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This study aimed to evaluate the efficacy and safety of entecavir(ETV) versus ETV maleate in Chinese patients with chronic hepatitis B(CHB). This was a randomized, double-blind, double-dummy, controlled, multicentre study. Patients were randomly assigned to receive 48 weeks of treatment with 0.5 mg/day ETV (group A) or 0.5 mg/day ETV maleate (group B), then, all patients received treatment with 0.5 mg/day ETV maleate from week 49 onwards. Patients were regularly followed up. Serum hepatitis B virus (HBV) markers were detected. Adverse events (AE) were recorded. The primary endpoint was the decline in HBV DNA in each group at the end of treatment. Secondary endpoints included the rate of HBV DNA below the lower limit of detection (LLOD) (20 I U/ml) at the end of treatment, the rate of hepatitis B e antigen (HBeAg) loss, the rate of HBeAg seroconversion and serum alanine aminotransferase (ALT) normalization. One hundred and thirty-seven (71 in group A) patients with HBeAg-positive CHB and 46 (21 in group A) patients with HBeAg-negative CHB completed the 240-week treatment and follow-up. Baseline characteristics were well balanced between the two groups. For the HBeAg-positive CHB patients, the mean HBV DNA level had similarly decreased from baseline in both groups (A: by 6.67 log

Background Currently, there are no drugs that have been proven to be effective against severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). Because of its broad antiviral activity, interferon (IFN) should be evaluated as a potential therapeutic agent for treatment of coronavirus disease 2019 (COVID-19), especially while COVID-19-specific therapies are still under development. Methods Confirmed COVID-19 patients hospitalized in the First Affiliated Hospital, School of Medicine, Zhejiang University in Hangzhou, China, from January 19 to February 19, 2020 were enrolled in a retrospective study. The patients were separated into an IFN group and a control group according to whether they received initial IFN-α2b inhalation treatment after admission. Propensity-score matching was used to balance the confounding factors. Results A total of 104 confirmed COVID-19 patients, 68 in the IFN group and 36 in the control group, were enrolled. Less hypertension (27.9% vs. 55.6%, P=0.006), dyspnea (8.8% vs. 25.0%, P=0.025), or diarrhea (4.4% vs. 19.4%, P=0.030) was observed in the IFN group. Lower levels of albumin and C-reactive protein and higher level of sodium were observed in the IFN group. Glucocorticoid dosage was lower in the IFN group (median, 40 vs. 80 mg/d, P=0.025). Compared to the control group, fewer patients in the IFN group were ventilated (13.2% vs. 33.3%, P=0.015) and admitted to intensive care unit (ICU) (16.2% vs. 44.4%, P=0.002). There were also fewer critical patients in the IFN group (7.4% vs. 25.0%, P=0.017) upon admission. Although complications during admission process were comparable between groups, the discharge rate (85.3% vs. 66.7%, P=0.027) was higher and the hospitalization time (16 vs. 21 d, P=0.015) was shorter in the IFN group. When other confounding factors were not considered, virus shedding time (10 vs. 13 d, P=0.014) was also shorter in the IFN group. However, when the influence of other factors was eliminated using propensity score matching, virus shedding time was not significantly shorter than that of the control group (12 vs. 15 d, P=0.206). Conclusions IFN-α2b spray inhalation did not shorten virus shedding time of SARS-CoV-2 in hospitalized patients.

Background A novel coronavirus called severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), first identified in Wuhan, China, has been rapidly spreading around the world. This study investigates the epidemiological and clinical characteristics of coronavirus disease 2019 (COVID-19) patients in Zhejiang Province who did or did not have a history of Wuhan exposure. Methods We collected data from medical records of confirmed COVID-19 patients in Zhejiang Province from Jan. 17 to Feb. 7, 2020 and analyzed epidemiological, clinical, and treatment data of those with and without recorded recent exposure in Wuhan. Results Patients in the control group were older than those in the exposure group ((48.19±16.13) years vs. (43.47±13.12) years, P

Introduction Elevated liver enzyme levels are observed in patients with coronavirus disease 2019 (COVID-19); however, these features have not been characterized. Methods Hospitalized patients with COVID-19 in Zhejiang Province, China, from January 17 to February 12, 2020, were enrolled. Liver enzyme level elevation was defined as alanine aminotransferase level >35 U/L for men and 25 U/L for women at admission. Patients with normal alanine aminotransferase levels were included in the control group. Reverse transcription polymerase chain reaction was used to confirm severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection, and patients symptomatic with SARS-CoV-2 infection were defined as patients with COVID-19. Epidemiological, demographic, clinical, laboratory, treatment, and outcome data were collected and compared. Results Of 788 patients with COVID-19, 222 (28.2%) patients had elevated liver enzyme levels (median [interquartile range {IQR}] age, 47.0 [35.0-55.0] years; 40.5% women). Being male, overweight, and smoking increased the risk of liver enzyme level elevation. The liver enzyme level elevation group had lesser pharyngalgia and more diarrhea than the control group. The median time from illness onset to admission was 3 days for liver enzyme level elevation groups (IQR, 2-6), whereas the median hospitalization time for 86 (38.7%) discharged patients was 13 days (IQR, 11-16). No differences in disease severity and clinical outcomes were noted between the groups. Discussion We found that 28.2% of patients with COVID-19 presented with elevated liver enzyme levels on admission, which could partially be related to SARS-CoV-2 infection. Male patients had a higher risk of liver enzyme level elevation. With early medical intervention, liver enzyme level elevation did not worsen the outcomes of patients with COVID-19.

