Molecular Characteristics and Potent Immunomodulatory Activity of Fasciola hepatica Cystatin

Cystatin, a cysteine protease inhibitor found in many parasites, plays important roles in immune evasion. This study analyzed the molecular characteristics of a cystatin from Fasciola hepatica (FhCystatin) and expressed recombinant FhCystatin (rFhcystatin) to investigate the immune modulatory effects on lipopolysaccharide-induced proliferation, migration, cytokine secretion, nitric oxide (NO) production, and apoptosis in mouse macrophages. The FhCystatin gene encoded 116 amino acids and contained a conserved cystatin-like domain. rFhCystatin significantly inhibited the activity of cathepsin B. rFhCystatin bound to the surface of mouse RAW264.7 cells, significantly inhibited cell proliferation and promoted apoptosis. Moreover, rFhCystatin inhibited the expression of cellular nitric oxide, interleukin-6, and tumor necrosis factor-α, and promoted the expression of transforming growth factor-β and interleukin-10. These results showed that FhCystatin played an important role in regulating the activity of mouse macrophages. Our findings provide new insights into mechanisms underlying the immune evasion and contribute to the exploration of potential targets for the development of new drug to control F. hepatica infection.

Molecular detection and genetic diversity of bovine papillomavirus in dairy cows in Xinjiang, China

Background@#Bovine papillomatosis is a type of proliferative tumor disease of skin and mucosae caused by bovine papillomavirus (BPV). As a transboundary and emerging disease in cattle, it poses a potential threat to the dairy industry. @*Objectives@#The aim of this study is to detect and clarify the genetic diversity of BPV circulating in dairy cows in Xinjiang, China. @*Methods@#122 papilloma skin lesions from 8 intensive dairy farms located in different regions of Xinjiang, China were detected by polymerase chain reaction. The genetic evolution relationships of various types of BPVs were analyzed by examining this phylogenetic tree. @*Results@#Ten genotypes of BPV (BPV1, BPV2, BPV3, BPV6, BPV7, BPV8, BPV10, BPV11, BPV13, and BPV14) were detected and identified in dairy cows. These were the first reported detections of BPV13 and BPV14 in Xinjiang, Mixed infections were detected, and there were geographical differences in the distribution of the BPV genotypes. Notably, the BPV infection rate among young cattle (< 1-year-old) developed from the same supply of frozen sperm was higher than that of the other young cows naturally raised under the same environmental conditions. @*Conclusions@#Genotyping based on the L1 gene of BPV showed that BPVs circulating in Xinjiang China displayed substantial genetic diversity. This study provided valuable data at the molecular epidemiology level, which is conducive to developing deep insights into the genetic diversity and pathogenic characteristics of BPVs in dairy cows.

Molecular detection and genetic diversity of bovine papillomavirus in dairy cows in Xinjiang, China

Background@#Bovine papillomatosis is a type of proliferative tumor disease of skin and mucosae caused by bovine papillomavirus (BPV). As a transboundary and emerging disease in cattle, it poses a potential threat to the dairy industry. @*Objectives@#The aim of this study is to detect and clarify the genetic diversity of BPV circulating in dairy cows in Xinjiang, China. @*Methods@#122 papilloma skin lesions from 8 intensive dairy farms located in different regions of Xinjiang, China were detected by polymerase chain reaction. The genetic evolution relationships of various types of BPVs were analyzed by examining this phylogenetic tree. @*Results@#Ten genotypes of BPV (BPV1, BPV2, BPV3, BPV6, BPV7, BPV8, BPV10, BPV11, BPV13, and BPV14) were detected and identified in dairy cows. These were the first reported detections of BPV13 and BPV14 in Xinjiang, Mixed infections were detected, and there were geographical differences in the distribution of the BPV genotypes. Notably, the BPV infection rate among young cattle (< 1-year-old) developed from the same supply of frozen sperm was higher than that of the other young cows naturally raised under the same environmental conditions. @*Conclusions@#Genotyping based on the L1 gene of BPV showed that BPVs circulating in Xinjiang China displayed substantial genetic diversity. This study provided valuable data at the molecular epidemiology level, which is conducive to developing deep insights into the genetic diversity and pathogenic characteristics of BPVs in dairy cows.

