Clinical Characteristics and Durations of Hospitalized Patients with COVID-19 in Beijing: A Retrospective Cohort Study

Objective:To provide information on clinical characteristics and different durations of COVID-19 and to identify the potential risk factors for longer hospitalization of patients with COVID-19.Methods:In this retrospective study,we enrolled 77 patients(age 52±20 years;44.2%males)with laboratory-confi rmed COVID-19 admitted to Beijing YouAn Hospital between January 21 and February 8,2020.Epidemiological,clinical,and radiological data on admission were collected;complications and outcomes were followed up until February 26,2020.The end point of the study was discharge alive within 2 weeks.Cox proportional-hazards regression was performed to identify risk factors for longer hospitalization.Results:Of 77 patients,there were 34 males(44.2%),24(31.2%)with comorbidities,22(28.6%)with lymphopenia,20(26.0%)with severe COVID-19,and 28(36.4%)with complications.By the end of follow-up,64 patients(83.1%)were discharged home,eight remained in hospital,and fi ve had died.Thirty-six patients(46.8%)were discharged within 14 days and thus reached the study end point,including 34 of the 57 patients with nonsevere COVID-19(59.6%)and two of the 20 patients with severe COVID-19(10%).The overall cumulative probability of the end point was 48.3%.Hospital length of stay and the duration from exposure to discharge for the 64 discharged patients were 13(10-16.5)days and 23(18-24.5)days,respectively.A multivariable stepwise Cox regression model showed that bilateral pneumonia on CT scan,shorter time from illness onset to admission,severity of disease,and lymphopenia were independently associated with longer hospitalization.Conclusions:COVID-19 has a shorter duration of disease and hospital length of stay than severe acute respiratory syndrome.Bilateral pneumonia on CT scan,shorter period from illness onset to admission,lymphopenia,and severity of disease are the risk factors for longer hospitalization of patients with COVID-19.

Liver histological changes in untreated chronic hepatitis B patients in indeterminate phase

BACKGROUND Studies with large size samples on the liver histological changes of indeterminate phase chronic hepatitis B(CHB)patients were not previously conducted.AIM To assess the liver histological changes in the indeterminate phase CHB patients using liver biopsy.METHODS The clinical and laboratory data of 1532 untreated CHB patients were collected,and all patients had least once liver biopsy from January 2015 to December 2021.The significant differences among different phases of CHB infection were compared with t-test,and the risk factors of significant liver histological changes were analyzed by the multivariate logistic regression analysis.RESULTS Among 1532 untreated CHB patients,814(53.13%)patients were in the indeterminate phase.Significant liver histological changes(defined as biopsy score≥G2 and/or≥S2)were found in 488/814(59.95%)CHB patients in the indete-rminate phase.Significant liver histological changes were significant differences among different age,platelets(PLTs),and alanine aminotransferase(ALT)subgroup in indeterminate patient.Multivariate logistic regression analysis indicated that age≥40 years old[adjust odd risk(aOR),1.44;95%confidence interval(CI):1.06-1.97;P=0.02],PLTs≤150×10^(9)/L(aOR,2.99;95%CI:1.85-4.83;P<0.0001),and ALT≥upper limits of normal(aOR,1.48;95%CI:1.08,2.05,P=0.0163)were independent risk factors for significant liver histological changes in CHB patients in the indeterminate phase.CONCLUSION Our results suggested that significant liver histological changes were not rare among the untreated CHB patients in indeterminate phase,and additional strategies are urgently required for the management of these patients.

The default mode network is affected in the early stage of simian immunodeficiency virus infection:a longitudinal study

Acquired immune deficiency syndrome infection can lead to cognitive dysfunction represented by changes in the default mode network.Most recent studies have been cross-sectional and thus have not revealed dynamic changes in the default mode network following acquired immune deficiency syndrome infection and antiretroviral therapy.Specifically,when brain imaging data at only one time point are analyzed,determining the duration at which the default mode network is the most effective following antiretroviral therapy after the occurrence of acquired immune deficiency syndrome.However,because infection times and other factors are often uncertain,longitudinal studies cannot be conducted directly in the clinic.Therefore,in this study,we performed a longitudinal study on the dynamic changes in the default mode network over time in a rhesus monkey model of simian immunodeficiency virus infection.We found marked changes in default mode network connectivity in 11 pairs of regions of interest at baseline and 10 days and 4 weeks after virus inoculation.Significant interactions between treatment and time were observed in the default mode network connectivity of regions of interest pairs area 31/V6.R and area 8/frontal eye field(FEF).L,area 8/FEF.L and caudal temporal parietal occipital area(TPOC).R,and area 31/V6.R and TPOC.L.ART administered 4 weeks after infection not only interrupted the progress of simian immunodeficiency virus infection but also preserved brain function to a large extent.These findings suggest that the default mode network is affected in the early stage of simian immunodeficiency virus infection and that it may serve as a potential biomarker for early changes in brain function and an objective indicator for making early clinical intervention decisions.