As of Apr. 22, 2020, the World Health Organization (2020) has reported over 2.4 million confirmed coronavirus disease 2019 (COVID-19) patients and 169 151 deaths. Recent articles have uncovered genomic characteristics and clinical features of COVID-19 (Chan et al., 2020; Chang et al., 2020; Guan et al., 2020; Zhu et al., 2020), while our understanding of COVID-19 is still limited. As suggested by guidelines promoted by the General Office of National Health Commission of the People's Republic of China (2020) (from Versions 1 to 6), discharged standards for COVID-19 were still dependent on viral real-time polymerase chain reaction (RT-PCR) tests of respiratory specimens, showing that recovered COVID-19 patients with twice negative RT-PCR could meet discharge criteria. Here, we examined two cases in which nucleic acid test results were inconsistent with clinical and radiological findings, leading to suboptimal care.

Objective: The novel coronavirus pneumonia (COVID-19) has spread rapidly across the globe with the movement of people. How to diagnose COVID-19 quickly and accurately is a concern for all. We retrospectively assessed the clinical characteristics of patients with COVID-19 detected by outpatient screening in areas outside Wuhan, China, to guide early screening outside the epidemic area, to isolate and treat COVID-19-positive patients, and to control the spread of this virus in the region.Results: Among the 213 patients treated in the fever clinic of our hospital, 41 tested positive for novel coronavirus (2019-nCoV) and 172 were negative. Among the positive patients, 13 (31.7%) of the patients had been to Wuhan, while 28 (68.3%) had not been to Wuhan. There were 4 cases of clustering occurrence. The main symptoms exhibited by COVID-19-positive patients were fever (87.8%), cough (68.3%), and expectoration (34.1%). The C-reactive protein (CRP) levels were increased in 35 (85.3%) positive patients; the hydroxybutyrate dehydrogenase in the myocardial zymogram was increased in 22 positive patients (53.6%) and 38 negative patients (22.1%); computed tomography (CT) findings revealed lung lesions in all 41 positive patients (100%).Conclusion: We classified the patient population and analyzed the data to understand the early clinical performance of COVID-19. Our research illustrate that screening for COVID-19 outside Wuhan should focus on early symptoms such as fever and cough, in combination with lung CT findings, epidemiological history, and sputum pathogen detection to determine whether patients need further isolation.

A high fosfomycin resistance rate was observed in Klebsiella pneumoniae carbapenemase (KPC)-producing K. pneumoniae (KPC-KP) in our previous study, but little is known about its mechanisms. In this study, we explored the prevalence of plasmid-mediated fosfomycin resistance determinants among fosfomycin-resistant KPC-KP strains from a Chinese university hospital and determined the complete sequence of a novel fosA3 -carrying plasmid isolated from an epidemic K. pneumoniae sequence type (ST) 11 strain. A total of 97 KPC-KP strains were studied, of which 57 (58.8%) were resistant to fosfomycin, including 44 (45.4%) harboring fosA3 and 1 harboring fosA . All fosA3 -positive strains belonged to the dominant ST11-pulse type (PT) A clone according to multilocus sequence typing and pulsed-field gel electrophoresis, suggesting clonal dissemination. The fosA -positive isolate belonged to ST11-PTE. The fosA3 -carrying plasmid pKP1034 is 136,848 bp in length and is not self-transmissible. It is a multireplicon plasmid belonging to IncR-F33:A−: B−. Besides fosA3 , a variety of other resistance determinants, including bla KPC-2 , rmtB , bla CTX-M-65 , and bla SHV-12 , are identified in pKP1034, which would allow for coselection of fosA3 by most β-lactams and/or aminoglycosides and facilitate its dissemination despite limited use of fosfomycin in China. Detailed comparisons with related plasmids revealed that pKP1034 is highly mosaic and might have evolved from alarming recombination of the bla KPC-2 -carrying plasmid pKPC-LK30 from Taiwan and the epidemic fosA3 -carrying plasmid pHN7A8 from mainland China.

In the past few years, a growing body of clinical evidence has highlighted the risk of vitamin D deficiency in patients with chronic hepatitis C and that vitamin D levels are associated with the course of hepatitis C virus (HCV) infection, adverse effects, and treatment response to peginterferon/ribavirin. Recently, studies have found that vitamin D status is related to drug resistance and increased risk of infection in patients with liver cirrhosis. Vitamin D-related gene polymorphisms have been found to explain the interactions between vitamin D deficiency and HCV infection, offering a new perspective toward understanding the current problems such as the development of insulin resistance and racial differences in sustained virological response. Studies have been conducted to determine whether vitamin D supplementation as an adjuvant yields a better result compared with traditional HCV treatment. Here, we provide a brief review of the past and present knowledge of vitamin D in HCV infection.

