OBJECTIVETo explore the mechanism by which HBV X gene(HBx) inhibits apoptosis of human hepatoma cell line HepG2 in terms of miRNA.

METHODSThree cell lines were prepared: HepG2 cells stably transfected with HBx (HepG2/HBx), HepG2 cells stably transfected with pcDNA3.1 (HepG2/pcDNA3.1) and HepG2 cells. Flow cytometry was adopted to measure the apoptosis of these three cells and Taqman fluorescence quantitative PCR was used to examine miR-192 expression. After HepG2 cells was transfected with miR-192, the apoptosis was analyzed by flow cytometry and the expressions of p53 and PUMA at mRNA and protein levels were evaluated by SYBR Green quantitative PCR and Western blot, respectively.

RESULTSCompared with HepG2/pcDNA3.1 cells (11.46% ± 0.69%) and HepG2 cells (12.5% ± 0.66%), the apoptosis rate of HepG2/HBx cells (2.37% ± 0.35%) was significantly reduced (F = 171.722, P < 0.01). The level of miR-192 was 49.1% ± 5.9% in HepG2 cells, which was dramatically down-regulated (F = 14.319, P = 0.019) as compared to the other two groups (HepG2/pcDNA3.1: 98.0% ± 8.9%; HepG2: 100%). Compared with HepG2 cells transfected with miR-NC (10.74% ± 1.15%), transfection of miR-192 into HepG2 cells led to increased apoptosis (15.74% ± 1.17%) (F = 18.415, P = 0.013) and higher p53 and PUMA expressions at mRNA (p53: 1.68 ± 0.12 vs 0.90 ± 0.09, F = 43.115, P = 0.003, PUMA: 1.66 ± 0.10 vs 0.98 ± 0.06, F = 22.541, P = 0.009) and protein (p53: 3.07 vs 1, PUMA: 2.13 vs 1) levels.

CONCLUSIONHBx could inhibit apoptosis of HepG2 cells through down-regulation of miR-192 which induces apoptosis of HepG2 cells.

Apoptosis ; Carcinoma, Hepatocellular ; genetics ; metabolism ; pathology ; Down-Regulation ; Genes, Viral ; Hep G2 Cells ; Hepatitis B virus ; genetics ; Humans ; Liver Neoplasms ; genetics ; metabolism ; pathology ; MicroRNAs ; metabolism ; Trans-Activators ; genetics ; metabolism

Objective:To study correlation between carotid atherosclerosis(AS)stenotic severity and myocardial infarction(MI)size assessed by carotid color ultrasonography.Methods:According to infarction size,a total of 180 MI patients trea-ted in our hospital were divided into focal necrosis group(n=92),small-medium size infarction group(n=45)and large size infarction group(n=43).Carotid artery stenosis,peak systolic flow velocity(PSV),end diastolic flow velocity(EDV),number of AS plaques and MI location were compared among all group.And correlation between carotid AS ste-notic severity and MI size was analyzed.Results:Compared with focal necrosis group,there were significant reductions in PSV[(101.43±23.16)cm/s vs.(154.35±65.73)cm/s vs.(325.19±105.27)cm/s]and EDV[(34.19±19.36)cm/s vs.(77.13±20.61)cm/s vs.(138.41±84.93)cm/s]in small-medium size infarction group and large size infarction group,and above indexes of large size infarction group were significantly higher than those of small-medium size infarction group(P=0.001 all).Compared with focal necrosis group and small-medium size infarction group,there were significant rise in percentages of severe carotid stenosis(4.35%,11.11%vs.37.27%),occlusion(1.09%,4.44%vs.27.91%),number of atherosclerotic plaque＞2(38.04%,40.00%vs.65.12%)and anterior infarction(32.61%,35.56%vs.69.77%),and significant reduction in percentage of posterior infarction(48.91%,44.44%vs.23.26%)in large size infarction group(P＜0.05 or＜0.01).Spearman correlation analysis indicated that carotid AS stenotic severity was significant positively cor-related with MI size(r=0.733,P=0.001).Conclusion:Incidence rates of severe carotid stenosis and occlusion are higher in patients with large size myocardial infarction.Carotid AS stenotic severity is significant positively correlated with myo-cardial infarction size,which possesses certain application value in detecting myocardial infarction.

