Objective To explore relationship between different HBV genotypes and peripheral blood HBV specific and nonspecific CTL of patients with cirrhotic hepatitis B and its significance. Methods HBV genotypes were tested in 91 patients with cirrhotic hepatitis B, differences of HBV specific and nonspecific CTL between patients infected with genotype B and C were compared and its significance was explored. Results In 91 cases of cirrhotic hepatitis B, 55 cases (60.44% ) belong to genotype C, 35 cases (38. 46% ) belong to genotype B, 1 case (1. 1% ) belongs to mixture genotype B and C. In genotype C,27 cases (49.09% ) had positive (HLA)-A2, HBV specific CTL was 0. 18% ±0.03%. In genotype B, 18 cases (51.43% ) had positive HLA-A2, HBV specific CTL was 0. 38% ± 0.04% , higher than that in genotype C,t =5. 01, P <0. 01. Nonspecific CTL: genotype C (11. 87% ± 1. 50% ) ; genotype B( 11. 90%± 1. 51% ), t =0. 14, P ＜0. 05. HBV DNA level; genotype C (6. 01 ± 0. 81) log10 copy/ml, higher than that in genotype B (5.01 ± 0.54) log10 copy/ml, t =5.01, P ＜0.01. ALT; genotype C (251. 13 ± 131. 11) U/L, higher than that in genotype B (121. 25 ± 63. 21) U/L, t =3. 61, P <0. 01. TBil (45. 61± 15.11) μmol/L, higher than that in genotype B (28.11 ±6.25) μmol/L, t = 3.05, P ＜ 0.01. Conclusion Compared with patients infected with genotype B of cirrhotic hepatitis B, HBV specific CTL of patients infected with genotype C was lower, resulting in higher level of HBV DNA and more severe damage of liver function.

Objective To detect the gene expression changes between urethra plates from hypospadias patients and foreskins from non-hypospadias patients by microarray and to investigate the underlying mechanisms of hypospadias.Methods Twelve hypospadias patients,aged 6-12 months (mean,8 months),were enrolled as the test group,including 5 moderate and 7 severe hypospadias patients.Six age-matched patients underwent circumcision were enrolled as controls.Samples from hypospaidas patient's urethra plates during hypospadias repair and samples from the foreskins during circumcision were obtained and processed into Tri-Reagent immediately for RNA extraction.Oligonucleotide expression microarrays were used to detect genes expression changes in tissues from patients with and without hypospadias.This microarray analysis incorporated 22 000 genes.The intensity of all genes present was analyzed by one-way ANOVA (P＜0.01) and Tukey's test.Four estrogen responsive genes,CYR61,CTGF,ATF3 and GADD45β,were tested by RT-PCR in 8 controls,8 moderate hypospadias and 8 severe hypospadias as well.Results Ninty-four genes were detected differentially expressed in hypospadias patients compared with phimosis patients.There were 47 genes upregulated in moderate hypospadias compared with controls(P＜0.01),68 genes up-regulated in severe hypospadias compared with controls(P＜0.001),17 genes up-regulated in severe hypospadias compared with moderate hypospadias(P＞0.05).These genes were involved in different cell functions such as growth regulation and signal transduction.CYR61,CTGF,ATF3 and GADD45β,known to be estrogen responsive or to interact with estrogen receptor were found up-regulated in microarray and the up-regulations were confirmed by RT-PCR.Conclusions The up-regulated genes contribute to the development of hypospadias.Up-regulation of estrogen responsive genes may play important roles in the development of hypospadias.

The teaching of the Fermentation Engineering Experiment in normal university must serve for the basic education,placing students' creative spirit and practical ability in the first place.Therefore,teaching reform of the Fermentation Engineering Experiment under the background of new curriculum reform of basic education should be studied from the curriculum content,teaching methodology,training pattern and as-sessment system,in order to cultivate the normal-university students' research ability,working attitude,crea-tive and teaching ability.

Objective To explore the association of serum HBV DNA level with HBV-specific and nonspecific cytotoxic T lymphocytes (CTL) in patients with HBV-induced hepatic cirrhosis. Methods 120 patients with HBV-induced hepatic cirrhosis who were positive for HBV DNA, HBeAg and human leucocyte antigen (HLA)-A2 were enrolled in this study. The level of HBV DNA was determined by real time fluorescence quantitative polymerase chain reaction (PCR). HBV-specific and nonspecific CTL were detected by flow cytometry. Liver function tests were done in the 120 patients. The 120 patients were divided into group A and B based on their HBV DNA levels. In group A, there were 68 patients with HBV DNA level of 3-4 log10 copy/ml, and in group B, 52 with 5-6 log10 copy/ml. HBV-specific and nonspecific CTL and liver function were compared between the two groups. Results HBV DNA levels were (3. 68 ± 0. 19)and (5.97 ±0. 32) log10 copy/ml in Group A and B respectively with P ＜ 0. 001. HBV-specific CTL was higher in group A (0.33% ±0.04%) than in group B (0.11% ±0.01%) with P ＜0.001. HBV-nonspecific CTL were (11.99% ±1.51% ) and (11.91% ± 1.61% ) in group A and B respectively with P＞ 0. 05. Conclusion The level of serum HBV DNA is related to the levels of HBV-specific CTL in patients with HBV-induced hepatic cirrhosis. Patients with higher HBV DNA had lower levels of HBV-specific CTL,and the damage to liver function was severe because of higher levels of HBV DNA. Patients with lower HBV DNA had higher levels of HBV-specific CTL which predict good anti-viral effect.