Community-associated infections due to Escherichia coli producing CTX-M-type extended-spectrum β-lactamases are increasingly recognized in the United States. The bla CTX-M genes are frequently carried on IncF group plasmids. In this study, bla CTX-M-15 -harboring plasmids pCA14 (sequence type 131 [ST131]) and pCA28 (ST44) and bla CTX-M-14 -harboring plasmid pCA08 (ST131) were sequenced and characterized. The three plasmids were closely related to other IncFII plasmids from continents outside the United States in the conserved backbone region and multiresistance regions (MRRs). Each of the bla CTX-M-15 -carrying plasmids pCA14 and pCA28 belonged to F31:A4:B1 (FAB [FII, FIA, FIB] formula) and showed a high level of similarity (92% coverage of pCA14 and 99% to 100% nucleotide identity), suggesting a possible common origin. The bla CTX-M-14 -carrying plasmid pCA08 belonged to F2:A2:B20 and was highly similar to pKF3-140 from China (88% coverage of pCA08 and 99% to 100% nucleotide identity). All three plasmids carried multiple antimicrobial resistance genes and modules associated with virulence and biochemical pathways, which likely confer selective advantages for their host strains. The bla CTX-M -carrying IncFII-IA-IB plasmids implicated in community-associated infections in the United States shared key structural features with those identified from other continents, underscoring the global nature of this plasmid epidemic.

We sequenced a novel conjugative bla KPC-2 -harboring IncN plasmid, pYD626E, from an Escherichia coli sequence type 648 strain previously identified in Pittsburgh, Pennsylvania. pYD626E was 72,800 bp long and carried four β-lactamase genes, bla KPC-2 , bla SHV-12 , bla LAP-1 , and bla TEM-1 . In addition, it harbored qnrS1 (fluoroquinolone resistance) and dfrA14 (trimethoprim resistance). The plasmid profile and clinical history supported the in vivo transfer of this plasmid between Klebsiella pneumoniae and Escherichia coli .

Enterobacter amnigenus (EA76) and Klebsiella pneumoniae (KP76) isolates with multidrug-resistant (MDR) patterns were identified from the same patient in the neurosurgery department of our hospital. An outbreak of MDR K. pneumoniae had also occurred in this department. To characterize the resistance mechanism and molecular epidemiology of these isolates, sequential experiments including antimicrobial susceptibility testing, polymerase chain reaction (PCR), plasmid analysis, pulsed field gel electrophoresis (PFGE), and multilocus sequence typing (MLST) were performed. EA76 and KP76 were resistant to all of the antibiotics tested, except colistin and tigecycline. blaKPC-2, blaTEM-1, blaSHV-12, blaCTX-M-3, blaCTX-M-14, and rmtB genes were identified in both isolates, with blaKPC-2, blaTEM-1, blaCTX-M-14, and rmtB being co-carried on one plasmid in each isolate. Further analysis showed different restriction patterns between the two KPC-carrying plasmids. Of the 11 carbapenem-resistant isolates found in the outbreak, all were resistant to all of the β-lactams tested, with 63.64% (7/11) also exhibiting resistance to aminoglycosides and 72.73% (8/11) exhibiting resistance to quinolones. PCR analysis and molecular typing of the 11 K. pneumoniae strains revealed that the seven aminoglycoside-resistant isolates shared the same antibiotic-resistant gene pattern and identical or one-band-difference PFGE profiles relative to KP76. In addition, all of the eight aminoglycoside-resistant isolates, including KP76, belonged to the national epidemic clone ST11. The overall results indicate the emergence of E. amnigenus and outbreak of ST11 K. pneumoniae, with both co-harboring blaKPC and rmtB genes on a single plasmid in our neurosurgery wards.

The anti-allergic drug, N-(3,4-dimethoxycinnamonyl) anthranilic acid (3,4-DAA), is a synthetic anthranilic acid derivative that has been used therapeutically in Japan for many years. In this study, to investigate the effects of 3,4-DAA in allograft immunorejection model, liver orthotopic transplants were performed using inbred male Dark Agouti donors and Lewis rat recipients (allografts). The levels of indoleamine 2,3-dioxygenases (IDO) enzymic activities in five groups, allografts (control), dimethyl sulphoxide-treated group (vehicle control), 200 mg·kg–1·day–1 of 3,4-DAA-treated group and 200 mg·kg–1·day–1 of 3,4-DAA + 5 mg·ml–1 of 1-methyl-D-tryptophan (1-MT)-treated group were confirmed by determination of L-kynurenine (L-Kyn) concentrations. The serum alanine aminotransferase levels in 3,4-DAA-treated rats significantly decreased compared with those in mock and control group, whereas treatment of 1-MT in allografts led to the opposite effect. Administration of 3,4-DAA reduced histological severity of allograft immunorejection, decreased serum levels of cytokines tumour necrosis factor-alpha (TNF-α) and interferon-gamma (IFN-γ), and raised serum levels of interleukin-10 (IL-10), suggesting that 3,4-DAA has both anti-inflammatory and anti-immunorejection properties through IDO in immune regulation and may therefore be useful in filling an unmet need, in the treatment of allograft immunorejection. Copyright © 2011 John Wiley & Sons, Ltd.

