Objective To construct the prokaryotic expression plasmid of HHV-8 fusion antigen for diagnosis of HHV-8 infec-tion .Methods The combined fragment ORF59 ,ORF65 and K8 .1 by fusion PCR was integrated into pQE-80L and transfected into E .coli DH5α.Fusion protein was induced to express by IPTG .SDS-PAGE and Western blot were employed to detect the fusion protein .Fusion protein was used to detect serum of blood donors .Results The combined plasmid pQE-80L-ORF59-ORF65-K8 .1 was constructed successfully after verifying by restriction enzyme digestion and sequencing .The fusion protein was about 24 KD and could be specific combined with HHV-8 positive serum .The fusion protein had the same result to detect HHV-8 with the HHV-8 ELISA kit .Conclusion Fusion protein we construct can be used as diagnosis antigen to detect HHV-8 of blood donors and common people .

ObjectiveTo observe the clinical effect and safety of direct-acting antiviral agents (DAAs) in patients with hepatitis C recurrence after liver transplantation. MethodsFifteen patients who received liver transplantation in 302 Hospital of PLA from December 2014 to May 2015 were selected, and after treatment, the HCV RNA-positive patients were treated with DAAs. The patients with genotype 1b were treated with Harvoni (sofosbuvir 400 mg/d + ledipasvir 90 mg/d) or sofosbuvir 400 mg/d + daclatasvir 60 mg/d, and those with genotype 2a were treated with sofosbuvir 400 mg/d + ribavirin 900mg/d. The course of treatment was 12 weeks. The changes in clinical symptoms and signs and laboratory markers including routine blood test, liver function, and HCV RNA quantification were observed regularly. ResultsAmong the 15 patients with hepatitis C recurrence after liver transplantation, 14 achieved HCV RNA clearance within 1-4 weeks, and the shortest time to clearance was 5 days; one patient had a HCV RNA level of 5.6×10 IU/ml at week 4. At month 12 of treatment, all the patients achieved HCV RNA clearance, and at the end of treatment, all the patients achieved virologic response. At present, 14 patients had achieved sustained virologic response for 12 weeks, and the other patients were still in the follow-up stage; at present, 3 patients had achieved sustained virologic response for 24 weeks, and the other 12 patients were still in the follow-up stage. Side effects of the treatment were mainly headache (1 case), weakness (5 cases), and arthralgia (1 case). ConclusionDAAs can be used in the treatment of patients with hepatitis C recurrence after liver transplantation with minor side effects, tolerance, and definite curative effect. Long-term sustained virologic response requires further observation.

OBJECTIVETo investigate the clinical manifestations, pathological characteristics, and treatments of urothelial-type mucinous adenocarcinoma of the prostate (UMAP).

METHODSWe reported a case of UMAP, reviewed relevant literature, and analyzed the clinicopaothological features, diagnosis, treatment, and prognosis of the disease.

RESULTSThe patient was a 60-year-old male and underwent transurethral resection of the prostate for dysuria. Postoperative pathology indicated mucinous adenocarcinoma and sigmoidoscopy revealed no primary colon cancer. Immunohistochemical staining showed the negative expressions of PSA and P504s and positive expressions of CK7, CK34 β E12, CK20, and CDX2. Thus UMAP was confirmed and treated by intensity-modulated radiotherapy. Then the patient was followed up for 30 months, which showed desirable therapeutic result, with neither local progression nor distant metastasis.

CONCLUSIONUMAP has a bad prognosis and its diagnosis depends on pathological and immunohistocchemical examinations. It responds well to radical prostatectomy but is not sensitive to endocrine therapy. Radiotherapy can be considered for those who are not fit to receive radical prostatectomy.

Adenocarcinoma, Mucinous ; metabolism ; pathology ; therapy ; Humans ; Keratins ; metabolism ; Male ; Middle Aged ; Neoplasm Proteins ; metabolism ; Prognosis ; Prostatectomy ; Prostatic Neoplasms ; metabolism ; pathology ; therapy ; Racemases and Epimerases ; metabolism

OBJECTIVETo investigate the relationship between the serum HBV DNA loads normalized to hepatic parenchyma cell volume and the liver histopathologic inflammation gradings in the immune clearance phase during the natural history of hepatitis B.

METHODSSerum HBV DNA loads were detected by fluorescence polymerase chain reaction and normalized to hepatic parenchyma cell volume. The association between normalized HBV DNA loads and liver inflammation histopathologic grade were analyzed.