Objective To explore psychological stress factors of patients in the ophthalmology department and psychological nursing interventions as well as effect, to promote the rehabilitation of patients. Methods A total of 289 patients from our ophthalmology department was investigated with the self-designed questionnaire. Results Major psychological stress factors of Ophthalmology inpatient were knowing nothing a variety of points for attention(92％), fearing and worrying the disease and surgery(85％), worrying about the level of health care technology of medical staff (84％). Psychological stress of patients released through psychological nursing intervention measures such as cognitive intervention and environmental intervention. 243 patients of fearing and worrying the disease and surgery reduced to 35 before and after intervene, The difference was statistically significant (X2 = 299.84, P ＜ 0. 01). Conclusions We should properly guide and inspire positive mental patients based on patients' mental state, give mental nursing intervention, promote the rehabilitation of eye disease.

BackgroundObstruction of aqueous humor out flow pathway or abnormality of the extracellular matrix( ECM ) of trabecular meshwork cells causes high intraocular pressure. The balance of matrix metalloproteinases (MMPs) and tissue inhibitors of matrix metalloproteinases(TIMPs) is critical for the metabolism of ECM. Interleukin1α(IL-1α) can influence outflow of aqueous humor by regulating MMPs level. Objective This study was to investigate the effect of interleukin-1α on the expression of MMP-2,MMP-3 and TIMP-I in cultured swine trabecular meshwork cells.Methods Swine sclera with trabecular meshwork tissue was isolated from 20 swine eyes and cultured with explant cultured method. Cultured cells were passaged and third generation cells were identified by fibronectin ( FN ) and laminin ( LN ) staining. After 24 hours of serum starvation, trabecular meshwork cells treated with IL-1α at the concentration of 10 mg/L were regarded as the IL group,and serum-free culture medium used to treat trabecular meshwork cells was regarded as the control group. The expression of MMP-2, MMP-3 and TIMP-1 proteins in trabecular meshwork cells were detected by immunohistochemistry,and the expression of MMP-2 mRNA, MMP-3 mRNA and TIMP-1 mRNA were detected by RT-PCR. The examination results were compared between the two groups. ResultsThe third generation of cells were positive for FN and LM. Compared with the control group, the expression levels of MMP-3 and TIMP-1 proteins(A value) in trabecular meshwork cells were significantly higher in the IL group than the control group(t=-7. 694,t =-5. 199,P＜0. 05) ,but no obvious difference was found in the expression of MMP-2 between the two groups( t=-2. 365, P＞0.05 ). The higher expression levels in MMP-3 mRNA and TIMP-1 mRNA (A value) in trabecular meshwork cells were seen in comparison with the control group (t =-3. 025,t=-1. 921 ,P＜0. 05). However,similar results were found in the expression of MMP-2 mRNA between the two groups(t =- 1. 173, P＞0.05 ). ConclusionsThe overexpression of MMP-3 and TIMP-1 proteins and their mRNA leads to the imbalance of MMP-3/TIMP-1 and promotes the decomposition of ECM in the trabecular meshwork, and therefore increases aqueous outflow.

OBJECTIVETo study the roles of stromal interaction molecule 2 (STIM2) and transient receptor potential canonical 3 (TRPC3) in extracellular Ca(2+)-sensing receptor (CaR)-induced extracellular Ca2+ influx and the production of nitric oxide (NO) in human umbilical vein endothelial cells (HUVEC).

METHODS(1) The interaction of STIM2 and TRPC3 was determined using the immunofluorescence technique. (2) The expressions of STIM2 and TRPC3 genes were silenced in HUVEC by transfection constructed STIM2 and TRPC3 RNA interference plasmids. The interference efficiency of STIM2, TRPC3 protein and mRNA levels were determined by Western blot and real time RT-PCR, respectively. (3) The second to fifth passage of HUVEC were divided into: STIM2-002 short hairpin RNA (STIM2-002 shRNA ) + spermine + Ca2+ group and TRPC3-004 short hairpin RNA (TRPC3-004 shRNA ) + spermine + Ca2+ group; control group (spermine + Ca2+ group) and vehicle+ spermine + Ca2+ group. The four groups of cells were incubated with CaR agonist spermine, the intracellular Ca2+ concentration ([Ca2+]i) was detected using the fluorescence Ca2+ indicator Fura-2/AM, and the production of NO was determined by DAF-FM (NO fluorescent probe) of each group in HUVEC.