Objective: To identify the correct site to biopsy in a case of pyrexia of unknown origin (PUO) caused by hepatic tuberculosis (TB). Clinical Presentation and Intervention: A 58-year-old man who developed hepatic TB presented with PUO. Ultrasonography (US) and computed tomography (CT) of the abdomen showed only calcifications in the liver, and positron emission tomography (PET)/CT showed diffuse increased metabolic activity in addition to focal areas of increased activity. A diagnosis of hepatic TB was confirmed by histological examination of liver tissues and interferon-γ release assays (IGRAs of T-SPOT/TB). The patient was treated with 4 anti-tubercular therapies (rifampicin, isoniazid, ethambutol and pyrazinamide). At the 3-month follow-up, the patient was disease free as confirmed by abdominal US. Conclusion: PET/CT was helpful in identifying a site to biopsy that led to the correct diagnosis.

Objective: To report a rare case of brucellosis with myelo-dysplastic syndrome (MDS). Clinical Presentation and Intervention: A 70-year-old woman presented with pancytopenia and fever of unknown origin (FUO). The initial diagnosis was brucellosis; the woman was treated with doxycycline and rifampin against Brucella melitensis but was later diagnosed as suffering from MDS. She was immediately transferred to the Department of Hematology for further evaluation. Conclusion: This study highlights the rarity of brucellosis with MDS, and we recommend that brucellosis with MDS be considered in patients presenting with pancytopenia and FUO.

To evaluate the efficacy and safety of entecavir maleate (ETV) versus ETV in Chinese patients with hepatitis B e antigen(HBeAg)-positive chronic hepatitis B(CHB).The patient population of this previously published randomized, double-blind, double-dummy, controlled, multicenter study was expanded by patients in the 0.5 mg/day ETV maleate group (total n = 110) and patients in the 0.5 mg/day ETV group (total n = 108). At treatment weeks 12, 24 and 48, hepatitis B virus (HBV) DNA levels were measured by the Roche Cobas Ampliprep/Cobas Taqman PCR assay. Adverse events (AE) were recorded.As in the original analysis, the two treatment groups showed similar characteristics at baseline. In addition, the results for the all therapeutic effects showed identical trends to the results obtained in the original analysis, including the statistically similar effects of ETV and ETV maleate treatment-induced decreases in mean HBV DNA level at weeks 12, 24, and 48 (ETV: by 4.28, 5.00, and 5.53 log10 IU/ml vs. ETV maleate: by 4.46, 4.99, and 5.51 log10 IU/ml, respectively; all vs. baseline P more than 0.05), achievement of undetectable levels of serum HBV DNA ( less than 20 IU/ml) at week 48 (ETV: 38.18% vs. ETV maleate: 35.19%; P more than 0.05), HBeAg loss rates at week 48 (ETV: 10.91% vs. ETV maleate: 12.96%; P more than 0.05), HBeAg seroconversion rates at week 48 (ETV: 7.77% vs. ETV maleate: 10.38%; P more than 0.05), normalization of alanine aminotransferase at week 48 (ETV: 75.47% vs. ETV maleate: 82.86%; P more than 0.05), and overall incidence of AE (ETV: 18.02% vs. ETV maleate: 17.43%; P more than 0.05).Performing analysis of the therapeutic efficacies of entecavir maleate versus entecavir with a larger study population confirmed our original findings of similar efficacy and safety profiles for these two drugs in patients with HBeAg-positive CHB.

Objective: To assess the efficacy of fluorine-18 fluorodeoxyglucose positron emission tomography (18F-FDG PET)/computed tomography (CT) in the diagnosis of patients with fever of unknown origin (FUO), who were finally diagnosed as lymphoma. Subjects and Methods: A retrospective study was performed in the First Affiliated Hospital, School of Medicine of Zhejiang University, China, from March 2009 to March 2012. The PET/CT images of consecutive patients with FUO were analyzed. Within 1 week of PET/CT scanning, additional histological tests were also performed if clinically needed. Results: A total of 73 consecutive patients were included. Of these, 34 (47%) had a PET/CT finding suggestive of the presence of lymphoma and 29 (85%) had a diagnosis of confirmed lymphoma; 39 (53%) had a PET/CT result revealing the absence of lymphoma and 4 (10%) were diagnosed by biopsy as having lymphoma, . The most frequent lymphoma diagnosis was peripheral T cell lymphoma (n = 16; 55%), followed by diffuse large B cell lymphoma (n = 9; 31%). The accuracy of PET/CT was 88%. Conclusion: In this study, PET/CT had high diagnostic accuracy in patients with FUO resulting from lymphoma, which indicated that PET/CT scanning was a valuable diagnostic tool for these groups of patients with FUO.

To investigate the therapeutic efficiency of antiviral treatment with pegylated-interferon (Peg-IFN) for hepatitis B e antigen (HBeAg)-positive chronic hepatitis B (CHB) and to explore whether liver histopathological features or other factors influence the HBeAg seroconversion treatment response.Eighty HBeAg-positive CHB patients with diagnosis confirmed by liver puncture were treated with Peg-IFN(2a or 2b)body weight dose, once weekly). At treatment week 48, the rate of HBeAg seroconversion was determined and used to analyze the influence of liver histopathological features (liver biopsy assessment of: inflammation, graded G0 to G4; fibrosis stage, graded S0 to S4), sex, age, differential levels (pre-treatment baseline vs. week 48 post-treatment) of serum alanine transferase (ALT), and HBV DNA, by binary logistic analysis.At week 48, the overall rate of HBeAg seroconversion was 30.0%. The rate of HBeAg seroconversion gradually advanced with increased liver inflammation (X2 = 8.435, P = 0.015): 9.09% of the 22 patients with G1; 31.58% of the 38 patients with G2; 47.30% of the 19 patients with G3; the one patient with G4. In contrast, the rate of HBeAg seroconversion showed a much weaker association with liver fibrosis (X2 = 5.917, P = 0.116). Only baseline HBeAg level, and no other baseline index, was significantly different between the patients who achieved HBeAg seroconversion and those who did not. Liver inflammation and baseline HBeAg level were identified as influencing factors of HbeAg seroconversion in response to Peg-IFN treatment.Peg-IFN therapy induces a higher rate of HBeAg seroconversion in HBeAg-positive CHB patients with severe liver inflammation; histological analysis of pre-treatment liver biopsies may help to identify patients most likely to benefit from the antiviral regimen.