RESULTSThe serum HBV DNA loads in patients with liver inflammation histopathologic grading 1, 2, 3 and 4 were 8.20*10(5)+/-9.11*10, 1.36*10(6)+/-5.96*10, 8.12*10(5)+/-8.01*10 and 2.08*10(6)+/-3.69*10 copies/ml, respectively (P more than 0.05). But the serum HBV DNA loads normalized to hepatic parenchyma cell volume in their located fibrosis stage were 9.24*10(8)+/-935, 5.33*10(9)+/-756, 1.06*10(10)+/-1770 and 3.31*10(11)+/-518 copies/ml, respectively (P less than 0.05).

CONCLUSIONThe serum HBV DNA load normalized to hepatic parenchyma cell volume in patients with different fibrosis stages is associated with liver histopathologic inflammation gradings.

Adult ; Automatic Data Processing ; Biopsy, Fine-Needle ; DNA, Viral ; blood ; Female ; Hepatitis B virus ; immunology ; isolation & purification ; Hepatitis B, Chronic ; blood ; immunology ; pathology ; virology ; Humans ; Inflammation ; pathology ; virology ; Liver Cirrhosis ; pathology ; virology ; Male ; Polymerase Chain Reaction ; methods ; Severity of Illness Index ; Viral Load ; Young Adult

OBJECTIVETo study the clinical characteristics of hepatitis B virus-related acute-on-chronic liver failure patients with familial aggregation.

METHODS275 patients with hepatitis B virus--related acute-on-chronic liver failure were investigated. The patients were divided into familial aggregation and non-familial aggregation group basis on their epidemiological features. Clinical data and biochemical indicators between the two groups were analyzed statistically.

RESULTS93 of 275 patients (33.82%) case were family aggregation. There was no significant difference compared with chronic hepatitis B patients (38.3%). The mean age of the two groups was 45.98 and 43.61 years old, respectively (P > 0.05). The rates of liver cirrhosis in family aggregation group were significant higher than non-familial aggregation group (73.91% vs 58.24%, p < 0.05). Serum total (TBil) and prothrombin activities (PTA) were no significant difference between the two groups, but ALT level in familial aggregation group was much higher (407.80 U/L vs 256.45 U/L, P 0.05).

CONCLUSIONFamilial aggregation were not related to acute-on-chronic liver failure in chronic HBV hepatitis patients. But the rate of liver cirrhosis were higher in patients with familial aggregation.

Acute Disease ; Adolescent ; Adult ; Aged ; Alanine Transaminase ; blood ; End Stage Liver Disease ; etiology ; genetics ; Family ; Female ; Hepatitis B ; complications ; Humans ; Liver Cirrhosis ; epidemiology ; Male ; Middle Aged

OBJECTIVETo investigate the expression of gene TRAIL driven by human telomerase reverse transcriptase (hTERT) promoter in SACC-83 cell tumor necrosis factor related apoptosis in dncing ligand.

METHODSAfter pACTERT-TRAIL plasmid transfected was into SACC-83 and HEL cells through liposome, the expression of TRAIL was examined using RT-PCR technique, the cells' survival rate by methyl thiazolyl tetrazolium (MTT) method and apoptosis rate by flow cytometry.

RESULTSExpression of extrinsic TRAIL gene driven by hTERT promoter was detected in SACC-83 cells, and not detected in human embryonic lung fibroblast (HEL) cells. After transfection of pACTERT-TRAIL, the proliferation of SACC-83 cells was significantly inhibited, and its apoptotic rate was promoted, whereas no inhibited effect was observed on HEL cells and its apoptotic rate showed little change.

CONCLUSIONShTERT promoter can be used to induce tumor-specific expression of TRAIL gene and apoptosis in SACC-83 cells.

Apoptosis ; Carcinoma, Adenoid Cystic ; genetics ; pathology ; Cell Line, Tumor ; Cell Proliferation ; Gene Targeting ; Genetic Vectors ; Humans ; Salivary Gland Neoplasms ; genetics ; pathology ; TNF-Related Apoptosis-Inducing Ligand ; genetics ; Telomerase ; genetics ; Transfection

OBJECTIVETo observe the effect of puerarin on cell proliferation, differentiation, maturation and mineralization in cultured rat osteoblasts.

METHODOsteoblasts from craniums of newly born SD rats were cultured in vitro. MTT, PNPP and ARS were used to observe the proliferation, activity of ALP and the number of mineral node of cultured osteoblasts in vitro.

RESULTIt was found that puerarin had the effect on stimulating cell proliferation, activity of ALP and the number of mineral node of cultured osteoblasts (P < 0.01 or P < 0.05).

CONCLUSIONPuerarin can promote proliferation, differentiation, maturation and mineralization of the osteoblasts in vitro.