A Case of Acute HIV-1 and Monkeypox Coinfection After Condomless Insertive Anal Sex in the Previous 69 Days—Beijing Municipality,China,August-October,2023

Summary What is already known about this topic?The prevalence of monkeypox(mpox)infections is primarily observed among young men who engage in sexual activities with other men,and there is a possibility of sexual transmission.Co-occurring sexually transmitted infections have also been documented.What is added by this report?In this report,we present a case of a patient in China who was simultaneously diagnosed with mpox,and acute human immunodeficiency virus(HIV)infection.

中国六省儿童HIV感染者的流行病学调查

目的了解中国六省儿童HIV感染者的流行病学状况。方法对6个艾滋病高发省份的儿童感染者进行横断面调查,采集患者人口学、感染途径、诊断时间、临床分期、实验室检测等数据。结果共650例患儿入选,男405例,女245例,平均年龄(7.9±3.2)岁,可能感染到确诊的平均时间间隔为(7.1±3.2)年。依次分布于河南570例(87.7%),广西23例(3.5%),云南21例(3.2%),湖北19例(2.9%),安徽10例(1.5%),山西7例(1.1%)。其中母婴传播488例(75.1%),输血及血制品传播102例(15.7%),静脉吸毒传播3例(0.5%)。以采供血为主要传播途径的省份(包括河南、山西、湖北、安徽)和以静脉吸毒为主要传播途径的省份(包括广西、云南)感染者的平均年龄分别为(8.1±3.2)和(5.4±2.2)岁,两组相比差异有显著性(P<0.001)。178例(39.3%,178/453)符合接受抗病毒治疗标准,其中133例(74.7%,133/178)未接受治疗,45例(25.3%,45/178)采用成人药物治疗。结论母婴传播是儿童感染HIV的主要途径,应当加强我国儿童HIV感染者的诊断和治疗工作。

手足口病患儿外周血T淋巴细胞亚群的检测及临床意义

目的检测手足口病患儿外周血淋巴细胞亚群,探讨其免疫功能变化与疾病的关系。方法采用流式细胞仪检测2010年5～10月北京佑安医院感染科病房诊治的119例手足口病患儿急性期的T淋巴细胞亚群,对其中96例患儿咽拭子标本进行反转录-聚合酶链反应(RT-PCR)以确定病原。对不同样本进行独立样本t检验及单因素方差分析。结果手足口病患儿CD3+、CD4+、CD8+绝对数均数随着患儿年龄组的增长而下降。重症病例各T淋巴细胞亚群均数值均低于普通病例(P<0.05),其中重症组CD3+、CD4+、CD8+绝对数分别(1 989.17±1 224.08)×106/L,(1 027.00±771.87)×106/L,(734.00±438.99)×106/L,普通病例组分别为(2 918.08±2 221.74)×106/L,(1 560.38±742.94)×106/L,(1 010.37±687.67)×106/L,CD4+/CD8+差异无统计学意义(P>0.05)。肠道病毒(EV)71阳性与阴性组患儿T淋巴细胞亚群各值比较差异无统计学意义(P>0.05)。结论手足口病患儿存在细胞免疫功能紊乱。

Efficient generation of hepatocyte-like cells from human induced pluripotent stem cells

人的导致的 pluripotent 茎(iPS ) 房间类似于胚胎的茎(ES ) 房间，和罐头强烈地增殖并且区分进许多房间类型。然而，人的 iPS 房间的肝的区别还没被报导了。在这份报告，人的 iPS 房间被导致由一个逐步的协议区分进肝的房间。肝房间标记的表达式和人的 iPS 的肝相关的函数导出房间的房间被监视并且与区分的人的 ES 房间和主要人的 hepatocytes 的相比。在白天 7 点的约 60% 区分的人的 iPS 房间表示了肝的标记 alpha fetoprotein 和白长袍的。在白天 21 点的区分的房间包括白朊 Asecretion，肝糖合成，脲生产和可诱导的细胞色素 P450 活动展出了肝房间功能。肝的标记的表示和 iPS 的肝相关的功能导出房间的肝的房间比得上人的 ES 导出房间的肝的房间的。这些结果证明人的 iPS 房间，类似于人的 ES 房间，能高效地被导致区分房间进象 hepatocyte 一样。

