molecular mechanism of Chrysanthemi indici C (CIC), the anti-hepatitis B virus (HBV) active extracts from Flos chrysanthemi indici. The inhibitory effects of CIC on proliferation and hepatitis B surface antigen (HBsAg), hepatitis B envelope antigen (HBeAg) and HBV-DNA of HepG2.2.15 cells were detected by CCK-8 and antigen kit. The DNA methyltransferases (DNMTs)/ten-eleven-translocation-2 (TET2) equilibrium was detected by ELISA. Illumina 850K methylation chip, pyrosequencing and qPCR were used to determine the action pathway and target of CIC by GO and KEGG analysis. Cell metabolites were extracted with 80% methanol, and the changes of differential metabolites, differential metabolic pathways and cell microenvironment were detected by LC-MS and other metabolomics methods. The results showed that CIC could inhibit the proliferation, HBsAg, HBeAg and HBV-DNA of HepG2.2.15 cells obviously, down-regulate DNA methyltransferase 1 (DNMT1), DNA methyltransferase 3a (DNMT3a) and DNA methyltransferase 3b (DNMT3b), up-regulate TET2, and restore the balance of DNMTs/TET2. The action targets of CIC were phospholipase C gamma 2 (PLCG2), phosphoinositide-3-kinase regulatory subunit 3 (PIK3R3), 1-acylglycerol-3-phosphate O-acyltransferase 2 (AGPAT2), 5-hydroxytryptamine receptor 2B (HTR2B), nerve growth factor (NGF), mainly involved in lipid metabolism, inflammation mediated regulation of transient receptor potential (TRP), phospholipase D signaling and advanced glycation end product-receptor for AGE (AGE-RAGE) signaling in diabetic complications pathways. CIC could significantly affect fatty acid metabolism and had great influence on phenolic acid, alkaloid and lipid metabolites in cell microenvironment. These results suggest that the action mechanism of CIC may be the synergistic action of multiple pathways and multiple targets, including related inflammatory pathways, immune pathways and lipid metabolism, through regulating epigenetic expression balance and restoring the balance of cell microenvironment.

Objective:To observe the effect of point-moxibustion on gastrointestinal motility,mRNA and protein expressions of ghrelin and growth hormone secretagogue receptor 1a (GHSR-1a) in lateral septal nucleus of rats with diabetic gastroparesis (DGP),and to investigate the central regulatory mechanism of DGP treatment with point-moxibustion.Methods:Forty SPF-grade Sprague-Dawley (SD) rats were randomly divided into a blank group,a model group,an electroacupuncture (EA) group and a point-moxibustion group,with 10 rats in each group.A DGP rat model was established by intraperitoneal injection of 2％ streptozotocin (STZ) with 8-week irregular high-sugar and high-fat diet in the model group,the EA group and the point-moxibustion group;and rats in the blank group were injected intraperitoneally with 0.1 mmoL/L (pH 4.5) citric acid-sodium citrate buffer with 8-week normal diet.Eight weeks later,rats in the EA group were treated by EA at Zusanli (ST 36),Liangmen (ST 21) and Sanyinjiao (SP 6);while rats in the point-moxibustion group were treated by point-moxibustion at Zusanli (ST 36),Liangmen (ST 21) and Sanyinjiao (SP 6) for successive 15 d.Rats in the blank group and the model group were fixed as the control without intervention.After treatment,intestinal propulsion rate and gastric emptying rate were measured.The mRNA and protein expressions of ghrelin and GHSR-1a in the lateral septal nucleus were detected by real-time polymerase chain reaction (RT-PCR) and Western blot (WB).Results:Compared with the blank group,the intestinal propulsion rate and gastric emptying rate of the model group were significantly lower (both P＜0.01);compared with the model group,the intestinal propulsion rate and gastric emptying rate of the EA group and the point-moxibustion group increased significantly (all P＜0.05).The mRNA and protein expressions of ghrelin and GHSR-1a were lower in the model group than those in the blank group (all P＜0.01).The mRNA and protein expressions of ghrelin and GHSR-1a were significantly higher in the EA group and the point-moxibustion group than those in the model group (all P＜0.05).There were no statistically significant differences between the EA group and the point-moxibustion group (all P＞0.05).Conclusion:Point-moxibustion at Zusanli (ST 36),Liangmen (ST 21) and Sanyinjiao (SP 6) can increase the intestinal propulsion rate and gastric emptying rate of DGP rats,and promote the mRNA and protein expressions of ghrelin and GHSR-1a in the central nervous system.The mechanism may be related to the activation of ghrelin pathway in hypothalamic arcuate nucleus-lateral septal nucleus.

OBJECTIVETo evaluate the effect of surgical timing of intertrochanteric fracture on 1-year postoperative mortality in elderly and analyze factors that influence 1-year postoperative mortality.

METHODSA retrospective search had been conducted to analyze 350 patients of intertrochanteric fracture from December 2012 to December 2014. The studied patients were divided into three groups such as the group of <72 h and the group of 72-96 h and the group of >96 h, which depended on the time of injury to surgery. The postoperative mortality was compared among the groups and causes of death in patients were evaluated after surgery. Multiple logistic regression was then used to distinguish independent effects on mortality.