RESULTS(1) Immunofluorescence technique results showed that STIM2 and TRPC3 proteinswere present in the cytoplasm of HUVEC. (2) The results of transfection constructed STIM2 and TRPC3 RNA interference plasmids demonstrated that shRNA targeted to the STIM2 and TRPC3 genes decreased STIM2 and TRPC3 mRNA levels by 88.2% and 74.0%, respectively (P < 0.05), simultaneously, the STIM2 and TRPC3 protein levels were decreased by 79.9% and 71.8%, respectively (P < 0.05). (3) Compared with spermine + Ca2+ group, the [Ca2+]i and the net NO fluorescence intensity of spermine + Ca(2+) + ShSTIM2-002 group, spermine + Ca(2+) + ShTRPC3-004 group and spermine + Ca2+ Vehicle group were not changed (P > 0.05).

CONCLUSIONSTIM2 and TRPC3 do not participate in CaR-mediated Ca2+ influx and NO production individually.

Calcium ; metabolism ; Cell Adhesion Molecules ; physiology ; Cells, Cultured ; Human Umbilical Vein Endothelial Cells ; physiology ; Humans ; Nitric Oxide ; metabolism ; Stromal Interaction Molecule 2 ; TRPC Cation Channels ; physiology

OBJECTIVETo establish an analytical method for the fingerprint of triterpenoid constituents of Poria by HPLC and compare the fingerprints of different medicinal parts of Poria in order to provide basis for controlling Poria quality.

METHODThe HPLC chromatographic conditions were Waters Symmetry C18 column (4.6 mm x 250 mm, 5 μm), 0.1% phosphoric acid (A) and acetonitrile (B) as gradient mobile phases, flow rate being 1.0 mL x min(-1), column temperature at 30 degrees C, The detection wavelength was set at 210 nm; The cluster analysis was carried on by SPSS 15.0.

RESULTThe HPLC fingerprints of triterpenoid constituents of Poria were set up. There were 16 common peaks in different medicinal parts. The results of method validation met technical requirement of fingerprints; Triterpenoid constituents in White Poria and Poria cum Radix Pini were different from Poria. The content of pachymic acid was the highest in Poria. The effect of habitat on the quality was no obvious difference.

CONCLUSIONThe method is stable, reliable, reproducible, and can be used as an effective means of Poria quality evaluation.

Chromatography, High Pressure Liquid ; methods ; Cluster Analysis ; Poria ; chemistry ; Triterpenes ; analysis

Objective :To explore related clinical manifestations ,ECG characteristics and therapeutic strategy of ven-tricular arrhythmias induced by secondary hypokalemia .Methods :Clinical data of 184 patients ,who were diagnosed as ventricular arrhythmias induced by secondary hypokalemia in our hospital from Jan 2013 to Nov 2015 ,were retro-spectively analyzed .ECG characteristics and clinical manifestations were analyzed ,blood potassium concentration and incidence of ventricular arrhythmia were compared between before and after treatment .According to blood po-tassium concentration ,patients were divided into mild hypokalemia group (n＝91 ,3.01～3.5mmol/L ,mild group) , medium hypokalemia group (n＝78 ,2.51～3.00mmol/L ,medium group) and severe hypokalemia group (n＝15 ,＜2.5mmol/L ,severe group).ECG condition was compared among these groups .Results : Main clinical manifesta-tions included palpitations ,chest tightness ,fatigue ,hypotension and short of breath etc .,and severe patients can present as coma ,intestinal paralysis and syncope etc .Among ECG manifestations ,premature ventricular systole was the most frequent [75 (40.8%)] ,and the second was frequent premature atrial systole [69 (37.5%)].Compared with mild group ,there were significant rise in percentage of positive ECG changes (50.5% vs.79.5%,100%) in medium group and severe group (P＝0.001 both) ,but there was no significant difference between medium and se-vere group , P＝0.120. Compared with before treatment ,after treatment ,there was significant rise in blood potas-sium concentration [ (2.5 ± 1.1) mmol/L vs.(4.4 ± 1.7) mmol/L] ,and significant reductions in percentages of ventricular premature contraction (86.96% vs .25.00%) and persistent ventricular tachycardia (8.70% vs.0) in these patients ,P＝0.001 all .Conclusion :Along with blood potassium level reduces ,percentage of patients with ven-tricular arrhythmias induced by secondary hypokalemia and abnormal ECG rises .Timely ECG and blood potassium examinations should be performed in these patients for timely treatment .