Clinical features and risk factors of acute hepatitis E with severe jaundice

AIM:To compares the clinical features of patients infected with hepatitis E virus(HEV) with or without severe jaundice.In addition,the risk factors for HEV infection with severe jaundice were investigated.METHODS:We enrolled 235 patients with HEV into a cross-sectional study using multi-stage sampling to select the study group.Patients with possible acute hepatitis E showing elevated liver enzyme levels were screened for HEV infection using serologic and molecular tools.HEV infection was documented by HEV antibodies and by the detection of HEV-RNA in serum.We used χ2 analysis,Fisher's exact test,and Student's t test where appropriate in this study.Significant predictors in the univariate analysis were then included in a forward,stepwise multiple logistic regression model.RESULTS:No significant differences in symptoms,alanine aminotransferase,aspartate aminotransferase,al-kaline phosphatase,or hepatitis B virus surface antigen between the two groups were observed.HEV infected patients with severe jaundice had significantly lower peak serum levels of γ-glutamyl-transpeptidase(GGT)(median:170.31 U/L vs 237.96 U/L,P = 0.007),significantly lower ALB levels(33.84 g/L vs 36.89 g/L,P = 0.000),significantly lower acetylcholine esterase(CHE) levels(4500.93 U/L vs 5815.28 U/L,P = 0.000) and significantly higher total bile acid(TBA) levels(275.56 μmol/L vs 147.03 μmol/L,P = 0.000) than those without severe jaundice.The median of the lowest point time tended to be lower in patients with severe jaundice(81.64% vs 96.12%,P = 0.000).HEV infected patients with severe jaundice had a significantly higher viral load(median:134 vs 112,P = 0.025) than those without severe jaundice.HEV infected patients with severe jaundice showed a trend toward longer median hospital stay(38.17 d vs 18.36 d,P = 0.073).Multivariate logistic regression indicated that there were significant differences in age,sex,viral load,GGT,albumin,TBA,CHE,prothrombin index,alcohol overconsumption,and duration of admission between patients infected with acute hepatitis E with and without severe jaundice.CONCLUSION:Acute hepatitis E patients may naturally present with severe jaundice.

Diagnostic value of glypican-3 in serum and liver for primary hepatocellular carcinoma

AIM:To evaluate the diagnostic value of glypican-3(GPC3) in serum and liver for primary hepatocellular carcinoma(HCC).METHODS:Serum levels of GPC3 and α-fetoprotein(AFP) were measured in 75 patients with primary HCC and 32 patients with liver cirrhosis.Expression of GPC3 and AFP in 58 HCC and 12 cirrhotic specimens was detected with immunohistochemical staining.RESULTS:When the cut-off value of serum GPC3 was set at 300 ng/L,its sensitivity and specificity for HCC were 47.0% and 93.5%,respectively.Among the 14 patients with HCC at stage according to the Barcelona Clinic Liver Cancer staging system,the serum GPC3 level was higher than 300 ng/L in 50%(7/14) patients,the serum AFP level was not ≥ 400 μg/L in any patient.Combined serum AFP and GPC3 significantly increased the sensitivity to the diagnosis of HCC.The GPC3 expression was detected in cytoplasm of HCC cells but not in hepatocytes and bile ducts of benign tumors.Among the 58 HCC patients,the GPC3 was expressed in 100%(28/28) patients with their serum AFP level ≥ 400 μg/L,and in 90%(27/30) patients with their AFP level < 400 μg/L,respectively.The GPC3 was weakly or negatively expressed in all paracarcinomatous and cirrhotic tissue samples.AFP positive HCC cells were only found in 1 out of the 58 HCC patients.CONCLUSION:GPC3 protein is a sensitive and specific serum marker for diagnosis of early HCC.Its expression in liver tissues can be used to discriminate tumor cells from benign hepatic cells.

