Safety of protease inhibitors and Arbidol for SARS-CoV-2 pneumonia in Zhejiang Province, China / 浙江大学学报（英文版）（B辑：生物医学和生物技术）

The aim of this study was to evaluate the safety of an antiviral regimen of protease inhibitors combined with Arbidol (umifenovir) for severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) pneumonia patients. The genomic sequence of SARS-CoV-2 is highly homologous to that of SARS-CoV (Zhou et al., 2020). Previously published basic and clinical research on anti-SARS-CoV treatment found that lopinavir/ritonavir (LPV/r) could improve the prognosis of SARS patients (Chan et al., 2003; Chu et al., 2004). Darunavir (DRV) is another protease inhibitor that blocks the binding of SARS-CoV-2 to human angiotensin-converting enzyme 2 (Omotuyi et al., 2020). The broad-spectrum antiviral drug Arbidol (umifenovir) also shows in vitro anti-SARS-CoV activity (Khamitov et al., 2008).

Comparison of epidemiological and clinical characteristics of COVID-19 patients with and without Wuhan exposure history in Zhejiang Province, China / 浙江大学学报（英文版）（B辑：生物医学和生物技术）

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<0.001), and more were over 65 years old (15.95% control vs. 5.60% exposure, P<0.001). The rate of clustered onset was also significantly higher in the control group than in the exposure group (31.39% vs. 18.66%, P<0.001). The symptom of a sore throat in patients in the exposure group was significantly higher than that in the control group (17.30% vs. 10.89%, P=0.01); however, headache in the exposure group was significantly lower than that in the control group (6.87% vs. 12.15%, P=0.015). More patients in the exposure group had a significantly lower level of lactate dehydrogenase (LDH) and aspartate aminotransferase (AST) than those in the control group. There was no significant difference in any degree of COVID-19 including mild, severe, and critical between the two groups.@*CONCLUSIONS@#From the perspective of epidemiological and clinical characteristics, there was no significant difference between COVID-19 patients with and without Wuhan exposure history.

Transmission risk of patients with COVID-19 meeting discharge criteria should be interpreted with caution / 浙江大学学报（英文版）（B辑：生物医学和生物技术）

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.

Interferon-α2b spray inhalation did not shorten virus shedding time of SARS-CoV-2 in hospitalized patients: a preliminary matched case-control study / 浙江大学学报（英文版）（B辑：生物医学和生物技术）

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.

Utility of fluorodeoxyglucose positron emission tomography/computed tomography in patients with fever of unknown origin diagnosed as lymphoma

To assess the efficacy of fluorine-18 fluorodeoxyglucose positron emission tomography [[18]F-FDG PET]/computed tomography [CT] in the diagnosis of patients with fever of unknown origin [FUO], who were finally diagnosed as lymphoma. 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. 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%. 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

Liver Tuberculosis presenting as an uncommon cause of pyrexia of unknown origin: positron emission tomography/computed tomography identifies the correct site for biopsy

To identify the correct site to biopsy in a case of pyrexia of unknown origin [PUO] caused by hepatic tuberculosis [TB]. 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-Y 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. PET/CT was helpful in identifying a site to biopsy that led to the correct diagnosis

Analysis of a randomized, double-blind, double-dummy, controlled, multicenter study confirmed the similar therapeutic efficacies of entecavir maleate and entecavir for treatment of HBeAg-positive chronic hepatitis B / 中华肝脏病杂志

<p><b>OBJECTIVE</b>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).</p><p><b>METHODS</b>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.</p><p><b>RESULTS</b>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).</p><p><b>CONCLUSION</b>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.</p>

Liver histopathological features influencing HBeAg seroconversion in patients with HBeAg-positive chronic hepatitis B responding to Peg-IFN treatment / 中华肝脏病杂志

<p><b>OBJECTIVE</b>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.</p><p><b>METHODS</b>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.</p><p><b>RESULTS</b>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.</p><p><b>CONCLUSION</b>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.</p>

Acute Brucellosis with Myelodysplastic syndrome presenting as pancytopenia and fever of unknown origin

To report a rare case of brucellosis with myelo-dysplastic syndrome [MDS]. 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. 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