Background: As a traditional Chinese medicine, Cordyceps sinensis (CS) possesses a variety of immunoregulatory properties. This study aimed to explore the therapeutic potential of CS in a mice model of multiple sclerosis (MS)?experimental autoimmune encephalomyelitis (EAE). Methods: Female C57BL/6 mice were immunized with myelin oligodendrocyte glycoprotein35–55 to induce EAE, followed by an instant intragastric feeding with a low dosage of CS (low?CS group, n = 5), high dosage of CS (high?CS group, n = 5), or the same volume of normal saline (control group, n = 5).All the mice were observed for clinical assessment. Over the 30 days of CS treatment, flow cytometry was used to detect the frequency of helper T?cell (Th) subsets, Th1 and Th17, and CD4+ CD25+ regulatory T cells in the spleen and lymph nodes. Meanwhile, pathological changes in brain were determined using both hematoxylin?eosin and luxol fast blue staining. Data were analyzed using the one?way analysis of variance (ANOVA). Results: Over the 15 and 30 days of CS treatment, the clinical assessment for EAE demonstrated that both high?CS group (2.51 ± 0.31 and 2.26 ± 0.39 scores, respectively) and low?CS group (2.99 ± 0.40 and 2.69 ± 0.46, respectively) had lower disease severity scores than those of control group (3.57 ± 0.53 and 3.29 ± 0.53, all P < 0.01, respectively). Meanwhile, after 15 and 30 days, the high?CS group (19.18 ± 1.34 g and 20.41 ± 1.56 g, respectively) and low?CS group (18.07 ± 1.18 g and 19.48 ± 1.69 g, respectively) had a lower body weight, as compared with control group (16.85 ± 1.15 g and 18.22 ± 1.63 g, all P < 0.01, respectively).At 30 days post?CS treatment, there was a lower Th1 frequency in the lymph nodes (2.85 ± 1.54% and 2.77 ± 1.07% vs. 5.35 ± 1.34%, respectively; P < 0.05) and spleens (3.96 ± 1.09% and 3.09 ± 0.84% vs. 5.07 ± 1.50%, respectively; P < 0.05) and less inflammatory infiltration and demyelination in the brain of CS?treated mice than that of control group. Conclusions: Our preliminary study demonstrated that CS efficiently alleviated EAE severity and EAE?related pathology damage and decreased the number of Th1s in the periphery, indicating its effectiveness in the treatment of murine EAE. Thus, our findings strongly support the therapeutic potential of this agent as a new traditional Chinese medicine approach in MS treatment.

OBJECTIVETo provide experimental data for the quality control of processed Paeonia lactiflora, a Chinese herbal medicine.

METHODTraditional processing of P. lactiflora was simulated, content of paeoniflorin and water extracts among different preparations were assayed by HPLC; The quantitative correlations among different processing conditions were analyzed, the effects of processing parameters on the contents of paeoniflorin and water extracts were assayed a nanalysed.

RESULT AND CONCLUSIONThe controlled processing parameters were correlated with covariables which showed that processing procedures was controllable, and the heating temperature was a factor impacting the content of paeoniflorin.

Benzoates ; analysis ; Bridged-Ring Compounds ; analysis ; Glucosides ; analysis ; Hot Temperature ; Monoterpenes ; Paeonia ; chemistry ; Plants, Medicinal ; chemistry ; Quality Control ; Technology, Pharmaceutical ; methods ; Temperature

Adult ; Cerebral Infarction ; etiology ; prevention & control ; Female ; Humans ; Immunosuppressive Agents ; adverse effects ; Intracranial Hemorrhages ; etiology ; prevention & control ; Liver Transplantation ; adverse effects ; Male ; Middle Aged ; Tacrolimus ; adverse effects

OBJECTIVETo investigate the rule of lymph node metastasis of adenosquamous carcinoma of the lung.

METHODSThe data of 361 surgically treated patients with adenosquamous carcinoma of the lung from October 1965 to June 2003 were collected and retrospectively reviewed. The classification of regional lymph node stations and TNM stage were determined according to the UICC criteria (1997). The route and patterns as well as influencing factors of lymph node metastasis were analyzed by SPSS 10.0 software. The median follow-up period was 5.5 years (range, 1.4 to 23.4 years).

