ObjectiveTo investigate the histological features as well as the factors influencing liver disease progression in Chinese patients with chronic hepatitis C (CHC). MethodsA total of 102 CHC patients who underwent percutaneous liver biopsy between August 2007 and May 2010 were recruited. Age, gender, body mass index (BMI) and transmission route of recruited patients were recorded. Serum levels of alanine transaminase (ALT) and aspartate transaminase (AST), HCV genotypes, HCV viral load and liver histological changes were detected. Statistical analysis was done by t test and Logistic regression. ResultsThe serum levels of ALT and AST in CHC patients with histological activity index (HAI) ≥4 were much higher, while platelet (PLT) counts were lower than those with HAI ＜4(t=2.209, 2. 298 and 2. 565, respectively; all P＜0.05). Likewise, in patients with F≥3, the serum levels of ALT and AST as well as the mean age and the duration of infection were significantly elevated compared with F ＜ 3 group ( t = 3.497, 2. 758, 2. 340 and 2. 570,respectively; all P＜0. 05), while PLT counts were much lower (t = 2. 761, P=0. 007). The unvariate predictors for HAI≥4 were female, ALT＞1 × upper limits of normal (ULN), AST level,F≥3, HCV RNA≥6 lgIU/mL and PLT counts. By mutivariate analysis, the Ishak stage score was the only independent predictor for HAI≥4 (OR 3.098, 95％CI 1.884-5. 092; P＜0.01). Finally,the univariate predictors for F≥3 were age, BMI≥24 kg/m2 , ALT＞1 × ULN, AST level, HAI≥4,PLT counts and duration of infection≥ 15 years. Multivariate analysis revealed that age (OR 1. 074,95％CI 1.006-1. 146; P=0.033), ALT level (OR 1. 035, 95％CI 1.015－1.055; P＜0.01), ASTlevel (OR 0. 969, 95％CI 0. 948－0. 990; P=0. 005), the duration of infection ≥15 years (OR 37. 215, 95％CI 5. 816－238. 127; P＜0.01) and HAI≥4 (OR 1. 939, 95％CI 1. 426－2. 636; P＜0.01) were independent predictors for F≥ 3. ConclusionAge, ALT level, AST level, duration of infection≥15 years, HAI≥4 are independent predictors for liver fibrosis.

This study aimed to examine the preparation of cationic lipid microbubble (CLM), and evaluate its physical and chemical properties and toxicity, measure the gene transfection efficiency by ultrasound triggered microbobble destruction (UTMD) in combination with CLM. The CLM was prepared by the method of the thin film hydration, and its morphology was observed under the electron microscopy at 1st, 3rd, 7th, 10th, and 14th day after preparation, respectively. The size, Zeta potential and stability of CLM were tested. The acute toxicity of CLM was assessed. The green fluorescent protein gene (EGFP) transfection efficiency was evaluated. The experiment grouping was as follows: naked plasmid group (P group), ultrasonic irradiation plus naked plasmid group (P-US group), naked plasmid plus CLM group (P-CLM group), naked plasmid plus ultrasound and CLM group (UTMD group). The expression of EGFP was detected by fluorescent microscopy and flow cytometry. The results showed that CLMs were spherical in shape, with the similar size and good distribution degree under the light and electron microscopies. The size of CLMs was varied from 250.4±88.3 to 399.0±99.8 nm and the Zeta potential of CLMs from 18.80±4.97 to 20.1±3.1 mV. The EGFP expression was the strongest in the UTMD group, followed by the P-CLM group, P-US group and P group. Flow cytometry results were consistent with those of fluorescent microscopy. The transfection efficiency was substantially increased in the P-US group, P-CLM group and UTMD group as compared with that in the P group, almost 7 times, 10 times and 30 times higher than that in the P group respectively. It is suggested that CLMs prepared by the method of thin film hydration are uniform in diameter, and proved non-toxic. UTMD combined with CLM can significantly increase the transfection efficiency of EGFP to targeted cells.

OBJECTIVETo investigate telomerase activity in rabbit bone marrow stromal cells (BMSCs) during their committed differentiation in vitro along neural pathway and the effect of glial cell line-derived neurotrophic factor (GDNF) on the expression of telomerase.