Regional prevalence and transmission route of Kaposi's sarcoma-associated herpes virus in Zhejiang, China / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>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.</p><p><b>METHODS</b>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.</p><p><b>RESULTS</b>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%).</p><p><b>CONCLUSION</b>Blood borne transmission of KSHV is probably the main route of infection in Zhejiang Province.</p>

Emergence of Klebsiella pneumoniae carbapenemase-producing Proteus mirabilis in Hangzhou, China / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>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.</p><p><b>METHODS</b>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.</p><p><b>RESULTS</b>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.</p><p><b>CONCLUSION</b>Carbapenem resistance in the P. mirabilis isolate in this study is mainly due to production of KPC-2.</p>

The analyse of effectiveness in HBeAg-positive chronic viral hepatitis B treated by adefovir dipivoxil combined with bicyclol / 中华实验和临床病毒学杂志

<p><b>OBJECTIVE</b>To analyze the efficacy of adefovir dipivoxil combined with bicyclol in treatment of HBeAg-positive chronic viral hepatitis B (CHB).</p><p><b>METHODS</b>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.</p><p><b>RESULTS</b>Compared with pretreatment, the serum aminotransferace were all decreased obviously in two groups, the experimental group is better (P < 0.05). HBVDNA negative conversion rate was significantly higher in experimental group than in the control group (47.8% vs. 31.1%, P < 0.05). 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.</p><p><b>CONCLUSION</b>Adefovir dipivoxil combined with bicyclol is efective in the treatment of chronic hepatitis B.</p>

Expression and role of augmenter of liver regeneration in patients with hepatic failure / 中华肝脏病杂志

<p><b>OBJECTIVE</b>To investigate the expression and role of augmenter of liver regeneration (ALR) in hepatic failure.</p><p><b>METHODS</b>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.</p><p><b>RESULTS</b>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.</p><p><b>CONCLUSION</b>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.</p>

The change of intestinal microecology in rats after orthotopic liver transplantation / 中华外科杂志

<p><b>OBJECTIVE</b>To investigate the intestinal microflora status and bacterial translocation in rats after liver transplantation.</p><p><b>METHODS</b>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.</p><p><b>RESULTS</b>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, P < 0.01; compare with those in the group III, P < 0.01, P < 0.01, P < 0.05, separately). The increased rate of the bacterial translocation in liver was also found in transplantation group as compare with group II (P < 0.05).</p><p><b>CONCLUSIONS</b>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.</p>

Investigation of intestinal bacterial translocation in 78 patients with cirrhosis after liver transplantation / 中华外科杂志

<p><b>OBJECTIVE</b>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.</p><p><b>METHODS</b>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.</p><p><b>RESULTS</b>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.</p><p><b>CONCLUSIONS</b>It suggests that hyperbilirubinemia is the only risk factor for BT, and BT is associated with an increased infectious rate after liver transplantation.</p>

Lipopolysaccharide (LPS) increases tumor necrosis factor-alpha related apoptosis induced-ligand (TRAIL) in macrophages killing HepG2 cells / 中华肝脏病杂志

<p><b>OBJECTIVE</b>To investigate the influence of lipopolysaccharide (LPS) on macrophages expressing TNF-alpha related apoptosis induced-ligand (TRAIL) and its relation to apoptosis of HepG2 cell line.</p><p><b>METHODS</b>Membrane-bound TRAIL (mTRAIL) was measured by flow cytometry; soluble TRAIL in supernatant was detected by enzyme-linked immunoabsorbent sandwich assay (ELISA); cytotoxicity of TRAIL to HepG2 cell line was measured by chromium release assay, and apoptosis of HepG2 cell was confirmed by Annexin V staining.</p><p><b>RESULTS</b>LPS only slightly increased membrane-bound TRAIL expression of macrophages. On the other hand, soluble TRAIL in the supernatant was increased with LPS stimulation, and the optimal concentration of LPS was 100 ng/ml (sTRAIL value 67.40 ng/ml+/-5.08 ng/ml). The soluble TRAIL in the supernatant was cytotoxic to HepG2 cells, and this activity can be blocked by TRAIL neutralizing antibodies.</p><p><b>CONCLUSION</b>LPS increases the expression of soluble TRAIL in macrophages, and soluble TRAIL is toxic to HepG2 cells. All of our results indicate that TRAIL may play an important role in the pathogenesis of viral hepatitis.</p>