Clinical features of adverse reactions associated with telbivudine

AIM: To analyze the clinical features and risk factors of adverse reactions associated with telbivudine. METHODS: Clinical data were collected from cases that presented with serious adverse reactions to telbivudine. We analyzed general information and medicine status, clinical features, results of examination, and misdiagnosis. RESULTS: Out of 105 patients who were treated with telbivudine for hepatitis B in an outpatient department from January, 2007 to January, 2008, five presented with serious adverse drug reactions. Most of these five patients had used other nucleoside analogues in the past. Four were treated with a combination of telbivudine and interferon or another nucleoside analogue, while the other received an increased dose of telbivudine. The main adverse reactions were myalgia and general weakness. This was accompanied by cardiac arrhythmia in one patient, and nervous symptoms in three. Serum creatine kinase was elevated. The rate of misdiagnosis was high. CONCLUSION: The adverse reactions were related to telbivudine, but the biological mechanism of the reactions is not yet clear. Combination therapy with interferon or another nucleoside analogue and a high dose may increase the risk of adverse reactions.

Dual gRNAs guided CRISPR/Cas9 system inhibits hepatitis B virus replication

AIM: To screen and investigate the effective g RNAs against hepatitis B virus(HBV) of genotypes A-D.METHODS: A total of 15 g RNAs against HBV of genotypes A-D were designed. Eleven combinations of two above g RNAs(dual-g RNAs) covering the regulatory region of HBV were chosen. The efficiency of each g RNA and 11 dual-g RNAs on the suppression of HBV(genotypes A-D) replication was examined by the measurement of HBV surface antigen(HBs Ag) or e antigen(HBe Ag) in the culture supernatant. The destruction of HBV-expressing vector was examined in Hu H7 cells co-transfected with dual-g RNAs and HBVexpressing vector using polymerase chain reaction(PCR) and sequencing method, and the destruction of ccc DNAwas examined in Hep AD38 cells using KCl precipitation, plasmid-safe ATP-dependent DNase(PSAD) digestion, rolling circle amplification and quantitative PCR combined method. The cytotoxicity of these g RNAs was assessed by a mitochondrial tetrazolium assay.RESULTS: All of g RNAs could significantly reduce HBs Ag or HBe Ag production in the culture supernatant, which was dependent on the region in which g RNA against. All of dual g RNAs could efficiently suppress HBs Ag and/or HBe Ag production for HBV of genotypes A-D, and the efficacy of dual g RNAs in suppressing HBs Ag and/or HBe Ag production was significantly increased when compared to the single g RNA used alone. Furthermore, by PCR direct sequencing we confirmed that these dual g RNAs could specifically destroy HBV expressing template by removing the fragment between the cleavage sites of the two used g RNAs. Most importantly, g RNA-5 and g RNA-12 combination not only could efficiently suppressing HBs Ag and/or HBe Ag production, but also destroy the ccc DNA reservoirs in Hep AD38 cells.CONCLUSION: These results suggested that CRISPR/Cas9 system could efficiently destroy HBV expressing templates(genotypes A-D) without apparent cytotoxicity. It may be a potential approach for eradication of persistent HBV ccc DNA in chronic HBV infection patients.

Prognostic value of M30/M65 for outcome of hepatitis B virus-related acute-on-chronic liver failure

AIM:To determine the prognostic value of circulating indicators of cell death in acute-on-chronic liver failure(ACLF)patients with chronic hepatitis B virus(HBV)infection as the single etiology.METHODS:Full length and caspase cleaved cytokeratin 18(detected as M65 and M30 antigens)represent circulating indicators of necrosis and apoptosis.M65and M30 were identified by enzyme-linked immunosorbent assay in 169 subjects including healthy controls(n=33),patients with chronic hepatitis B(CHB,n=55)and patients with ACLF(n=81).According to the 3-mo survival period,ACLF patients were defined as having spontaneous recovery(n=33)and non-spontaneous recovery which included deceased patients and those who required liver transplantation(n=48).RESULTS:Both biomarker levels significantly increased gradually as liver disease progressed(for M65:P<0.001 for all;for M30:control vs CHB,P=0.072;others:P<0.001 for all).In contrast,the M30/M65 ratio was significantly higher in controls compared with CHB patients(P=0.010)or ACLF patients(P<0.001).In addition,the area under receiver operating characteristic curve(AUC)analysis demonstrated that both biomarkers had diagnostic value(AUC≥0.80)in identifying ACLF from CHB patients.Interestingly,it is worth noting that the M30/M65 ratio was significantly different between spontaneous and non-spontaneous recovery in ACLF patients(P=0.032).The prognostic value of the M30/M65 ratio was compared with the Model for End-Stage Liver Disease(MELD)and Child-Pugh scores at the 3-mo survival period,the AUC of the M30/M65ratio was 0.66 with a sensitivity of 52.9%and the highest specificity of 92.6%(MELD:AUC=0.71;sensitivity,79.4%;specificity,63.0%;Child-Pugh:AUC=0.77;sensitivity,61.8%;specificity,88.9%).CONCLUSION:M65 and M30 are strongly associated with liver disease severity.The M30/M65 ratio may be a potential prognostic marker for spontaneous recovery in patients with HBV-related ACLF.