RESULTSThe one-year postoperative mortality of three groups(<72h, 72-96 h, >96 h)were significantly different and the rate of death was 1.8%, 8.1%, 10.3%. The one-year postoperative mortality of the group of <72 h was inferior to the group of 72-96 h and >96 h respectively(<0.05). Age, preoperative coexist disease and surgical timing were dependent risk factors to one-year postoperative mortality(<0.05, OR>1).

CONCLUSIONSThe early surgery can diminish one year postoperative mortality, the patient who is older and has too many preoperative coexist diseases and is delayed to surgery has higher risk to die in one year after surgery.

OBJECTIVETo investigate the changes in angiotensinogen (AGT), angiotensin II type 1 receptor (AT(1)R), endothelial nitric oxide synthase (eNOS) mRNA and protein expressions and nitric oxide (NO) content in the rat glomeruli in response to leptin stimulation.

METHODSThe glomeruli isolated from male SD rats were stimulated with 3 nmol/L leptin for 2 h. Real-time PCR and Western blotting were performed to analyze the mRNA and protein expressions of AGT, AT(1)R and eNOS in the glomeruli, and nitrite concentration in the glomeruli was measured by nitrate reductase assay.

RESULTSIn comparison with the control group, exposure to leptin increased the mRNA levels of AGT, ATR(1) and eNOS in the isolated glomeruli by 2.69-/+0.17, 3.77-/+0.16 and 2.56-/+0.29 folds (P=0.024, 0.018 and 0.044), and their protein levels by 2.06-/+0.10, 2.67-/+0.08 and 1.61-/+0.13 folds (P=0.021, 0.015 and 0.032), respectively. The NO production in the glomeruli was also increased by 2.77-/+0.14 folds (P=0.000) following leptin exposure.

CONCLUSIONLeptin exposure of isolated rat glomeruli directly causes activation of the internal renal renin-angiotensin system and enhanced NO production, suggesting that leptin plays a role in the pathogenesis of maladaptation in renal hemodynamics in obesity.

Animals ; Gene Expression Regulation ; drug effects ; Kidney Glomerulus ; drug effects ; metabolism ; Leptin ; pharmacology ; Male ; Nitric Oxide ; biosynthesis ; Nitric Oxide Synthase Type III ; genetics ; metabolism ; RNA, Messenger ; genetics ; metabolism ; Rats ; Rats, Sprague-Dawley ; Receptor, Angiotensin, Type 1 ; genetics ; metabolism ; Renin-Angiotensin System ; drug effects

OBJECTIVEThe prevalence of obesity and of type 2 diabetes mellitus in children have increased in the Chinese population over the past two decades, and thus diabetes prevention has become a major concern of public health agencies. Identification of individuals at risk for diabetes is an essential first step in designing and implementing intervention programs. Insulin resistance is the hallmark of the pathophysiology of type 2 diabetes mellitus. Subjects with hyperinsulinemia and impaired glucose tolerance are well accepted as being at high risk for diabetes. Acanthosis nigricans (AN) has been proposed as a reliable marker of hyperinsulinemia, but its utility for predicting hyperinsulinemia has not been systematically evaluated in obese children. In order to further explore the relationship between obese childhood with benign acanthosis nigricans and insulin resistance and type 2 diabetes mellitus, we examined 19 obese children with benign acanthosis nigricans.

METHODSNineteen of seventy six obese children (25%) with BMI over 25 enrolled in the Children' Hospital of Zhejiang University School of Medicine fromJune 1st to September 1st in 2003 were studied. Skin biopsies were performed in these 19 obese children with acanthosis nigricans for final diagnosis. Levels of glucose, insulin, and glucose/insulin ratio were measured on fasting blood specimens and anthropometric parameters including waist/hip ratio, fat mass, body fat percentage and body mass index were examined. Oral glucose tolerance tests were also performed in these 19 children with benign acanthosis nigricans.

RESULTSAnthropometric parameters including waist/hip ratio, fat mass, body fat percentage and body mass index as well as fasting insulin level in acanthosis nigricans group were significantly higher than that of healthy controls (P < 0.01). Fasting glucose to insulin ratio (FGIR) of these 19 obese children with benign acanthosis nigricans was 4.27 +/- 0.53, indicating apparent insulin resistance. One of them was diagnosed as type 2 diabetes mellitus and ten of them showed impaired oral glucose tolerance.

CONCLUSIONChildhood benign acanthosis nigricans is tightly associated with obesity, hyperinsulinemia, insuline resistance and type 2 diabetes mellitus, and may be used as a reliable index of insulin resistance.

Acanthosis Nigricans ; complications ; physiopathology ; Child ; Diabetes Mellitus, Type 2 ; complications ; physiopathology ; Humans ; Hyperinsulinism ; complications ; physiopathology ; Insulin Resistance ; Obesity ; complications ; physiopathology