Intestinal microflora in rats with ischemia/reperfusion liver injury / 浙江大学学报（英文版）（B辑：生物医学和生物技术）

<p><b>OBJECTIVES</b>To investigate the intestinal microflora status related to ischemia/reperfusion (I/R) liver injury and explore the possible mechanism.</p><p><b>METHODS</b>Specific pathogen free grade Sprague-Dawley rats were randomized into three groups: Control group (n=8), sham group (n=6) and I/R group (n=10). Rats in the control group did not receive any treatment, rats in the I/R group were subjected to 20 min of liver ischemia, and rats in the sham group were only subjected to sham operation. Twenty-two hours later, the rats were sacrificed and liver enzymes and malondialdehyde (MDA), superoxide dismutase (SOD), serum endotoxin, intestinal bacterial count, intestinal mucosal histology, bacterial translocation to mesenteric lymph nodes, liver, spleen, and kidney were studied.</p><p><b>RESULTS</b>Ischemia/reperfusion increased liver enzymes, MDA, decreased SOD, and was associated with plasma endotoxin elevation in I/R group compared to those in the sham group. Intestinal Bifidobacterium and Lactobacillus decreased and intestinal Enterobacteria and Enterococci, bacterial translocation to kidney increased in the I/R group compared to the sham group. Intestinal microvilli were lost, disrupted and the interspace between cells became wider in the I/R group.</p><p><b>CONCLUSION</b>I/R liver injury may lead to disturbance of intestinal microflora and impairment of intestinal mucosal barrier function, which contributes to endotoxemia and bacterial translocation to kidney.</p>

Phenotypic and functional characteristics of dendritic cells derived from human peripheral blood monocytes / 浙江大学学报（英文版）（B辑：生物医学和生物技术）

<p><b>OBJECTIVE</b>This study is aimed at developing a simple and easy way to generate dendritic cells (DCs) from human peripheral blood monocytes (PBMCs) in vitro.</p><p><b>METHODS</b>PBMCs were isolated directly from white blood cell rather than whole blood and purified by patching methods (collecting the attached cell and removing the suspension cell). DCs were then generated by culturing PBMCs for six days with 30 ng/ml recombinant human granulocyte-macrophage stimulating factor (rhGM-CSF) and 20 ng/ml recombinant human interleukin-4 (rhIL-4) in vitro. On the sixth day, TNF-alpha (TNFalpha) 30 ng/ml was added into some DC cultures, which were then incubated for two additional days. The morphology was monitored by light microscopy and transmission electronic microscopy, and the phenotypes were determined by flow cytometry. Autologous mixed leukocyte reactions (MLR) were used to characterize DC function after TNFalpha or lipopolysaccharide (LPS) stimulations for 24 h.</p><p><b>RESULTS</b>After six days of culture, the monocytes developed significant dendritic morphology and a portion of cells expressed CD1a, CD80 and CD86, features of DCs. TNFalpha treatment induced DCs maturation and up-regulation of CD80, CD86 and CD83. Autologous MLR demonstrated that these DCs possess potent T-cell stimulatory capacity.</p><p><b>CONCLUSION</b>This study developed a simple and easy way to generate DCs from PBMCs exposed to rhGM-CSF and rhIL-4. The DCs produced by this method acquired morphologic and antigenic characteristics of DCs.</p>

A comparison of four methods for extraction of human fecal DNA by using real time PCR / 中华检验医学杂志

Objective To compare the relative efficacy and quality of extraction of human fecal DNA using four methods.Methods Real-time PCR were utilized for analysis both quantification and quality of the fecal targeted bacteria(including gut all eubaeterium,Bacteriodes-PrevoteUa group,Bifidobacterium spp Enterobacteriaceae and Enterococcus spp)by using 16s rRNA gene-targeted genus or group-specific primer sets.Results The negative rat of PCR product from method 3(phenol-chloroform plus bead-beating) was about 40%(4/10)by using universal primers,the PCR inhibition disappeared after fecal DNA purified with column.The total fecal 16s rRNA gene copy numbers(per gram of wet weight of feces)as well as the numbers of Bacteriodes-Prevotella group from method 1(QIAamp~DNA stool mini kit)and 4(QIAamp~ DNA stool mini kit combined with bead-beating)was higher significantly than that from method 2(FastDNA ~Kit,Biol01)and 3(P