Prevalence of porcine endogenous retrovirus in Chinese pig breeds and in patients treated with a porcine liver cell-based bioreactor

AIM: To determine the prevalence of porcine endogenous retrovirus (PERV) in various pig breeds raised in China including Chinese experimental mini-pigs by PERV-reverse transcriptase (PERV-RT enzyme). Moreover, the potential for infection of PERV was investigated in patients treated with a bioreactor based on porcine liver cells (n = 3).METHODS: Pig serum, liver and musde cell-free supematants were collected from various Chinese pig breeds. Porcine hepatocytes were isolated with a two-step perfusion method. Three patients with acute or chronic liver failure were treated with a bioartificial liver support system (BALSS) for 8-12 h and serum samples were collected from the patients before, immediately after and 30 d after treatment.RESULTS: The activities of PERV-RT enzyme in pig liver and muscle cell-free supernatants were higher than in normal human controls. PERV-TR enzyme activity did not increase in patients before and after 1 mo of treatment.PERV-RT activities were not significantly different when compared with pre-treatment group (1.544±0.155576), the post-treatment groups (1.501±0.053507, 1.461±0.033808 and 1.6006667±0.01963 for 0, 14 and 30 d post-treatment,respectively, P>0.05), and normal control group (1.440±1.0641, P>0.05). RT enzyme activity in Chinese experimental mini-pigs was higher than in normal human control group (1.440±1.0641 U/mL, P<0.05), and not significantly different (P>0.05) when compared with the pig breeds except in the muscle supernatants. All the samples including muscle and liver cell supernatants from the Chinese mini-experimental pigs and the four domestic Chinese pig breeds contained PERVs.CONCLUSION: These results suggest that the risk of PERV infection through BALSS containing porcine liver cells without immunosuppressants may be quite low. Although there were PERVs in Chinese experimental mini-pigs and porcine liver cell culture suspensions, we did not find any evidence of persistent PERV infection in patients treated with this porcine hepatocyte-based bioartificial liver.

A more consistent intraluminal rhesus monkey model of ischemic stroke

Endovascular surgery is advantageous in experimentally induced ischemic stroke because it causes fewer cranial traumatic lesions than invasive surgery and can closely mimic the pathophysiology in stroke patients. However, the outcomes are highly variable, which limits the accuracy of evaluations of ischemic stroke studies. In this study, eight healthy adult rhesus monkeys were randomized into two groups with four monkeys in each group： middle cerebral artery occlusion at origin segment （M1） and middle cerebral artery occlusion at M2 segment. The blood flow in the middle cerebral artery was blocked completely for 2 hours using the endovascular microcoil placement technique （1 mm × 10 cm） （undetachable）, to establish a model of cerebral ischemia. The microcoil was withdrawn and the middle cerebral artery blood flow was restored. A reversible middle cerebral artery occlusion model was identified by hematoxylin-eosin staining, digital subtraction angiography, magnetic resonance angiography, magnetic resonance imaging, and neurological evaluation. The results showed that the middle cerebral artery occlusion model was successfully established in eight adult healthy rhesus monkeys, and ischemic lesions were apparent in the brain tissue of rhesus monkeys at 24 hours after occlusion. The rhesus monkeys had symptoms of neurological deficits. Compared with the M1 occlusion group, the M2 occlusion group had lower infarction volume and higher neurological scores. These experimental findings indicate that reversible middle cerebral artery occlusion can be produced with the endovascular microcoil technique in rhesus monkeys. The M2 occluded model had less infarction and less neurological impairment, which offers the potential for application in the field of brain injury research.

