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OBJECTIVE@#To explore the value of in situ amniocyte culture for prenatal diagnosis.@*METHODS@#2716 amniotic fluid samples were cultured in situ on slides. After the culture, the slides were stained, photographed and analyzed.@*RESULTS@#All samples were successfully analyzed, with the success rates for primary culture and subculture being 98.42% and 1.58%, respectively. 224 samples (8.25%) were detected with chromosomal aberrations, which included 125 cases with trisomy 21, 31 with trisomy 18, 3 with trisomy 13, 4 with 45,X, 17 with 47,XXY, 5 with 47,XYY, 1 with 48,XXY,+18, 1 with 48,XXYY, 26 with structural chromosomal aberrations, and 11 with mosaicisms for aneuploidies.@*CONCLUSION@#In situ amniocyte culture is stable and has a high success rate, and is capable of identifying true and false mosaicisms, which can improve the accuracy of prenatal diagnosis.
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Objective:To compare the efficacy of the combination of abidol, lopinavir/ritonavir plus recombinant interferon α-2b (rIFNα-2b) and the combination of lopinavir/ritonavir plus rIFNα-2b for patients with COVID-19 in Zhejiang province.Methods:A multicenter prospective study was carried out to compare the efficacy of triple combination antiviral therapy and dual combination antiviral therapy in 15 medical institutions of Zhejiang province during January 22 to February 16, 2020. All patients were treated with rIFNα-2b (5 million U, 2 times/d) aerosol inhalation, in addition 196 patients were treated with abidol (200 mg, 3 times/d) + lopinavir/ritonavir (2 tablets, 1 time/12 h) (triple combination group) and 41 patients were treated with lopinavir/ritonavir (2 tablets, 1 time/12 h) (dual combination group). The patients who received triple combination antiviral therapy were further divided into three subgroups: <48 h, 3-5 d and >5 d according the time from the symptom onset to medication starting. The therapeutic efficacy was compared between triple combination group and dual combination group, and compared among 3 subgroups of patients receiving triple combination antiviral therapy. SPSS 17.0 software was used to analyze the data.Results:The virus nucleic acid-negative conversion time in respiratory tract specimens was (12.2±4.7) d in the triple combination group, which was shorter than that in the dual combination group [(15.0±5.0) d] ( t=6.159, P<0.01). The length of hospital stay in the triple combination group [12.0 (9.0, 17.0) d] was also shorter than that in the dual combination group [15.0 (10.0, 18.0) d] ( H=2.073, P<0.05). Compared with the antiviral treatment which was started within after the symptom onset of in the triple combination group, the time from the symptom onset to the viral negative conversion was 13.0 (10.0, 17.0), 17.0 (13.0, 22.0) and 21.0 (18.0, 24.0) d in subgroups of 48 h, 3-5 d and >5 d, respectively ( Z=32.983, P<0.01), while the time from antiviral therapy to viral negative conversion was (11.8±3.9), (13.5±5.1) and (11.2±4.3) d, respectively( Z=6.722, P<0.05). Conclusions:The triple combination antiviral therapy of abidol, lopinavir/litonavir and rIFNα-2b shows shorter viral shedding time and shorter hospitalization time, compared with the dual combination antiviral therapy; and the earlier starting triple combination antiviral therapy will result in better antiviral efficacy.