METHODSBMSCs were acquired from rabbit marrow and divided into control group, GDNF (10 ng/ml) group. Cytokine.NSCs medium (prepared by our lab, Patent No. ZL02134314. 4) supplemented with 10 percent fetal bovine serum (FBS) was used to induce BMSCs differentiation along neural pathway. Fluorescent immunocytochemistry was employed to identify the expressions of Nestin, neuron-specific endase (NSE), and gial fibrillary acidic protein (GFAP). The growth curves of the cells and the status of cell cycles were analyzed, respectively. During the differentiation, telomerase activities were detected using the telomeric repeat amplification protocol-enzyme-linked immunosorbent assay (TRAP-ELISA).

RESULTSBMSCs were successfully induced to differentiate along neural pathway and expressed specific markers of fetal neural epithelium, mature neuron and glial cells. Telomerase activities were undetectable in BMSCs during differentiation along neural pathway. Similar changes of cell growth curves, cell cycle status and telomerase expression were observed in the two groups.

CONCLUSIONSRabbit BMSCs do not display telomerase activity during differentiation along neural pathway. GDNF shows little impact on proliferation and telomerase activity of BMSCs.

Animals ; Bone Marrow Cells ; enzymology ; Cell Differentiation ; Glial Cell Line-Derived Neurotrophic Factor ; Immunohistochemistry ; Rabbits ; Stromal Cells ; enzymology ; Telomerase ; metabolism

OBJECTIVETo compare the expressions of ODF1 (outer dense fiber of the sperm tail 1) in ejaculated spermatozoa from normozoospermic and asthenozoospermic men with low sperm motility.

METHODSSemen analyses were performed on the semen samples obtained from normozoospermic (n=20) and asthenozoospermic (n=20) volunteers according to the WHO criteria. To rule out the contamination of germ cells and leucocytes, the human ejaculated spermatozoa were purified by a discontinuous Percoll density gradient centrifugation. RT-PCR and Western blot were used to detect the expressions of ODF1 in the spermatozoa from the two groups.

RESULTSRT-PCR showed that the expression of ODF1 mRNA was significantly lower in the spermatozoa from the asthenozoospermic patients than in those from the normozoospermic men (1.35 +/- 0.25 vs. 2.79 +/- 0.28, P < 0.05). Western blot confirmed the results from RT-PCR and revealed an obviously decreased expression of ODF1 in the spermatozoa of the asthenozoospermic patients, with statistically significant difference from the normozoospermic group (1.44 +/- 0.26 vs. 3.64 +/- 0.34, P < 0.05).

CONCLUSIONThe expression of ODF1 was significantly decreased in the ejaculated spermatozoa of asthenozoospermic men, which might be responsible for low sperm motility.

Asthenozoospermia ; metabolism ; Heat-Shock Proteins ; metabolism ; Humans ; Male ; Sperm Motility ; Spermatozoa ; metabolism

OBJECTIVESPAG9, as a member of the MAPK family, plays an important role in sperm-egg fusion. This study aimed to detect the expression of SPAG9 in human ejaculated spermatozoa.

METHODSDifferent human tissues (as from the muscle, liver, esophagus, lung, stomach, kidney, prostate, uterus, testis and epididymis) and semen samples were obtained from healthy volunteers, and semen analyses were performed according to the WHO criteria. Human ejaculated spermatozoa were purified by discontinuous Percoll density gradient centrifugation to rule out the contamination of germ cells and leucocytes. RT-PCR and indirect immunofluorescence were used to detect the expression of SPAG9 in human spermatozoa.

RESULTSRT-PCR showed that SPAG9 mRNA was expressed in different tissues and human ejaculated spermatozoa. Indirect immunofluorescence studies revealed the location of SPAG9 protein in the equatorial plate and flagella of human spermatozoa.

CONCLUSIONSPAG9 is expressed in ejaculated spermatozoa and may play a role in sperm capacitation and motility.