Antiviral treatment for chronic hepatitis B: Safety, effectiveness, and prognosis

The goal of chronic hepatitis B (CHB) therapy is to improve the patient prognosis through the sustained inhibition of viral replication. However, due to the uncertainty and potentially unlimited duration of the treatment course, nucleus(t)ide analogue (NA) resistance and safety, financial costs and patient compliance, different endpoints of antiviral treatment have been proposed in CHB prevention and treatment guidelines. Different treatment endpoints are closely associated with the safety of drug withdrawal and improvements in prognosis. Antiviral treatment suppresses HBV DNA replication, drug withdrawal leads to relapse, and long-term treatment causes drug safety and resistance issues. Although hepatitis B e antigen seroconversion based on HBV DNA inhibition is considered as “a satisfactory endpoint”, drug withdrawal still leads to high relapse rates. Hepatitis B surface antigen (HBsAg) clearance is the “ideal endpoint” in terms of the safety of drug withdrawal and improvements in prognosis. However, the HBsAg clearance rate is low using the conventional single drug treatment and fixed course regimens. Recently, the application of an “optimized antiviral treatment strategy” has improved the HBsAg clearance rate, and make an “ideal endpoint” possible. This article reviews the different antiviral treatment endpoints in terms of the safety of drug withdrawal, improvements in prognosis and relevant advances.

C-arm Lipiodol CT in transcatheter arterial chemoembolization for small hepatocellular carcinoma

AIM:To investigate the value of C-arm Lipiodol computed tomography(CT) for intra-procedural hepatocellular carcinoma(HCC) lesion detection during transcatheter arterial chemoembolization(TACE).METHODS:Forty patients(37 male,3 female;mean age,52.6 ± 12.5 years,age range:25-82 years) diagnosed with HCC were enrolled in this study.All patients underwent 64-slice CT 1-2 wk before TACE.During the procedure,hepatic angiography was performed first.Following diagnostic embolization with Lipiodol injected into the hepatic artery,a C-arm CT scan was immediately conducted(C-arm Lipiodol CT).If new HCC lesions were confirmed,gelfoam particles were super-selectively injected into the tumor-nourishing blood vessel.A Lipiodol CT scan was performed 7-14 d after TACE.All images acquired from 64-slice CT,digital subtraction angiography(DSA),C-arm Lipiodol CT and Lipiodol CT were retrospectively reviewed by four radiologists and the number of detected lesions in each examination was counted,respectively.The results of Lipiodol CT were taken as the diagnostic reference.Alpha-fetoprotein values were examined both before and after TACE.This study only takes into account the lesions that were not found or were considered suspicious on 64-slice CT before TACE.RESULTS:Preprocedural 64-slice CT detected a total of 13 suspicious lesions in the 40 patients.DSA detected ten definite and four suspicious lesions.C-arm Lipiodol CT detected 71 lesions in total and Lipiodol CT confirmed 67 lesions with a diameter range of 3-12 mm.Four false-positive lesions,which were detected by C-arm Lipiodol CT,were considered to be hepatic artery-portal vein fistulas.The average alpha-fetoprotein values before and after TACE were significantly different(452.3 ± 192.6 ng/m L vs 223.8 ± 93.2 ng/m L;P = 0.039).CONCLUSION:C-arm Lipiodol CT has a higher diagnostic sensitivity for small HCC lesions.This technique may help physicians make intraproceduraldecisions to provide patients with earlier treatment.

Autophagy in anti-apoptotic effect of augmenter of liver regeneration in HepG2 cells

AIM:To investigate the role of autophagy in the antiapoptotic effect of augmenter of liver regeneration(ALR).METHODS:Autophagy was induced through serum deprivation.An ALR-expressing plasmid was transfected into HepG2 cells,and autophagic flux was determined using fluorescence microscopy,electron microscopy,Western blot and quantitative polymerase chain reaction(q PCR) assays.After ALR-expressing plasmid transfection,an autophagy inhibitor [3-methyladenine(3-MA)] was added to HepG2 cells,and apoptosis was observed using fluorescence microscopy and flow cytometry.RESULTS:Autophagy was activated in HepG2 cells,peaking at 24 h after serum deprivation.Microtubuleassociated protein light chain three-II levels were higher in HepG2 cells treated with ALR than in control cells,fluorescence microscopy,electron microscopy and q PCR studies showed the similar trend,and p62 levels showed the opposite trend,which indicated that ALR may play an important role in increasing autophagy flux.The numbers of apoptotic cells were substantially higher in HepG2 cells treated with both ALR and 3-MA than in cells treated with ALR alone.Therefore,the protective effect of ALR was significantly attenuated or abolished when autophagy was inhibited,indicating that the anti-apoptotic effect of ALR may be related to autophagy.CONCLUSION:ALR protects cells from apoptosis partly through increased autophagy in HepG2 cells and may be valuable as a new therapeutic treatment for liver disease.