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Objective@#Comparing the benefit of Abidor, lopinavir/ritonavir and recombinant interferon α-2b triple combination antiviral therapy and lopinavir/ritonavir and interferon dual combination antiviral therapy to hospitalized novel coronavirus pneumonia 2019 in Zhejiang province.@*Methods@#A multi-center prospective study was carried out to compare the effect of triple combination antiviral therapy with dual combination antiviral therapy in 15 medical institutions of Zhejiang Province. All patients were treated with recombinant interferon α-2b (5 million U, 2 times/d) aerosol inhalation. 196 patients were treated with abidol (200 mg, 3 times/d) + lopinavir / ritonavir (2 tablets, 1 time/12 h) as the triple combination antiviral treatment group. 41 patients were treated with lopinavir / ritonavir (2 tablets, 1 time/12 h) as the dual combination antiviral treatment group. The patients who received triple combination antiviral therapy were divided into three groups: within 48 hours, 3-5 days and > 5 days after the symptom onset. To explore the therapeutic effects of triple combination antiviral drugs and dual combination antiviral drugs, as well as triple combination antiviral drugs with different antiviral initiate time. SPSS17.0 software was used to analyze the data.@*Results@#The time of virus nucleic acid turning negative was (12.2 ± 4.7) days in the triple combination antiviral drug group, which was shorter than that in the dual combination antiviral drug group [(15.0 ± 5.0) days] (t = 6.159, P < 0.01 ). The length of hospital stay [12 (9, 17) d] in the triple combination antiviral drug group was also shorter than that in the dual combination antiviral drug group [15 (10, 18) d] (H = 2.073, P < 0.05). Comparing the antiviral treatment which was started within 48 hours, 3-5 days and > 5 days after the symptom onset of triple combination antiviral drug group, the time from the symptom onset to the negative of viral shedding was 13 (10,16.8), 17 (13,22) and 21 (18-24) days respectively (Z = 32.983, P < 0.01), and the time from antiviral therapy to the negative of viral shedding was (11.8±3.9) , (13.5±5.1) and (11.2±4.3) d. The differences among the three groups were statistically significant (Z=32.983 and 6.722, P<0.01 or<0.05).@*Conclusions@#The triple combination antiviral therapy of Abidor, Lopinavir/Litonavir and recombinant interferon α-2b showed shorter viral shedding time and hospitalization time compared with the dual combination antiviral therapy. The earlier the time to initiate triple antiviral treatment, the shorter the time of virus shedding.
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Objective@#To investigate the effect of hepatitis B virus (HBV) X gene integration on expression of zinc finger protein ZBTB20 in chronic hepatitis B (CHB) patients complicated with hepatocellular carcinoma (HCC).@*Methods@#Eighteen CHB patients complicated with HCC who underwent surgical treatment in Taizhou Enze Medical Center Enze Hospital and Taizhou Central Hospital from July 2015 to June 2017 were enrolled. Samples of carcinoma tissue, para-carcinoma tissue and corresponding normal liver tissue were collected from each case. DNA was extracted from three kinds of tissue samples. HBV-Alu-polymerase chain reaction (PCR) was used to amplify the integrated HBVX fragments and their bilateral flanking sequences in human genomic DNA. The integrated HBV fragments were determined by PCR products sequencing. Protein was extracted from three kinds of tissue samples.The level of expression of ZBTB20 was detected by protein imprinting. Statistical analysis was performed using t test, analysis of variance, and χ2 test.@*Results@#Among the 18 CHB patients complicated with HCC, integration of HBVX gene was found in 13 carcinoma tissue samples, 16 para-carcinoma tissue samples and 9 corresponding normal liver tissue samples. The difference was statistically significant (χ2=6.353, P=0.037). Of the 18 patients, the protein expressions of ZBTB20 in carcinoma tissue, para-carcinoma tissue and corresponding normal tissue were (50.14±11.25)%, (40.71±7.17)% and (39.06±5.17)%, respectively, which was statistically different (F=9.420, P<0.01). HBVX gene integration was detected at ZBTB20 locus in five patients. The expression levels of ZBTB20 in patients with HBVX gene integration at this locus in carcinoma tissue, para-carcinoma tissue and corresponding, normal liver tissue were all significantly lower than those in patients without HBVX gene integration (carcinoma tissue (37.37±10.30)% vs (55.06±7.06)%, para-carcinoma tissue (32.06±2.61)% vs (44.04±5.24)%, corresponding normal tissue (34.66±5.59)% vs (40.76±4.04)%, t=4.205, 4.821 and 2.589, respectively, all P<0.05).@*Conclusions@#Incidence of HBVX integration in para-carcinoma tissue is higher than that in carcinoma tissue in CHB patients complicated with HCC.The expression level of ZBTB20 in carcinoma tissue is higher than that in para-carcinoma tissue. Integration of HBVX gene at ZBTB20 locus may decreases the expression of ZBTB20.