Adaptor Proteins, Signal Transducing ; genetics ; metabolism ; Fluorescent Antibody Technique, Indirect ; Humans ; Male ; Reverse Transcriptase Polymerase Chain Reaction ; Spermatozoa ; metabolism

Comparative analysis of variable region ORF14/15 genes of white spot syndrome virus (WSSV) genome in Guangxi Penaeus vannamei (P. vannamei) could provide useful information for the evaluation of genetic diversity and genetic evolutionary relationship among WSSV isolates from Guangxi, China and other places. Based on geographical and temporal considerations, 40 WSSV-positive P. vannamei samples were collected during the period between May 2010 and July 2013 from Beihai, Qinzhou, and Fangchenggang, which were the main P. vannamei production areas in Guangxi, and the variable region ORF14/15 genes of the WSSV genome from all infected samples were amplified by PCR and then subjected to cloning and sequence analysis. Pairwise and multiple alignment analysis was then conducted to evaluate the degree of genetic divergence between different strains. The variable region ORF14/15 genes from 25 of 40 WSSV positive samples were successfully cloned and sequenced; among the ORF14/15 genes of 25 WSSV-positive strains, 22 was 619 bp in length and 3 was 620 bp. All the 25 Guangxi strains carried a 5949-bp deletion in the ORF14/15 region relative to TH-96-II, which has the longest nucleotide sequence in this region; the deletion of Guangxi strains occurred in the middle region of ORF14/15 gene, with only 190 bp and 429 bp/ 430 bp at 5' and 3' ends, respectively, which were coincident with WSSV-IN-05-I in deletion length and position. Sixteen of 25 Guangxi strains had completely identical nucleotide sequences in the variable re gion, and the homology between other strains also exceeded 97.9%. There were single nucleotide substi tution, deletion, and insertion in the ORF14/15 region of Guangxi strains compared with other strains in GenBank. In the phylogenetic tree based on WSSV variable region ORF14/15, the Guangxi strains were closely related and formed a separate branch with Indian strain IN-05-I, but far from other strains in GenBank. The ORF14/15 gene of WSSV isolates in cultured P. vannamei in Guangxi has a large deletion in the middle of the variable region, and the Guangxi WSSV strains show no significant spatio-temporal differences; the Guangxi strains are closer in genetics to Indian strain IN-05-I than other strains in GenBank.
Animals ; China ; Cloning, Molecular ; Evolution, Molecular ; Genome, Viral ; genetics ; Genomics ; Penaeidae ; virology ; Phylogeny ; White spot syndrome virus 1 ; genetics

Animals ; Liver Cirrhosis ; metabolism ; prevention & control ; Male ; Rats ; Rats, Inbred Strains ; Thiazolidinediones ; therapeutic use ; Tissue Inhibitor of Metalloproteinase-1 ; metabolism

To investigate the inhibition of cyclosporin A (CsA) on neutrophil adhesion to human umbilical vein endothelial cells (HUVECs, ECV-304) induced by hypoxia/reoxygenation and further explore its mechanism, a 1 h hypoxia/4 h reoxygenation model was reproduced using ECV-304. The adhesion rate of neutrophils to ECV-304 was determined by measuring the activity of endogenous hexosaminidase. The expression of endothelial cell adhesion molecules of E-selectin and ICAM-1 was measured by flow cytometry. The expression of cyclophilin A (CyPA) and the activation of ERK1/2 was compared among experimental groups by Western blot. The content of reactive oxygen species (ROS) was measured by Fenton reaction. After being stimulated with 1 h hypoxia/4 h reoxygenation, ECV-304 showed an enhanced neutrophil adhensiveness in association with an increased surface expression of E-selectin and ICAM-1. In parallel, the content of ROS was also increased. These effects were significantly suppressed by the addition of CsA. Most importantly, the expression of CyPA was significantly increased following 1 h hypoxia/4 h reoxygenation, which was accompanied with an increased activation of ERK1/2. Treatment with CyPA inhibitor CsA and CyPA antisense oligonucleotides significantly inhibited the activation of ERK1/2 and decreased the adhesion of neutrophils to ECV-304. The specific ERK1/2 inhibitor PD98059 caused an inhibition of neutrophil adhesion to hypoxia/reoxygenation-stimulated ECV-304. Our data confirm that CsA inhibits neutrophil adhesion to hypoxia/reoxygenation stimulated ECV-304 by a mechanism involving inhibition of the signal transduction of ROS, CyPA and ERK1/2.
Cell Adhesion ; Cell Hypoxia ; Cells, Cultured ; Cyclophilins ; biosynthesis ; genetics ; Cyclosporine ; pharmacology ; Endothelium, Vascular ; cytology ; Humans ; Intercellular Adhesion Molecule-1 ; biosynthesis ; genetics ; Mitogen-Activated Protein Kinase 1 ; metabolism ; Mitogen-Activated Protein Kinase 3 ; metabolism ; Neutrophils ; cytology ; Reactive Oxygen Species ; metabolism ; Reperfusion Injury ; physiopathology ; Signal Transduction ; Umbilical Veins ; cytology

OBJECTIVETo study age and educational level and their relationship with fall-related injuries in Shanghai and to analyze the relevant costs.