Effect of individualized therapy for AIDS patients with cytomegalovirus retinitis in intravitreal ganciclovir injections

The effect of intravitreal ganciclovir injection combined with intravenous infusion on acquired immune deficiency syndrome(AIDS) patients with cytomegalovirus retinitis(CMVR) was investigated. A total of 32 eyes in 23 AIDS patients diagnosed as CMVR from 2017 to 2018 were included in the retrospective study. All patients underwent induction therapy by using intravenous drip of the anticytomegalovirus(CMV) agent ganciclovir(5 mg/kg q12 h) combined with intravitreal ganciclovir injection(3 mg/time, 2 times/wk). The visual acuity, fundus photographs, lesion location, and number of intravitreal injections were observed preoperatively and postoperatively. Totally 14 eyes were cured during induction therapy. The number of injections [4.13(2 to 6)] in CMVR patients with peripherally fundus lesions were significantly lower than those with central lesions (4.89(2 to 6))The individualized therapy of intravitreal ganciclovir injections for AIDS patients with CMVR can effectively reduce the numbers of intravitreal injections.

Incomplete radiofrequency ablation promotes the development of CD133+cancer stem cells in hepatocellular carcinoma cell line HepG2 via inducing SOX9 expression

Background: Cancer stem cells(CSCs) accelerate the growth of hepatocellular carcinoma(HCC) residual after incomplete radiofrequency ablation(In-RFA). The present study aimed to detect the effects of In-RFA on stemness transcription factors(STFs) expression which are important for the production and function of CSCs, and to find which STFs promote HCC stemness after In-RFA. Methods: HepG2 cells were used for in vitro and in vivo studies. Flow cytometry and sphere-formation assays were used to detect the level and function of CD133~+ CSCs in the models. PCR array and ELISA were applied to analyze the altered expression of 84 STFs in CD133~+ CSCs in two models. Specific lentiviral shRNA was used to knockdown STFs expression, followed by detecting In-RFA’s effects on the levels and function of CD133~+ CSCs. Results: In-RFA was identified to induce CD133~+ CSCs and increase their tumorigenesis ability in vitro and in vivo. The mRNA levels of 84 STFs in CD133~+ CSCs were detected by PCR array, showing that 15 and 22 STFs were up-regulated in two models, respectively. Meanwhile, the mRNA levels of seven common STFs were up-regulated in both models. ELISA assay demonstrated that only the protein of sex determining region Y-box 9(SOX9) was up-regulated in both models, the protein levels of the other 6 common STFs did not increase in both models. Finally, SOX9 was identified to play an important role in inducing, maintaining stemness and promoting tumorigenesis ability of CD133~+ CSCs in both models. Conclusion: In-RFA-induced SOX9 stimulates CD133~+ CSCs proliferation and increases their tumorigenesis ability, suggesting that SOX9 may be a good target for HCC treatment.

Curcumin ameliorates hippocampal neuron damage induced by human immunodeficiency virus-1

Our previous studies have shown that infection with the gp120 V3 loop can cause human immunodeficiency virus-1 associated neurocognitive disorders. Curcumin has been shown to improve these effects to some degree, but the precise mechanisms remain unknown. The present study analyzed the neuroprotective effect and mechanism of curcumin in relation to hippocampal neurons. Results showed that 1 nmol/L gp120 V3 loop suppressed the growth of synapses. After administration of 1 !umol/L curcumin, synaptic growth improved. Curcumin is neuroprotective against gp120 V3 loop-induced neuronal damage by inhibiting the activation of L-type calcium currents, relieving intracellular Ca^2＋ overload, promoting Bcl-2 expression, and inhibiting Bax activation. The effect of curcumin was identical to nimodipine, suggesting that curcumin has the same neuroprotective effects against gp120 V3 loop-induced neuronal damage.