Pentastomiasis is a type of parasitic zoonosis. Most patients with pentastomiasis are asymptomatic. We report here two pediatric patients with severe pentastomiasis (porocephaliais taiwan and armilliferiasis), and the results of their 6-year and 3-year follow-ups, respectively. The manifestations and outcomes of the two cases are described. The diagnoses were established by histopathologic and/or parasitologic examinations. After diagnosis, traditional Chinese medicine (TCM), as well as praziquantel and/or albendazole, were used for treatment. This report highlights the seriousness of pentastomiasis in children. We suggest TCM be considered as supplementary or even primary treatment of children with severe pentastomiasis.

The infection of Kaposi's sarcoma-associated herpes virus (KSHV) is most likely the cause of clinical Kaposi's sarcoma, primary effusion lymphoma, and multi-center Castleman's disease. KSHV infection has very limited epidemiological survey data in China, and its definite mode of transmission remains controversial. This study aimed to determine the infection status and the main transmission route of KSHV in Chinese population.An enzyme-linked immunosorbent assay (ELISA) utilizing KSHV ORF65 recombinant protein was employed to analyze the antibody response to KSHV ORF65 in sera from 122 healthy physical examination people, 107 intravenous drug users, 135 non-intravenous drug users, 211 hepatitis B (HBV) patients infected via blood transmission, 107 kidney transplant recipients, and 72 female sex workers in Zhejiang Province in Southeast China.KSHV infection occurred relatively common (13.1%) in healthy population in Zhejiang, China. Infection rate was 16.7% in female sex workers, but significantly elevated in intravenous drug addicts (58.9%), blood-transmitted HBV patients (28.0%) and kidney transplant patients (41.1%).Blood borne transmission of KSHV is probably the main route of infection in Zhejiang Province.

The anti-allergic drug, N-(3,4-dimethoxycinnamonyl) anthranilic acid (3,4-DAA), is a synthetic anthranilic acid derivative that has been used therapeutically in Japan for many years. In this study, to investigate the effects of 3,4-DAA in allograft immunorejection model, liver orthotopic transplants were performed using inbred male Dark Agouti donors and Lewis rat recipients (allografts). The levels of indoleamine 2,3-dioxygenases (IDO) enzymic activities in five groups, allografts (control), dimethyl sulphoxide-treated group (vehicle control), 200 mg·kg(-1) ·day(-1) of 3,4-DAA-treated group and 200 mg·kg(-1) ·day(-1) of 3,4-DAA + 5 mg·ml(-1) of 1-methyl-D-tryptophan (1-MT)-treated group were confirmed by determination of L-kynurenine (L-Kyn) concentrations. The serum alanine aminotransferase levels in 3,4-DAA-treated rats significantly decreased compared with those in mock and control group, whereas treatment of 1-MT in allografts led to the opposite effect. Administration of 3,4-DAA reduced histological severity of allograft immunorejection, decreased serum levels of cytokines tumour necrosis factor-alpha (TNF-α) and interferon-gamma (IFN-γ), and raised serum levels of interleukin-10 (IL-10), suggesting that 3,4-DAA has both anti-inflammatory and anti-immunorejection properties through IDO in immune regulation and may therefore be useful in filling an unmet need, in the treatment of allograft immunorejection.

Abnormal production of extracellular matrix (ECM) components significantly contributes to the development of liver fibrosis. This study aimed at examining the effects of short-hairpin RNA (shRNA)-mediated transient knockdown of transforming growth factor βRII (TGF βRII) expression on the proliferation and activation of hepatic stellate cells (HSCs) and synthesis of fibrogenic ECM components in HSC cells. Three different shRNA-expressing plasmids were constructed for the expression of shRNA-(a, b, c) targeting to the rat TGF βRII mRNA beginning at nucleotide position 339, 444 and 528 and they were transfected into a rat stellate cell line, HSC-T6 cells, respectively. The levels of TGF βRII, α-smooth muscle actin ( α-SMA), and type I and III collagen expressions were characterized by reverse transcription polymerase chain reaction and Western blot assays. The concentrations of hyaluronic acid (HA) and type IV collagen in the supernatants of cultured cells were measured by enzyme-linked immunosorbent assay. Transfection with the TGF βRII-specific shRNAs resulted in varying levels of inhibition in the expression of TGF βRII in HSC-T6 cells, and transfection with the potent shRNA-c inhibited the expression of TGF βRII in a dose-dependent manner. Knockdown of TGF βRII expression significantly reduced the levels of α-SMA, type I, III and IV collagen, and HA expression in HSC-T6 cells ( P < 0.01). In conclusion, our data indicated that knockdown of TGF βRII expression inhibited the activation of HSCs and the production of fibrogenic ECM components in HSC-T6 cells. Therefore, our findings support the notion that TGF βRII is an important factor of the pathogenic process of liver fibrosis.

