Objective To compare the influence degree of laparoscopic operation and open operation on peripheral blood T lymphocyte subsets and Th1Th2 in patients with severe acute pancreatitis.Methods 54 patients who underwent surgical treatment in our hospital from February 2013 to December 2014 were selected as the subjects, 27 patients with severe acute pancreatitis who were treated with open operation as group A , 27 cases who were treated with laparoscopic operation at the same time were selected as group B , and then the peripheral blood T lymphocyte subsets and serum Th1Th2 indexes of two groups before the operation and at first,third and seventh day after the operation were respectively detected, then the detection results of two groups were compared.Results The peripheral blood T lymphocyte subsets and serum Th1Th2 indexes of group B at first, third and seventh day after the operation were all significantly better than those of group A , the difference was statistically significant ( P<0.05 ) , and the detection results of group A and group B at third day after the operation were worse than those at other time , the difference was statistically significant (P<0.05).Conclusion The influence of laparoscopic operation and open operation on T lymphocyte subsets and Th1Th2 in peripheral blood of patients with severe acute pancreatitis, the adverse effects of laparoscopic operation on the above indicators were relatively small.

Objective To prepare human anti-postsynaptic neurotoxin monoclonal antibody from phage antibody library using recombined postsynaptic short-chain neurotoxins ofLapemis curtus.Methods The three postsynaptic neurotoxins were expressed inEscherichia coli and phage antibodies against neurotoxins were screened. The obtained scFvs were further constructed to full antibodies. The antigen binding ability, biochemical and pharmacokinetic characteristics of the depurated antibodies were evaluated, and the anti-toxin effects of the antibody drugs were verifiedin vivo.Results Two positive scFvs with specific binding ability to all the three neurotoxins were obtained after 4 rounds of panning, and then the full antibodies were generated, expressed and purified. Antibody binding specificity was further confirmed. Pharmacokinetics of these two antibodies, SM-SD-911 and SM-SD-861 were similar to a conventional IgG molecule. SM-SD-911 and SM-SD-861 also showed strong antitoxin effectin vivo. Conclusionfull human anti-postsynaptic neurotoxin antibody has been successfully obtained by using recombinant neurotoxin technology and a large phage antibody library which may achieve clinical efficacy in navy medical applications.

Objective To evaluate urinary β2-microglobulin (β2-MG) and retinoid binging protein (RBP) in monitoring of early renal impairment in chronic hepatitis B (CHB) patients with long-term adefovir dipivoxil (ADV) treatment. Methods Three hundred and fifty five with CHB admitted in Shaoxing Municipal Hospital from June 2009 to June 2011 were enrolled in the study, among whom 180 cases study group) were treated with ADV monotherapy (n=100) or ADV + lamivudine (LAM) combination therapy (n=80); and 175 cases (control group) were treated with entecavir (ETV). Serum creatinine, urinary β2-MG, RBP and creatinine were measured and glomerular tration rate (eGFR) was estimated regularly during 5-year follow up. Kaplan-Meier method was used to calculate the cumulative incidence of changes in urinary β2-MG and RBP. Results Five-year follow-up results showed that in study group 2, 6, 10, 14 and 24 cases developed urinary β2-MG abnormality in year 1, 2, 3, 4 and 5 of treatment, respectively; and 2, 7, 11, 16 and 20 cases developed urinary RBP abnormality in year 1, 2, 3, 4 and 5 of treatment, respectively; eGFR decreased 20%-30% from baseline in 20 cases, 30%-50% in 13 cases and >50% in 2 cases. The decrease of eGFR ≥30% in 5 years was significantly correlated with urinary RBP and β2-GM abnormality. However, both serum creatinine and eGFR remained stable during the 5 years of follow-up in control group; only 2 cases developed urinary β2-MG abnormality and 3 cases developed urinary RBP abnormality. Conclusions Urinary RBP and β2-MG are sensitive biomarkers of early renal injury during long-term ADV treatment in CHB patients, and ADV should not be used as first-line treatment for CHB.