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Objective To assess the diagnostic value of serum soluble mannose receptor (sMR) for hepatic fibrosis in patients with chronic hepatitis B (CHB).Methods Fifty patients with CHB undergoing liver biopsy in the Department of Infectious Diseases , Taizhou Hospital of Zhejiang Province from November 2016 to October 2018 were enrolled, including 28 males and 22 females.According to the stage of liver fibrosis, there were 15 cases without fibrosis (S0 group), 12 cases of mild fibrosis (S1-2 group), and 15 cases of moderate-severe fibrosis ( S3-4 group).Twenty healthy subjects (12 males and 8 females) were recruited as controls.Enzyme linked immunosorbent assay (ELISA) was used to detect the serum hyaluronic acid (HA), laminin (LN), procollagen type ⅢN-terminal peptide (PⅢP), collagen type IV (CIV) and sMR in all groups.One-way ANOVA, Spearman correlation analysis and receiver operating characteristic (ROC) curve were used to analyze the data.Results The serum levels of sMR, HA, LN, CIV and PⅢP in S3-4 group were significantly higher than those in S 0 group ( t＝10.20, 4.69, 8.94, 2.35 and 4.34, respectively; all P＜0.05) and S1-2 group (t＝5.77, 4.23, 7.88, 2.71 and 3.43, respectively; all P＜0.05); and serum sMR level in S1-2 group was higher than that in S0 group ( t ＝6.23, P ＜0.05). Spearman rank correlation demonstrated that serum sMR level was positively correlated with the degree of liver fibrosis (r＝0.860, P＜0.01).ROC curve analysis showed that when 228.69 ng／mL was taken as cut-off value of sMR, its specificity and sensitivity for diagnosis of hepatic fibrosis were 93.3%and 88.6%, respectively.The diagnostic efficacy of sMR was significantly better than that of HA , LN, CIV and PⅢP (Z＝3.179, 3.467, 5.241 and 3.567, respectively; all P＜0.05).When 345.80 ng／mL was taken as cut-off value of sMR, the specificity and sensitivity for diagnosis of moderate to severe hepatic fibrosis were 85.7%and 86.7%, respectively; and its diagnostic efficacy was better than that of HA , CIV and PⅢP (Z＝2.253, 2.475 and 2.092, all P ＜0.05).Conclusion Serum sMR level is associated with the progression of liver fibrosis, it may be used as a new serological marker for non-invasive assessment of liver fibrosis.