RESULTSThe analysis of the route of mediastinal lymph node metastasis in the 361 cases showed that the tumor originated in the left upper lobe firstly metastasized to station 5 (A-P window), tumor in the right upper lobe to the station 4 (lower paratracheal), then secondly to station 7 (subcarinal), lastly to station 3 from the tumor in the left upper lobe or to the station 2 from the tumor in the right upper lobe. It was found that the tumors originated from the lower lobe, firstly metastasized to station 7, secondly to station 9 or 4 from the right lobe; or station 5 from left lower lobe, lastly to station 3 or 2 in the mediastinum. For the tumor in the middle lobe, mainly metastasized to station 7, 4 and 2. The skip mediastinal lymph node metastasis but N1 negative most commonly metastasized to station 7, then to station 4 from the tumor in the right lung and 5 from the tumor in the left lung. The prognosis of patients with a single skipping metastasis to mediastinal lymph node (N1-, SMLN) was better than that in the other patients with mediastinal lymph node metastases.

CONCLUSIONThe lung cancer growing in a different location has a different route and skipping metastasis to mediastinal lymph nodes. The patterns of lymph node metastasis affect prognosis. The prognosis of patients with single skipping metastasis to mediastinal lymph nodes but negative pulmonary hilar lymph node is better than that in the other patients with multiple station mediastinal lymph node metastases. The "N1-, SMLN" pattern ought to be considered as a special lymph nodal metastasis with better prognosis.

Adult ; Aged ; Aged, 80 and over ; Carcinoma, Adenosquamous ; pathology ; surgery ; Female ; Follow-Up Studies ; Humans ; Lung Neoplasms ; pathology ; surgery ; Lymph Node Excision ; Lymph Nodes ; pathology ; surgery ; Lymphatic Metastasis ; Male ; Mediastinum ; Middle Aged ; Neoplasm Staging ; Pneumonectomy ; Retrospective Studies ; Survival Rate

To investigate the effect of hepatitis B virus X protein(HBx) on CtBP-interacting protein(CtIP) which is an important repair factor of DNA double strand break damage in HepG2 cells induced by bleomycin. A HBx stably expressing HepG2 cell line and a control HepG2 cell line with empty vector transfected were established. After the double strand break (DSB) damage occurred, the mRNA and protein levels of CtIP were detected by Real-time PCR and Western blot assay respectively, cell cycle profiles and apoptotic cell death were determined by a flow cytometry, and the position of CtIP in cells was observed by confocal laser scanning microscopy. It showed that HepG2 cells transfected with hepatitis B virus X gene could stably express HBx protein. After being induced by bleomycin, the percentage of apoptotic cell was 16.90%+/-0.89% in HBx stably expressing HepG2 cell line and 15.30%+/-0.86% in control cell line, respectively (q = 2.074, P is more than to 0.05). While the percentage of death cell was 8.71%+/-0.74% in HBx stably expressing HepG2 cell line and 4.90%+/-0.46% in control cell line, respectively (q = 7.126, P is less than to 0.01). The two cell lines manifested the increase of G2/M arrest and significant difference existed between the two cell lines. HBx down regulated the expression levels of CtIP and its mRNA. The CtIP level was 0.66+/-0.04 in HepG2-HBx cell and 0.73+/-0.05 in HepG2-vec cell, respectively (t = 2.314, P is less than to 0.05). The relative mRNA level was 1.00+/-0.06 in HepG2-HBx cell and 1.23+/-0.08 in HepG2-vec cell, respectively (t = 2. 732, P is less than to 0.05). We also found that CtIP was concentrated in the cell nucleus. The research suggests that HBx may affect DNA-repair pathways by disrupting the expression of CtIP.
Carcinoma, Hepatocellular ; metabolism ; Hep G2 Cells ; Hepatitis B virus ; genetics ; Humans ; Liver Neoplasms ; metabolism ; Real-Time Polymerase Chain Reaction

OBJECTIVETo explore the effect of one dressing of porcine acellular dermal matrix on deep partial thickness burns.

METHODSFrom January 1997 to January 2004, sixty-seven cases of deep partial thickness total burned surface area (TBSA) from 50% to 90% burn wound were treated by a single dressing of porcine acellular dermal matrix (the porcine acellular dermal matrix group). Ten cases of deep partial thickness burned patients with the same TBSA treated by exposure method served as the exposure method group. The healing time of the wound was observed. The patients were followed up for 3 months to 2 years, and the scar proliferation was observed.