Objective To compare the impact of Telbivudine (LDT) and Entecavir (ETV) administration on estimates of glomerular filtration rate for anti-viral therapy in patients with hepatitis B virus (HBV)-related compensated cirrhosis by an open, prospective randomized controlled study.Methods Patients with HBV-related compensated cirrhosis at clinic or hospitalized in Shaoxing Municipal Hospital from January 2012 to June 2013 were included.A total of 170 patients were randomly divided into LDT (600 mg/d) or ETV (0.5 mg/d) groups at a ratio of 1∶1 according to the random number table method.All patients were treated for more than 36 months.The LDT group was optimized according to the roadmap.Patients with poor response or resistance in both treatment group were added with Adefovir dipivoxil (ADV) 10 mg/d for optimal treatment.The clinical outcome, creatinine (CR), estimated glomerular filtration rate (eGFR) of patients before and after 36 months of treatment were compared between two groups.All categorical data were analyzed using chi-square test and data accorded with normal distribution were compared by t test.Results After 36 months of treatment, the virological and biochemical responses in LDT group and ETV group were similar.The mean CR levels at month 24 and 36 in LDT group were (74.25±22.98) μmol/L and (70.72±24.75) μmol/L, respectively, which were both lower than baseline level ([83.09±17.68] μmol/L, t=2.811 and 3.145, respectively, both P<0.01).The mean CR levels at month 36 between two groups were statistically different (t=3.431, P=0.001).The mean eGFR levels at month 12, 24 and 36 in LDT group were all significantly lower than that at baseline (t=3.976,8.297 and 10.629, respectively, all P<0.01).The mean eGFR levels at month 24 and 36 between two groups were statistically different (t=9.684 and 15.019, respectively, both P<0.01).A total of 64 patients including 34 in LDT group and 30 in ETV group had mild nephritic injury at baseline.The mean eGFR in patients with mild nephritic injury at baseline in LDT group at month 12, 24 and 36 were significantly different compared to baseline (t=6.098,10.191 and 14.378, respectively, all P<0.01).The mean eGFR level at month 36 in ETV group had statistical difference compared to baseline (t=2.058, P<0.05).The mean eGFR levels at months 12, 24 and 36 were all statistical different between two groups (all P<0.01).The mean eGFR levels at month 24 and 36 in the optimized group were superior to ETV group (P<0.01).Conclusions In patients with HBV-related compensated cirrhosis, LDT and ETV treatment have similar clinical efficacy.LDT is more effective in protecting nephritic function than ETV.

Objective:To investigate the clinical efficacy and safety of fruquintinib in elderly patients with advanced metastatic colorectal cancer who failed chemotherapy.Methods:Ninety-nine elderly patients with advanced metastatic colorectal cancer who failed chemotherapy in No. 904 Hospital of Joint Logistics Support Force from September 2018 to July 2020 were selected. All patients were given furquintinib capsules, 1 time/d, 5 mg/time, and 28 days was 1 cycle. All patients were treated continuously for 2 cycles and the effect was observed. The patient's recent anti-tumor efficacy was counted. The serum levels of carbohydrate antigen 125 (CA125), carcinoembryonic antigen (CEA) and carbohydrate antigen 199 (CA199) in patients before and after treatment were compared. The safety of the medication during the patient's treatment was recorded, and the Kaplan-Meier method was used for survival analysis.Results:A total of 99 elderly patients with advanced metastatic colorectal cancer who failed chemotherapy were treated for 2 cycles, with an objective response rate (ORR) of 22.22% (22/99) and a clinical control rate (CCR) of 75.76% (75/99). The serum levels of CA125, CA199 and CEA after treatment were lower than those before treatment (all P<0.05). The drug adverse reactions in 99 patients during the treatment were mostly grade Ⅰ-Ⅱ, and grade Ⅲ-Ⅳ were rare. The most common gradeⅠ-Ⅱ adverse reactions were hypertension (45.45%, 45/99), hand-foot syndrome (40.40%, 40/99), and elevated aspartate transferase (36.36%, 36/99). Followed up for 12 months, 5 cases were lost to follow-up, the follow-up rate was 94.95%, the median progression-free survival time of the remaining 94 patients was 5.62 months (95% CI 3.57-8.75 months), and the median overall survival time was 8.41 months (95% CI 4.85-11.14 months). Conclusions:Fruquintinib has good efficacy in the treatment of elderly patients with advanced metastatic colorectal cancer who failed chemotherapy. It can reduce the levels of tumor markers, the survival status of patients is good, and the adverse reactions are controllable.