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Objective To investigate the effect of hepatitis B virus (HBV) X gene integration on expression of zinc finger protein ZBTB20 in chronic hepatitis B (CHB) patients complicated with hepatocellular carcinoma (HCC).Methods Eighteen CHB patients complicated with HCC who underwent surgical treatment in Taizhou Enze Medical Center Enze Hospital and Taizhou Central Hospital from July 2015 to June 2017 were enrolled.Samples of carcinoma tissue,para-carcinoma tissue and corresponding normal liver tissue were collected from each case.DNA was extracted from three kinds of tissue samples.HBV-Alu-polymerase chain reaction (PCR) was used to amplify the integrated HBVX fragments and their bilateral flanking sequences in human genomic DNA.The integrated HBV fragments were determined by PCR products sequencing.Protein was extracted from three kinds of tissue samples.The level of expression of ZBTB20 was detected by protein imprinting.Statistical analysis was performed using t test,analysis of variance,and x2 test.Results Among the 18 CHB patients complicated with HCC,integration of HBVX gene was found in 13 carcinoma tissue samples,16 para-carcinoma tissue samples and 9 corresponding normal liver tissue samples.The difference was statistically significant (x2 =6.353,P =0.037).Of the 18 patients,the protein expressions of ZBTB20 in carcinoma tissue,para-carcinoma tissue and corresponding normal tissue were (50.14 ± 11.25)％,(40.71±7.17) ％ and (39.06 ± 5.17) ％,respectively,which was statistically different (F =9.420,P ＜ 0.01).HBVX gene integration was detected at ZBTB20 locus in five patients.The expression levels of ZBTB20 in patients with HBVX gene integration at this locus in carcinoma tissue,para-carcinoma tissue and corresponding,normal liver tissue were all significantly lower than those in patients without HBVX gene integration (carcinoma tissue (37.37±10.30)％ vs (55.06 ± 7.06)％,para-carcinoma tissue (32.06 ± 2.61)％ vs (44.04 ± 5.24)％,corresponding normal tissue (34.66 ±5.59)％ vs (40.76 ±4.04)％,t--4.205,4.821 and 2.589,respectively,all P ＜0.05).Conclusions Incidence of HBVX integration in para-carcinoma tissue is higher than that in carcinoma tissue in CHB patients complicated with HCC.The expression level of ZBTB20 in carcinoma tissue is higher than that in para-carcinoma tissue.Integration of HBVX gene at ZBTB20 locus may decreases the expression of ZBTB20.
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Objective To observe the curative effect of entecavir combined with glucocorticoid in early stage hepatitis B liver failure. Methods The clinical data of 100 patients with early stage hepatitis B liver failure from February 2012 to February 2017 were retrospectively analyzed. Among them, 50 patients were treated with entecavir combined with glucocorticoid (treatment group), and 50 patients were treated with entecavir (control group). The alanine aminotransferase (ALT), total bilirubin (TBil), total cholesterol (TC), prothrombin activity (PTA), hepatitis B virus (HBV)-DNA, tumor necrosis factor (TNF)-α, interleukin (IL)-6 and IL-10 levels were compared between 2 groups. Results The ALT, TBil, TC and PTA levels 1, 2, 4 and 8 weeks after treatment in treatment group were significantly lower than those in control group, and there were statistical differences (P<0.05); there was no statistical difference in HBV-DNA between 2 groups (P>0.05). The TNF-α, IL-6 and IL-10 levels 4 weeks after treatment in treatment group were significantly lower than those in control group: (2.5 ± 0.5) μg/L vs. (3.2 ± 0.7) μg/L, (16.8 ± 2.2) μg/L vs. (22.1 ± 2.4) μg/L and (7.3 ± 1.2) μg/L vs. (9.1 ± 1.8) μg/L, and there were statistical differences (P<0.05). Eight weeks after treatment, the effective rate and survival rate in treatment group were significantly higher than those in control group: 80％ (40/50) vs. 54％ (27/50) and 94％ (47/50) vs. 78％ (39/50), and there were statistical differences (P<0.01 or <0.05).In the treatment group, there was no serious adverse reaction during glucocorticoid therapy,and the patients recovered after withdrawal or reduction of glucocorticoid. Conclusions Entecavir combined with glucocorticoid has a significant effect on patients with early stage hepatitis B liver failure. It could improve survival rate, and the adverse effects of glucocorticoid are less.