Carbapenems are used to treat severe infections caused by multi-drug-resistant organisms, however, the emergence of carbapenem-resistant bacterial isolates is becoming an increasing therapeutic challenge. Since the first Klebsiella (K.) pneumoniae carbapenemase (KPC)-producing K. pneumoniae was reported in 2001, KPC-producing isolates have been found increasingly, specially in Enterobacteriaceae. The aim of this study was to characterize the mechanisms of a carbapenem-resistant Proteus (P.) mirabilis.A carbapenem-resistant P. mirabilis isolate was recovered from pleural drainage fluid of a patient admitted to surgical intensive care unit. Antimicrobial susceptibility testing of the isolate was performed by disk diffusion according to Clinical and Laboratory Standards Institute guidelines, and subsequent minimal inhibitory concentrations were determined with the E-test. Amplification of the bla(KPC) gene generated a positive band and the PCR products were sequenced subsequently. The plasmid of the isolate was extracted and was successfully transformed into Escherichia (E.) coli DH5α.The P. mirabilis isolate was resistant to all detected antimicrobial agents except tigecycline. KPC-2 was confirmed by DNA sequence analysis. The transformant E. coli was resistant to carbapenems. Further study demonstrated that upstream and downstream regions of bla(KPC-2) were identical to that observed in K. pneumoniae submitted to GenBank from China in 2007.Carbapenem resistance in the P. mirabilis isolate in this study is mainly due to production of KPC-2.

Augmenter of liver regeneration (ALR) is an important polypeptide in the process of liver regeneration. This study aimed to determine the expression level of ALR in different liver diseases and its significance.We prepared murine polyclonal antibody against ALR protein from Balb/C mice and purified the IgG fraction, which specifically combined to ALR protein as shown by Western blotting. Serum ALR levels in patients with hepatocellular carcinoma (HCC), hepatic failure (HF), chronic hepatitis B, and healthy persons were compared by ELISA. ALR mRNA expression levels in liver tissues in some of these patients were also compared by real-time RT-PCR. Immunohistochemical analysis was carried out on HF and HCC liver tissues.Different serum ALR levels foreshowed completely different prognoses in 18 HF patients. Higher ALR levels were noted in 6 improved patients (1613.5+/-369.6 pmol/ml) than in 12 deteriorating patients (462.3+/-235.8 pmol/ml). Similar levels were found in 20 HCC patients (917.9+/-332.7 pmol/ml), 24 chronic hepatitis B patients (969.2+/-332.5 pmol/ml) and 10 healthy persons (806.9+/-240.8 pmol/ml). ALR mRNA levels in HCC liver tissues [10E6.24 (1.74X10(6)) copies/μl] were much higher than in those of HF patients receiving orthotopic liver transplantation [10E3.45 (2.82X10(3)) copies/μl] or in healthy liver tissues [10E4.31 (2.04X10(4)) copies/μl]. In immunohistochemical analysis, positive immunostaining in HCC liver tissue was more intense than that in HF liver tissue.Serum ALR level is helpful in estimating the survival time of patients with HF, and ALR may play an important role in hepatocarcinogenesis.

To analyze the efficacy of adefovir dipivoxil combined with bicyclol in treatment of HBeAg-positive chronic viral hepatitis B (CHB).A total of 91 patients with CHB were randomized into experimental group and control group to be treated. The patients in experimental group (46 samples) received adefovir dipivoxil orally 10 mg daily and bicyclol orally 150 mg daily for 48 weeks and those in control group (45 samples) received adefovir dipivoxil orally 10 mg daily alone for 48 weeks. The serum aminotransferace (ALT/ AST), HBV-DNA, HBeAg/antiHBe were observed before and after treatment.Compared with pretreatment, the serum aminotransferace were all decreased obviously in two groups, the experimental group is better (P0.05). HBVDNA negative conversion rate was significantly higher in experimental group than in the control group (47.8% vs. 31.1%, P0.05). There were nostatistically differrence between the two groups in the portion of HBeAg loss rate and HBeAg seroconversion rate. There were no obvious adverse reaction in the study.Adefovir dipivoxil combined with bicyclol is efective in the treatment of chronic hepatitis B.