RESULTSThe deep partial-thickness wound would be healed without dressing change in the porcine acellular dermal matrix group, and the average healing time was (12.2 +/- 2.6) days. The average healing time of the exposure method group was (27.4 +/- 3.5) days. Follow up of the patients within 3 months to 2 years showed that scar proliferation in the porcine acellular dermal matrix group was much less than that in the exposure method group, even no scar proliferation was observed in some patients.

CONCLUSIONWithout tangential excision, autografting and dressing change, a single dressing of porcine acellular dermal matrix on deep partial thickness burn wound could shorten the healing time and inhibit scar proliferation.

Animals ; Biological Dressings ; Burns ; pathology ; therapy ; Cicatrix ; prevention & control ; Female ; Follow-Up Studies ; Humans ; Male ; Swine ; Treatment Outcome ; Wound Healing

OBJECTIVETo investigate the feasibility and effectivity of using compound material of fibrin glue and DBM as scaffolds for cartilage tissue engineering.

METHODSChondrocytes isolated from articular cartilage were seeded into prepared scaffolds, after incubation for 4 weeks in vitro. Chondrocytes and fibrin glue and DBM constructs were implanted in the joint cave of rabbit. The specimens were excised at the 4th, 8th, 12th week, examined grossly analyzed by haematoxylin cosine, toluidine blues staining and type II collagen immunohistochemistry reaction. Wakitani score was counted to evaluate the repairing effect.

RESULTSGrossly analysis showed some ivory tissue filled the caves after 4 weeks and the caves were full filled with smooth surface after 12 weeks. The microscope showed a good deal of chondrocytes appeared after 8 weeks and more type II collagen than 4 weeks. Twelve weeks later, cartilage lacuna could be observed. The cells arrangement and the amount of type II collagen both showed the same as the natural one.

CONCLUSIONComplicated material of fibrin glue and DBM as scaffolds can be used as scaffolds for cartilage tissue engineering.

Animals ; Bone Matrix ; physiology ; transplantation ; Cartilage, Articular ; physiopathology ; surgery ; Cell Transplantation ; Cells, Cultured ; Chondrocytes ; physiology ; transplantation ; Female ; Fibrin Tissue Adhesive ; therapeutic use ; Male ; Rabbits ; Random Allocation ; Regeneration ; Tissue Adhesives ; therapeutic use ; Tissue Engineering ; Tissue Scaffolds

OBJECTIVETo study the HIV incidence and risk factors among sero-negative spouses of HIV patients in Dehong prefecture of Yunnan province.

METHODSA cohort of sero-negative spouses of the HIV patients had been developed and followed up since November, 2005. HIV new infections and related behaviors had been investigated every six months.

RESULTSBy the end of June, 2008, 790 sero-negative spouses of HIV patients had been recruited, of whom 702 were followed-up for at least one time. During the total 1202.35 person-years, 31 new HIV infections were identified, with an overall incidence of 2.58/100 person-years. The HIV incidence rates were 2.22/100 person-years in 2006, 2.95/100 person-years in 2007 and 2.74/100 person-years in 2008. Data from the Cox proportional hazard regression model indicated that those who resided in Yingjiang county [hazard ratio (HR) = 4.37, 95%CI: 1.48 - 12.90, P = 0.008], ever using drugs (HR = 3.49, 95%CI: 1.09 - 11.18, P = 0.035), or having an HIV-infected spouse who never exposed to antiretroviral treatment (HR = 3.60, 95%CI: 1.41 - 9.16, P = 0.007) were at higher risk for HIV infection.

CONCLUSIONSero-negative spouses of HIV patients in Dehong prefecture of Yunnan province had a relatively high incidence of HIV new infection during 2006-2008. More efforts should put on those people living in these areas, having a history of drug use or having an HIV-infected spouse who had never been exposed to antiretroviral treatment.

Adolescent ; Adult ; China ; epidemiology ; Female ; HIV Infections ; epidemiology ; HIV Seronegativity ; Humans ; Incidence ; Male ; Middle Aged ; Prospective Studies ; Risk Factors ; Sexual Behavior ; Spouses ; Young Adult