METHODSMultistage cluster sampling was used for the selection of participants and standardized questionnaires were used for the information collection in 2006. Information on cases and deaths caused by fall-related injuries were obtained from 494 hospitals as well as from the mortality registry systems from 2001 till 2010.

RESULTSOf 45 857 participates, 674 suffered from fall-related injuries with the largest proportion among all injuries. The fall-related mortality increased from 10.63 per 100 000 in 2001 to 14.11 per 100 000 in 2010. The under-five mortality rate was the highest among children aged 0-14 years. Mortality increased dramatically among those aged 55 or above for the female and aged 60 or older for the male. Individuals with an educational level under the primary school were more likely to suffer fall-related injuries, accounting for 72.66% of all deaths and 49.24% of nonfatal cases respectively. The annual burden of fall-related injuries equated to 25.90% of the share of GDP for the healthcare, social security and welfare industries in 2006.

CONCLUSIONFall-related injuries were inversely related to victims' educational level. Children under the age of 5, women over 55 years old and men over 60 years old with an educational level lower than the primary school are the most risky groups of populations for intervention measures.

Accidental Falls ; mortality ; statistics & numerical data ; Adolescent ; Adult ; Age Distribution ; Aged ; Aged, 80 and over ; Child ; Child, Preschool ; China ; Educational Status ; Female ; Humans ; Male ; Middle Aged ; Sex Distribution ; Young Adult

OBJECTIVETo compare the effect and safety evaluation of catgut implantation at acupoint and drugs for levels of bone metabolism and free radicals in postmenopausal women.

METHODSSixty-five postmenopausal women were randomly divided into an acupoint catgut implantation group (33 cases) and a medication group (32 cases). Sanyinjiao (SP 6), Shenshu (BL 23) and Guanyuan (CV 4) were used as main points combined with adjunct points according to syndrome differentiation in the acupoint catgut implantation group, once two weeks. The medication group was treated with oral administration of 2 pills of Fu fuchun capsule, once a day. Three months constitute one course in the both groups. The scores of the symptoms and signs were evaluated, and elbow vein blood was drawn to detect the correlative index on the beginning and the ending day of the treatment respectively. The safety of catgut implantation at acupoint was evaluated after one course of treatment. Besides, the sex hormone test in ovulatory period was detected in twenty-eight normal women in reproductive age with regular menstrual cycle.

RESULTS1) The total effective rate of the acupoint catgut implantation group was 93.9% (31/33), and that of the medication group was 96.9% (31/32), there was no significant difference between the two groups (P>0.05). The contents of bone gla protein (BGP), calcitonin (CT), parathyroid hormone (PTH) and alkaline phosphatase (AKP) in the both groups had significant differences after treatment (P<0.05, P<0.01); 2) The content of estradiol (E2) in serum in the both groups was decreased more obviously than those of normal women in reproductive age (both P<0.001). The content of E2 in the both groups was obviously increased after treatment (P<0.05, P<0.01), this function in the acupoint catgut implantation group was weaker than that of the medication group (P<0.05), while the incidence rate of adverse effect in the acupoint catgut implantation group was lower than that of the medication group (P<0.05); 3) There were significant differences of the level of superoxide dismutase (SOD) and malondialdehyde (MDA) before and after treatment in the acupoint catgut implantation group, the level of SOD was obviously increased after treatment (P<0.01), while the level of MDA decreased obviously (P<0.001).

CONCLUSIONCatgut implantation at acupoint can improve the low level of estrogen of postmenopausal women, with good safety. It can regulate the levels of bone metabolism and free radicals of postmenopausal women, so it is very meaningful to prevent and treat postmenopausal degenerative diseases including the osteoporosis and to delay the process of apolexis.

Absorbable Implants ; adverse effects ; Acupuncture Points ; Adult ; Bone and Bones ; metabolism ; Catgut ; adverse effects ; Free Radicals ; metabolism ; Humans ; Middle Aged ; Postmenopause ; metabolism ; Treatment Outcome