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Objective To investigate the effect of alprostadil injection on serum tumor necrosis factor ( TNF-α) ,β2-microglobulin (β2-MG) and soluble interleukin 2 receptor (sIL-2R) levels in adjunctive treatment of patients of severe hepatitis with liver and kidney syndrome.Methods A total of 60 patients of severe hepatitis with liver and kidney syndrome were collected and randomly divided into experimental group and control group with 30 cases in each group.The two groups were treated by conventional therapy, patients in the control group were treated by continuous renal replacement therapy, patients in the experimental group were treated on the basis of the control group with alprostadil injection in adjuvant therapy for 3 weeks.The serum biochemistry and liver and kidney function and clinical curative effect were detected.Results Compared with the control group, the serum TNF-α, β2-MG and sIL-2R levels of the experimental group were lower (P<0.05); alanine aminotransferase (ALT), aspartate aminotransferase (AST) and total bilirnbin (TBiL) levels were lower (P<0.05), albumin (Alb) level and prothrombin time activity (PTA) were higher (P<0.05), serum creatinine (Scr), urea nitrogen (BUN), CysC and 24h urine protein were lower (P<0.05), 24h urine volume was higher (P<0.05); efficacy in experimental group was better than control group (Z=-2.321,P=0.020).Conclusion Alprostadil injection in adjunctive treatment of severe hepatitis with liver and kidney syndrome patients has significant effect and high safety, could significantly reduce the serum TNF-α, β2-MG and sIL-2R levels.
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Objective To evaluate the application value of high-risk human papillomavirus (HR-HPV)DNA de-tection and ThinPrep liquid-based cytology testing(TCT)in cervical disease screening.Methods Cervical specimens of women with cervical lesions in a hospital between October 2012 and December 2013 were taken and performed hu-man papillomavirus DNA genotyping (HPV DNA)and ThinPrep liquid-based cytology testing(TCT).Positive pa-tients were performed colposcopy pathological detection.HPV DNA positive rates among different TCT groups, and different cervical lesion groups were compared,the sensitivity and specificity of TCT and HPV DNA detection, as well as differences between separate and joint detection were also compared.Results The positive rate of HPV DNA was 28.07% (1 045/3 723),most were HR-HPV (21 .57%,n=803),the major HR-HPV genotypes were HPV 16,58,52,and 18 type.HR-HPV positive rates were statistically different among different age groups(χ2 =31 .74,P tection was as a gold standard ,the sensitivity of HR-HPV DNA and TCT was 90.44%(265/293)and 85.32%(250/293)respectively,the sensitivity of joint HR-HPV DNA detection and TCT was 95.90%.In positive patho-logical group,the detection rate of TCT and HR-HPV DNA was 85.32% and 90.44%,respectively,joint detec-tion rate was 95.90%,the difference among three groups was significant (χ2 =18.185,P <0.001).Joint detection rate was higher than separate detection rate of TCT or HPV DNA.Conclusion HPV DNA detection is a useful supplement for cervical cancer screening,HPV DNA detection combined with TCT can reduce the misdiagnosis rate.
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Objective To observe the curative effect of methylprednisolone combined with entecavir in treatment of hepatitis B virus (HBV) related early stage liver failure.Methods One hundred and twenty-six patients with HBV related early stage liver failure were divided into treatment group (68 cases) and control group (58 cases) by random digits table method.The patients in 2 groups were given conventional hepatinica treatment and entecavir antiviral treatment,but the patients in treatment group were added methylprednisolone and pantoprazole.The alanine aminotransferase (ALT),total bilirubin (TBil),albumin,prothrombin time (PT),HBV-DNA,tumor necrosis factor (TNF)-α,interleukin (IL)-6 levels were compared between 2 groups,and the adverse reaction of methylprednisolone was observed.Results The ALT,TBil,PT and albumin levels after the first,second,fourth,sixth and eighth week of treatment in treatment group were significantly better than those in control group,and there were statistical differences (P ＜ 0.05).There was no statistical difference in HBV-DNA between 2 groups (P ＞ 0.05).The TNF-α and IL-6 levels after the first and second week of treatment in treatment group were (4.13 ± 1.25) and (1.98 ± 0.67) p g/L,(3.21 ± 0.75)and (1.23 ± 0.29) μ g/L,and in control groups were (5.89 ± 1.78) and (3.67 ± 0.87)μ g/L,(4.12 ± 0.88) and (2.68 ± 0.81) μ g/L.The TNF-α and IL-6 levels in treatment group were significantly lower than those in control group,and there were statistical differences (P ＜ 0.05).The effective rate in treatment group (79.41％,54168) was significantly higher than that in control group (51.72％,30/58),the fatality rate in treatment group (2.94％,2/68) was significantly lower than that in control group (24.14％,14/58),and there were statistical differences (P ＜ 0.05).The adverse reaction of methylprednisolone in treatment group was not found.Conclusion The methylprednisolone combined with entecavir can improve liver function and survival rate in patients with HBV related early stage liver failure,and adverse reaction of methylprednisolone is rare.