Characteristic profiles of biofilm, enterotoxins and virulence of Staphylococcus aureus isolates from dairy cows in Xinjiang Province, China

As an important zoonotic pathogen, Staphylococcus aureus has led to serious mastitis and endometritis in infected dairy cows. In this study, a total of 164 strains of S. aureus were isolated from dairy cows in Xinjiang Province, China, and subjected to assays to determine drug susceptibility and biofilm (BF) formation ability. Enterotoxin-related genes were detected, and the transcription levels of genes related to BF formation were determined by using reverse transcription-quantitative polymerase chain reaction. Moreover, the pathogenicity of isolates with different BF formation abilities was determined by measuring their hemolysis activity, half lethal dose (LD₅₀) and organ bacterial load. The results showed that 86.0% of S. aureus isolates could form BF. Among them, 42.1% of the strains had weak BF formation ability, and most strains with a strong BF formation ability were ica gene carriers. The S. aureus isolates displayed multidrug resistance and their drug resistance was positively correlated with their BF formation ability. Moreover, 96.3% of the S. aureus isolates carried enterotoxin genes. Among them, the detection rates of the novel enterotoxin genes were higher than those of conventional enterotoxin genes. Furthermore, isolates with a strong BF formation ability had higher LD50 but lower hemolysis ability and organ bacterial load than those of the isolates with weak or no BF ability. However, isolates without BF ability produced more severe pathological changes than those of isolates with strong BF formation ability. These findings suggest that higher BF ability and presence of novel enterotoxin genes are important characteristics of S. aureus isolates from dairy cows in Xinjiang Province, China, and such isolates may pose potential threats to food safety.

Preparation of peptide mimotope-based diagnostic antigen of Epstein-Barr virus infection / 中华实验和临床病毒学杂志

Objective@#To prepare peptide minotope-based recombinant diagnostic antigen of Epstein-Barr virus (EBV) infection and evaluate its antigenicity preliminarily.@*Methods@#With Trx at the N-terminal and His tag at the C-terminal, the peptide minotope of EBV (GP125, F1, A2, A3C2) was expressed in Escherichia coli and purified by affinity and anion exchange chromatography (designated 'H58’); based on antigenicity of H58 identified by Western blotting (WB), we constructed and evaluated a novel early diagnostic ELISA for EBV infection.@*Results@#The soluble H58 protein with high concentration (2.8 mg/ml) and purity (99.01) was obtained; WB analysis found that there was an obvious band (28 ×103) on the NC membrane, using H58 anti-Trx monoclonal antibody or acute-phase sera of EBV infection as the first antibody. With the novel ELISA, 50 positive sera of EBV infection and 50 negative sera were detected, displaying that the grouping of OD value of positive serum (95%CI: 1.233-1.489) and negative serum (95%CI: 0.113-0.159) was different (P<0.05) with the sensitivity 98.0%, specificity 96.0% and kappa value 0.940.@*Conclusions@#By E. coli expression and affinity and ion exchange chromatography purification, the peptide minotope-based recombinant diagnostic antigen of EBV infection was obtained with excellent antigenicity, which could be applied for serological detection of EBV infection.

Clinical effect of lamotrigine combined with small dose of valproic acid in the treatment of newly diagnosed epilepsy / 中国基层医药

Objective To explore the clinical effect of lamotrigine combined with small dose of valproic acid in the treatment of newly diagnosed epilepsy .Methods 90 patients with newly diagnosed epilepsy were selected ,and they were randomly divided into control group and observation group according to the digital table ,45 cases in each group.The control group was only given small dose valproic acid treatment ,the observation group was given lamotrigine based on the control group .The curative effect ,clinical indicators ,and the change of cognitive function were compared between two groups .Results The total effective rate of the control group was 82.22％,which was significantly lower than 91.11％of the observation group (χ2 =5.36,P<0.05).After treatment,the attack frequency,duration,the attack of epilepsy discharge,involving lead number of the control group were (1.29 ±0.55)times per year,(3.36 ± 0.63)min,(11.69 ±1.26)180s/time,(5.69 ±1.52)180s/time,which of the observation group were (0.51 ± 0.22)times per year,(2.09 ±1.02)min,(7.56 ±1.34)180s/time,(3.54 ±1.48)180s/time,the differences between the two groups were statistically significant (t=4.18,4.28,5.14,4.28,all P<0.05).Compared with before treatment,the MMSE,digit span along the back test B scores after treatment in the two groups were increased ,and the indicators of the observation group after treatment improved more significantly than those in the control group ( t=4.18,4.28,5.14,all P<0.05).Conclusion The therapeutic effect of lamotrigine combined with small dose of valproic acid in the treatment of newly diagnosed epilepsy is significant , which can reduce clinical indicators and improve cognitive function .