To investigate the relationship between clinical features of patients with A/H5N1 infection and their prognosis in mainland China.This study included 28 human cases with A/H5N1 infection in mainland China from October 2005 to May 2008. Data were collected and reviewed from hospital medical records and publishied papers. A database was built by EPIDATA 3.02 and statistical analyses were performed with SPSS 13.0.The median age of the 28 cases was 29 years (range 6-62), and 15 were females. Ten patients survived, and 18 died. The typically clinical manifestations of human influenza A/H5N1 infection included fever and lower respiratory infection. The numbers of peripheral white blood cells, lymphocytes and platelets in the survival and non-survival groups were (4.01 +/- 1.86) x 10(9)/L vs (5.1 +/- 2.9) x 10(9)/L, (1.09 +/- 0.49) x 10(9)/L vs (0.98 +/- 0.44) x 10(9)/L, and (116 +/- 39) x 10(9)/L vs (101 +/- 40) x 10(9)/L, respectively; the differences were not statistically significant between the 2 groups (P0.05). There was also no statistically significant difference in the increased serum enzymes, such as aspartate aminotransferase [(173 +/- 246) U/L vs (272 +/- 263) U/L], lactate dehydrogenase [(1016 +/- 568) U/L vs (1512 +/- 1052) U/L], creatine kinase [(1099 +/- 1590) U/L vs (2534 +/- 4281) U/L] and MB isoenzyme of creatine kinase [(28 +/- 30) U/L vs (125 +/- 197) U/L] (P0.05) between the survival and the non-survival groups. However, there was a statistically significant difference in the number of patients with an initial LDH level more than 8 fold of the normal value between the survival and the non-survival groups (none vs 6, P0.05). All of the 28 cases developed bilateral multiple infiltrates and consolidation in chest radiographs. Acute respiratory distress syndrome occurred in 22 cases, 17 of them died. All the 9 patients with acute kidney injury died. Ten patients received antiviral treatment with oseltamivir, and 6 of them survived. There was a statistical difference in the time of initiating oseltamivir treatment between the survival and the non-survival cases [(6.5 +/- 3.0) d vs (11.8 +/- 3.3) d, Z = 3.70, P0.05]. Broad spectrum antibiotics and corticosteroids were administered in all of the 28 cases. There was no statistical difference between the survival and the non-survival groups regarding to the corticosteroid treatment (P0.05).Initial LDH level reaching more than 8 fold of the normal value suggests a poor prognosis for human H5N1 infection. Patients complicated with either ARDS or acute kidney injury had a higher risk of death. Early administration of effective antiviral agents might improve the prognosis and decrease case fatality.

To investigate the expression and role of augmenter of liver regeneration (ALR) in hepatic failure.ALR polyclonal antibody was prepared and purified. Serum ALR in patients with hepatic failure, chronic hepatitis B and healthy persons were quantified by ELISA, ALR mRNA in hepatic tissues were quantified by real-time PCR.Different serum ALR levels foreshowed different outcomes for hepatic failure patients: The liver function was restored in 6 patients with higher ALR level [(1613.5+/-369.6) pmol/ml], and the liver function was deteriorated in 12 patients with lower ALR level [(462.3+/-235.8) pmol/ml]. ALR level in patients with chronic hepatitis B [(969.2+/-332.5) pmol/ml] was similar to that in healthy persons [(806.9+/-240.8) pmol/ml]. ALR mRNA level in hepatic failure patients receiving OLT (103.45 copies/microl) was lower than that in chronic hepatitis B patients (104.37 copies/microl) and healthy persons (104.31 copies/microl), ALR mRNA level in chronic hepatitis B and healthy persons was similar.These findings suggest serum ALR level reflected ALR mRNA level in liver and is helpful in estimating the survival time of patients with hepatic failure.

To investigate the intestinal microflora status and bacterial translocation in rats after liver transplantation.Male Brown-Norway (BN) rats were randomly divided into 4 groups: group I (n = 8) for liver transplantation; group II (n = 8) for simulated liver transplantation; group III (n = 8) for sham operation and group IV (n = 8) for normal group. Caecal bacterial counts, plasma endotoxin, intestinal mucosal ultrastructure and bacterial translocation to liver, spleen, kidney, and mesenteric lymph node were studied 24 h after surgery.The numbers of Bifidobacterium and Lactobacillus per gram of wet feces were significantly decreased in group I compare with those in the group III and group IV, while Enterobacteriaceae and Enterococcus counts were increased markedly compare with those in the group III and group IV, but no different was found between group I and group II. Impaired intestinal mucosa integrity were found in the group I and group II. In group I, the levels of plasma endotoxin increased after the transplantation when compare with group III and group IV. Increased incidence of bacterial translocation to liver, spleen and mesenteric lymph node were also observed after the transplantation (compare with those in the group IV, P0.01; compare with those in the group III, P0.01, P0.01, P0.05, separately). The increased rate of the bacterial translocation in liver was also found in transplantation group as compare with group II (P0.05).Liver transplantation may lead to disturbance of intestinal microflora and impairment of intestinal mucosal barrier function, and this dysfunction might be caused by the process of intestinal ischemia-reperfusion injury in transplantation.

To investigate the prevalence and associated risk factors of bacterial translocation (BT) in patients with cirrhosis after liver transplantation and analyze the effect of BT on bacterial infection after the surgery.Mesenteric lymph nodes (MLN), portal vein blood, and peripheral blood were collected during the liver transplantation for microbiological culture from 78 patients with cirrhosis. And meanwhile, all related clinical data were analyzed to investigate the risk factors of BT and its relationship with post-liver transplantation infections.BT was occurred in 8 of 78 cirrhotic patients (10.3%) and positive-rate of MLN culture was 5/8. Gram-negative aerobic bacillus was the main causative bacterium of BT (5/9), followed by Gram-positive aerobic enterococcus (22.2%, 2/9). Total bilirubin level in patients with BT was significantly higher than that in patients without BT.It suggests that hyperbilirubinemia is the only risk factor for BT, and BT is associated with an increased infectious rate after liver transplantation.