M2 protein of influenza A virus is encoded by a spliced mRNA derived from RNA segment 7 and plays an important role in influenza virus replication. It is also a target molecule of anti-virus drugs. We extracted the viral genome RNAs from MDCK cells infected with swine influenza A virus (SIV) H3N2 subtype and amplified the SIV M2 gene by reverse transcriptase-polymerase chain reaction using the isloated viral genome RNAs as template. The amplified cDNA was cloned into a prokaryotic expression vector pET-28a(+) (designated pET-28a(+)-M2) and a eukaryotic expression vector p3xFLAG-CMV-7.1 (designated p3xFLAG-CMV-7.1-M2), respectively. The resulted constructs were confirmed by restriction enzyme digestion and DNA sequencing analysis. We then transformed the plasmid pET-28a(+)-M2 into Escherichia coli BL21 (DE3) strain and expressed it by adding 1 mmol/L of IPTG (isopropyl-beta-D-thiogalactopyranoside). The recombinant M2 protein was purified from the induced bacterial cells using Ni(2+) affinity chromatography. Wistar rats were immunized with the purified M2 protein for producing polyclonal antibodies specific for it. Western blotting analysis and immunofluorescence analysis showed that the produced antibodies were capable of reacting with M2 protein expressed in p3xFLAG-CMV-7.1-M2-transfected cells as well as that synthesized in SIV-infected cells. We also transfected plasmid p3xFLAG-CMV-7.1-M2 into Vero cells and analyzed its subcellular localization by immunofluorescence. The M2 protein expressed in the Vero cells was 20 kDa in size and dominantly localized in the cytoplasm, showing a similar distribution to that in SIV-infected cells. Western blotting analysis of SIV-infected cells suggested that M2 was a late phase protein, which was detectable 12 h post-infection, later than NS1, NP and M1 proteins. It would be a potential molecular indicator of late phases replication of virus. Our results would be useful for studying the biological function of M2 protein in SIV replication.
Animals ; Antibodies, Monoclonal ; biosynthesis ; Cercopithecus aethiops ; Cloning, Molecular ; Escherichia coli ; genetics ; metabolism ; Influenza A Virus, H3N2 Subtype ; genetics ; RNA ; biosynthesis ; genetics ; Rats ; Rats, Wistar ; Recombinant Proteins ; biosynthesis ; genetics ; immunology ; Swine ; Transfection ; Vero Cells ; Viral Matrix Proteins ; biosynthesis ; genetics ; Virus Replication ; genetics

Influenza A virus matrix protein (M1) is encoded by a spliced mRNA derived from RNA segment 7 and plays an important role in the virus life cycle. In the present study, we extracted the viral genome RNAs from allantoic fluid of 9-day-old embryonated chicken eggs infected with swine influenza A virus (SIV) H3N2 subtype and amplified the SIV M1 gene by reverse transcriptase-polymerase chain reaction using the isloated viral genome RNAs as template. The amplified cDNA was cloned into an expression vector pET-28a (+) (designated pET-28a-M1) and confirmed by DNA sequencing analysis. We then transformed the plasmid pET-28a-M1 into Escherichia coli BL21 strain for heterologous expression. The expression of M1 was induced by 1mM isopropyl-beta-D-thiogalactopyranoside. SDS-PAGE analysis of the induced bacterial cells revealed that the recombinant M1 protein was expressed in high yield level. Next, we purified the expressed recombinant M1 using Ni2+ affinity chromatography and immunized Wistar rat with the purified M1 protein for producing polyclonal antibodies specific for M1. Western blotting analysis showed that the produced antibodies were capable of reacting with M1 protein expressed in Escherichia coli as well as that synthesized in SIV-infected cells. We further cloned the amplified M1 cDNA into a eukaryotic expression plasmid p3xFLAG-CMV-7.1 to construct the recombinant plasmid p3xFLAG-CMV-M1 for expressing M1 in eukaryotic cells. Western blotting analysis revealed that the M1 protein was expressed in p3xFLAG-CMV-M1-transfected Vero cells and recognized by the produced anti-M1 antibodies. Using the produced anti-M1 antibodies, we analyzed the kinetics of M1 protein in the virus-infected cells during influenza virus infection and estimated the possibility of M1 as an indicator of influenza virus replication. The recombinant M1 protein, anti-M1 antibodies and recombinant expression plasmids would provide useful tools for studies of biological function of M1 protein and the basis of SIV replication.
Animals ; Antibodies, Monoclonal ; biosynthesis ; Chick Embryo ; Cloning, Molecular ; Escherichia coli ; genetics ; metabolism ; Influenza A Virus, H3N2 Subtype ; genetics ; physiology ; Rats ; Rats, Wistar ; Recombinant Proteins ; genetics ; immunology ; metabolism ; Swine ; Viral Matrix Proteins ; genetics ; immunology ; metabolism ; Virus Replication ; genetics