Sequence analysis of HBV in primary hepatomas patients infected with HBV / 中华实验和临床病毒学杂志

Objective@#To study the relationship between the development of hepatocellular carcinoma(HCC) and HBV gene characteristics among the HCC patients with hepatitis B virus (HBV) infection.@*Methods@#Some acute and chronic hepatitis B patients were collected as control group and HBV associated HCC patients as HCC group. Serum samples of subjects were tested for HBV serological markers. HBV DNA of those samples had been extracted and nested PCR was used to amplify the sequence of HBV DNA. Furthermore, MEGA 6.0 and Bioedit softwares were used to made phylogenetic trees and analyze the gene mutations.@*Results@#The sequences of S region and BCP/Precore region of HBV were amplified from 86 samples in study group and 39 samples in control group. The prevalence of PreS deletion, A1762T and A1762T/G1764A in HCC group were 39.53%, 74.42% and 72.09% respectively, and in control group were 20.51%, 53.85% and 53.85% respectively. The statistical differences of them were significant. The prevalence of A1762T and A1762T/G1764A in ≥ 50 years group were higher than that of < 50 years group. The prevalence of A1762T, G1764A and A1762T/G1764A of subjects who infected genotype C were higher than those infected genotype B. On the contrary, the prevalence of G1896A of subjects who infected genotype C were lower than that of genotype B. It was found that ≥ 50 years, genotype C and G1896A mutation were independently associated with HCC. The risk for suffer from HCC of ≥50 years group, genotype C group and G1896A group were 9.349, 28.875 and 7.648 times compared with < 50 years group genotype B group and without G1896A mutation group, respectively.@*Conclusions@#The population of ≥50 years or genotype C had a higher prevalence of A1762T, A1762T/G1764A, ≥50years、genotype C、G1896A were independently associated with HCC, as compared with the subjects of the control group.

Risk factors of hemorrhagic transformation after cerebral infarction / 中国医师进修杂志

Objective To investigate the risk factors of hemorrhagic transformation (HT) after cerebral infarction. Methods The clinical and neuroimaging data of 100 patients (gender,age-matched)with cerebral infarction and hemorrhagic transformation from January 2005 to January 2010 were analyzed retrospectively. The following factors were listed for single factor analysis: the history of hypertension, diabetes mellitus,admission blood pressure,blood sugar,infarction size,alcohol,smoking,atrial fibrillation,hypercholesterolemia, combined anticoagulant and antiplatelet aggregation therapy. All the above factors were analyzed by chi-square test or t test, and Logistic regression analysis was used to screen out the related risk factors of HT from the potentially related factors. Results Atrial fibrillation (OR =5.483,95%CI:fibrillation,extensive cerebral infarction,blood sugar,high blood pressure at the beginning of the disease,combined anticoagulant and antiplatelet aggregation therapy are the major risk factors of HT. At the same time, the occurrence of HT is the result of the joint effect of multiple factors and multiple mechanisms.

ANATOMIC STUDY OF MAXIMUM INTENSITY PROJECTION IN MRI FOR MEMBRANOUS LABYRINTH AND INTERNALAUDITORY MEATUS IN 15 CHINESE ADULTS / 解剖学报

Objective The three-dimensional reconstructed images of maximum intensity projection (MIP) for membranous labyrinth and internal auditory meatus in Chinese adults were observed and measured in order to provide anatomic basis for otolosurgery and nerosurgery. Methods Thirty inner ears of 15 volunteers were scanned by using a circular temporal coil and three-dimension fast spin echo sequence with a 1