Hepatic ischemia/reperfusion injury may induce intestinal microflora imbalance. Salvia miltiorrhiza is effective in promoting blood circulation and counteracting peroxidation in tissues. The aim of the present study was to determine the effects of Salvia miltiorrhiza on intestinal microflora, endotoxemia, and bacterial translocation in rats with hepatic I/R injury.Sprague-Dawley rats in specific pathogen free grade were divided into 3 groups: group I(n=6) for sham operation; groups II(n=10) and III(n=7) for liver ischemia for 20 minutes and reperfusion for 22 hours. Group III was also pretreated with 4 ml/day of Salvia miltiorrhiza solution (250 mg/kg) by daily gavage for 7 days. The levels of serum alanine aminotransferase (ALT), aspartate aminotransferase (AST), malondialdehyde (MDA) and superoxide dismutase(SOD) in liver tissues, serum endotoxin, intestinal bacterial counts, intestinal mucosal histology and bacterial translocation were studied.The levels of ALT, AST, plasma endotoxin and MDA in liver tissues were decreased more markedly in group III (57.57+/-18.08 U/L, 147.57+/-40.84 U/L, 0.42+/-0.144 EU/ml and 0.52+/-0.19 nmol/mg-prot respectively) in group II(122.8+/-80.12 U/L, 295.9+/-216.92 U/L, 0.80+/-0.262 EU/ml and 0.72+/-0.12 nmol/mg-prot; P0.05-0.01 respectively). Liver SOD activity was increased more significantly in group III (318.47+/-64.62 U/mg-prot) than in group II(240.76+/-63.67 U/mg-prot, P0.05). The counts of Bifidobacteria and Bacteroides increased more significantly in group III than in group II, but were similar to those in group I. Bacterial translocation to the kidney in group II was 50%(5/10), whereas no bacterial translocation to the kidney occurred in the other two groups (P0.01). Ileal mucosal structure was markedly ameliorated in group III as compared with group II.Salviae miltiorrhiza could partially restore intestinal microflora balance, improve intestinal mucosal integrity, and reduce bacterial translocation and plasma endotoxin in rats with hepatic ischemia/reperfusion injury.

The demand for the clinical use of hepatocytes is increasing. The aim of this study was to develop a method for procurement of high qualitative pig hepatocytes and to evaluate the state of freshly isolated and cultured hepatocytes.The domestic extracorporeal circulating perfusion apparatus was used to isolate and harvest swine hepatocytes by the two-step perfusion method with EDTA and collagenase. The viability, function and morphology of the freshly isolated and cultured cells were evaluated and observed by the trypan blue exclusion test, biochemical measurements, phase contrast microscopy and transmission electron micrography (TEM).The total yield of isolated hepatocytes reached to 1.5(+/-0.4)X10(10) per liver with a viability of 92(+/-5)%, and the purity of hepatocytes reached to 98%. Immediately after isolation, phase-contrast microscope and TEM showed that undamaged hepatocytes appeared bright, translucent and spherical in shape, with a characteristic well-contrasted border. After 24 hours, the concentrations of alanine aminotransferase (ALT), aspartate aminotransferase (AST), lactate dehydrogenase (LDH), albumin (ALB), creatinine (Cr) and blood urea nitrogen (BUN) in the fluid of culture were declined significantly.This method of procuring swine hepatocytes could get high quality cells with active metabolic function.

AIM: To investigate the changes of gut microflora and endotoxin levels in rats with acute liver failure (ALF) induced by D-galactosamine (GalN). METHODS: Flora and endotoxin levels in the jejunum, ileum and colon in normal rats (group A) and rats with GalN-induced ALF were determined at 24 h (group B) or 48 h (group C) after GalN injection, as well as the endotoxin level in portal venous blood (PVB) and right ventricle blood (RVB) were determined by chromogenic limulus amoebocyte assay. RESULTS: Intestinal (jejunum, ileum, colon) lactobacillus count was statistically reduced in group B compared with those in group A (3.4 ± 0.3 vs 4.9 ± 0.3, 6.1 ± 0.4 vs 8.0 ± 0.3, 8.1 ± 0.2 vs 9.3 ± 0.2, P < 0.001, P < 0.001 and P < 0.001 respectively) and recovered partially in the group C compared with those in the group B, whereas the count of Enterobacteriaceae in the jejunum, ileum and colon in group B was increased markedly compared with those in the group A (5.1 ± 0.3 vs 3.6 ± 0.2, 6.9 ± 0.5 vs 5.3 ± 0.3, 8.7 ± 0.2 vs 7.6 ± 0.1, P < 0.001, P < 0.05 and P < 0.05 respectively) and restored partially in the group C compared with those in the group B. The endotoxin level in ileum was increased in the group B compared with those in the group A (111.3 ± 22.8 vs 51.5 ± 8.9, P < 0.05). In addition, the endotoxin level in PVB was obviously increased in group B compared with that in the group A (76.8 ± 9.1 vs 40.6 ± 7.3, P < 0.01) and reduced to the baseline at 48 h (group C). CONCLUSION: Severely disturbed gut flora in rats with GalN-induced acute liver failure plays an important role in the elevation of endotoxin level in PVB.