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Objective To express VP1recombinant protein of Echovirus 30 (ECHO30) in E.coli BL21(DE3) and to develop an enzyme-linked immunoassay (EIA) based on the recombinant antigen for detecting antibodies to ECHO30.Methods The target VP1gene was amplified by reverse transcription-polymerase chain reaction (RT-PCR).The PCR products of the gene were inserted into pET44a vector,and then expressed in E.coli BL21( DE3 ).The purified recombinant protein was used for the development of EIA.Results The molecular weight of the recombinant protein was 95 000,and the antigenicity of which was identified by Western blot and EIA.Conclusion The recombinant protein VP1has been successfully expressed and purified,which can be used as diagnostic antigen.
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Objective To review the diagnosis and treatment of hepatic veno-occlusive disease(HVOD)induced by sedum aizoon in HBsAg positive patients. Methods Clinical data of 35 HBsAg positive cases who took sedum aizoon decoction and developed HVOD were collected, the clinical manifestation, imaging examination, histological examination of liver puncture biopsy, and the outcomes of patients were reviewed. Results Hepatomegaly, liver dysfunction, abdominal effusion and map-like density changes in liver CT scan were observed in 35 patients. Liver biopsy wag performed in 17 patients. In histopathological examination, the swelling and point-like necrosis of liver cells, expansion and congestion of sinus, endothelial swelling, wall thickening, incomplete lumen occlusion of small liver vascular were observed. Map-like density changes in liver CT scan were found in all 17 patients who were diagnosed by histological examination. Fifteen patients presented small amount of ascites within 4 weeks of onset, 13 of whom recovered or improved after treated with low-molecular weight heparin and albumin; while among the remaining 20 patients. only half of them were benefited from the same treatments. Conclusion HVOD can be diagnosed by liver CT scan instead of histological examination; treatment of patients in early stage may improve the outcome.
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Objective To investigate the expression of Toll-like receptor 2 and 4 (TLR2 and TLR4) in peripheral blood mononuclear cells (PBMCs) and the serum inflammatory cytokine levels in patients with liver failure. Methods The expressions of TLR2 mRNA and TLR4 mRNA in PBMCs were detected by RT-PCR in 20 healthy controls, 20 chronic hepatitis B (CHB) patients, 18 liver failure patients in early stage and 14 in intermediate-end stage. The serum contents of endotoxin, TNF-α and IL-6 were detected by ELISA. Results Compared with the healthy controls, the expression of TLR2 mRNA and TLR4 mRNA, and the contents of endotoxin, TNF-α and IL-6 increased in CHB patients and liver failure patients ( both early stage and intermediate-end stage) ( F = 32.997, 37.476, 23. 951,57. 265 and 38. 403, P < 0.01 ). TLR2 mRNA expression in liver failure patients in intermediate-end stage was higher than that in the early stage, but that for TLR4 mRNA was lower than that in early stage. The expressions of TLR2 mRNA and TLR4 mRNA in PBMCs were significantly correlated with the contents of TNF-α and IL-6 ( r = 0. 917 and 0. 788, P < 0. 01 ). Conclusion The inflammation reaction mediated by TLR2 and TLR4 might participate in the pathogenesis of liver failure.