Alkaline Phosphatase ; metabolism ; Animals ; Calcification, Physiologic ; drug effects ; Cell Differentiation ; drug effects ; Cell Proliferation ; drug effects ; Cells, Cultured ; Isoflavones ; pharmacology ; Osteoblasts ; cytology ; enzymology ; Rats ; Rats, Sprague-Dawley ; Skull ; cytology

Adseverin is a Ca2+-dependent actin filament-severing protein that has been reported to regulate exocytosis via rearrangements of the actin cytoskeleton in secretory cells. However, the role of adseverin in bone cells has not yet been well characterized. Here, we investigated the role of adseverin in osteoclastogenesis using primary osteoclast precursor cells. Adseverin expression was upregulated during RANKL (receptor activator of nuclear factor-kappaB ligand)-induced osteoclast differentiation. Moreover, genetic silencing of adseverin decreased the number of osteoclasts generated by RANKL. Adseverin knockdown also suppressed the RANKL-mediated induction of nuclear factor of activated T-cell c1 (NFATc1), which is a key transcription factor in osteoclastogenesis. In addition, adseverin knockdown impaired bone resorption and the secretion of bone-degrading enzymes from osteoclasts. These effects were accompanied by decreased NFATc1 expression and the activation of nuclear factor-kappaB. Collectively, our results indicate that adseverin has a crucial role in osteoclastogenesis by regulating NFATc1.
Active Transport, Cell Nucleus ; Animals ; Bone Resorption/genetics/metabolism/pathology ; Cell Differentiation ; Cells, Cultured ; Female ; Gelsolin/genetics/*metabolism ; Gene Knockdown Techniques ; Humans ; Mice, Inbred ICR ; NF-kappa B/metabolism ; NFATC Transcription Factors/*metabolism ; Osteoclasts/*cytology/metabolism/pathology ; RANK Ligand/*metabolism

This study aims to analyze the molecular epidemiological characteristics of HIV-1 strains prevailing among men who have sex with men (MSM) in Beijing, China. The pol gene fragments from 250 newly diagnosed HIV-1-infected MSM individuals during 2006-2010 in Beijing were amplified by RT-nested PCR, sequenced, and phylogenetically analyzed. HIV-1 pol gene from 189 individuals were amplified and analyzed; 81 (42. 9%), 3 (1. 6%), 2 (1.0%), 88 (46. 6%), and 15 (7.9%) individuals were infected with HIV-1 subtypes B, B', C, CRF01_AE, and CRF07_BC, respectively. The subtypes B and CRF01_AE could both be grouped into two clusters, and CRFO7_BC strains shared high homology and were presumed to originate from a common ancestor. The HIV-1 circulating in MSM in Beijing had a lower genetic diversity than in heterosexuals. The HIV-1 epidemic (2006-2010) in MSM in Beijing was actually a rapid spread of HIV-1 CRF01 AE and B, or rather native strains of the two viruses.
Adult ; China ; epidemiology ; Epidemics ; Genetic Variation ; HIV Infections ; diagnosis ; epidemiology ; virology ; HIV-1 ; classification ; genetics ; isolation & purification ; Homosexuality, Male ; statistics & numerical data ; Humans ; Male ; Middle Aged ; Molecular Epidemiology ; Molecular Sequence Data ; Phylogeny ; Young Adult

OBJECTIVETo explore the effect of Herba Epimedii flavone (HEF) on the osteoblast metabolism in vitro.

METHODOsteoblast were obtained from new born rat calvaria by digestive enzymes. MTF, PNPP and RT-PCR were used to observe the proliferation, activity of ALP and mRNA expression of OPG and RANKL of cultured osteoblasts in vitro.

RESULTIt was found that HEF had the effect on stimulating cell proliferation, activity of ALP and the mRNA expression of OPG of cultured osteoblasts (P < 0.01, P < 0.05).

CONCLUSIONHEF can promote the proliferation, the differentiation and the expression of OPG mRNA of the osteoblasts cultured in vitro.

Alkaline Phosphatase ; metabolism ; Animals ; Animals, Newborn ; Carrier Proteins ; biosynthesis ; genetics ; Cell Proliferation ; drug effects ; Cells, Cultured ; Epimedium ; chemistry ; Flavones ; isolation & purification ; pharmacology ; Glycoproteins ; biosynthesis ; genetics ; Membrane Glycoproteins ; biosynthesis ; genetics ; Osteoblasts ; cytology ; metabolism ; Osteoprotegerin ; Plants, Medicinal ; chemistry ; RANK Ligand ; RNA, Messenger ; biosynthesis ; genetics ; Rats ; Rats, Sprague-Dawley ; Receptors, Cytoplasmic and Nuclear ; biosynthesis ; genetics ; Receptors, Tumor Necrosis Factor ; biosynthesis ; genetics