Objective:To investigate the clinical characteristics, gene mutation, treatment and prognosis of familial aggregation myelodysplastic syndromes/acute myeloid leukemia (MDS/AML).Methods:The 3 familial aggregation MDS/AML admitted to Shanghai Yangpu District Hospital from August 2012 to March 2019 were collected. The bone marrow examination, gene mutation detection, therapeutic effect and prognosis of the patients were retrospectively analyzed, and the relevant literature was reviewed.Results:In pedigree 1, the survival time of 2 AML patients was 8 months and 1 month, respectively. In pedigree 2, the transformation time of 2 patients diagnosed MDS to AML/high-risk MDS was 4 and 3 months, the survival time was 5 and 8 months, respectively. TP53 and RUNX1 mutations were detected in the older brother. In pedigree 3, the survival time of the AML patient was 13 months, and the MDS patient was stable.Conclusions:Familial aggregation MDS/AML has rapid progression and short survival time, and its diagnosis needs to be combined with family history, cytogenetics, and molecular biology.

Objective:To analyze the incidence of liver function injury in patients infected with 2019-nCoV omicron variant and its influencing factors.Methods:The clinical data and laboratory findings of 897 COVID-19 patients infected with omicron variant in Zhejiang province from February 23 to July 14, 2022 were retrospectively analyzed. Patients were divide into liver function injury group ( n=243) and non-liver function injury group ( n=654) based on liver function indicators. The clinical characteristics and laboratory tests were compared between the two groups, and influencing factors of liver function injury were analyzed. SPSS 26.0 statistical software was used for data analysis. Results:The incidence of liver injury in this series was 27.09% (243/897). The median age of patients in liver injury group was older, the body mass index (BMI) was higher( Z=-6.237 and -2.166, both P<0.05), the proportions of patients with hypertension and diabetes, and with severe clinical classification were higher ( χ2=17.087, 27.509 and 12.945, all P<0.01) ; the proportion of vaccinated patients was lower ( χ2=17.766, P<0.01) than those in non-liver injury group. The levels of platelet, hemoglobin, albumin and potassium in liver injury group were lower than those in non-liver injury group ( Z=-4.631, -2.368, -10.593 and -2.141, all P<0.05), while serum ALT, AST, γ-GT, urea nitrogen, glucose and hs-CRP levels were higher than those in the non-liver injury group ( Z=-7.451, -8.663, -4.410, -3.824, -3.278 and -3.884, all P<0.01). Multivariate Logistic regression analysis showed that age ( OR=2.580, 95% CI 1.429-4.657, P=0.002), history of diabetes ( OR=3.650, 95% CI 1.698-7.849, P=0.001), and decreased hemoglobin ( OR=1.993, 95% CI 1.066-3.726, P=0.031) and increased hs-CRP ( OR=1.797, 95% CI 1.283-2.517, P=0.001) were risk factors associated with liver function injury, while vaccination ( OR=0.499, 95% CI 0.312-0.798, P=0.004) was the protective factor for liver function. Conclusion:Liver function injury is frequently observed in COVID-19 patients infected with omicron variant, which is linked to age, underlying disease, and elevated inflammatory markers; while vaccination can lower the risk of liver injury in infected patients.

Objective To analyze the changes in follicular helper T (Tfh) cells during HIV-1 in-fection, to investigate the influences of Tfh cells and Tfh-related molecules on HIV-1 progression and to pro-vide references for further research on using Tfh cells in highly active antiretroviral therapy ( HAART) and vaccines. Methods This study enrolled 33 patients with HIV-1 infection, including 11 long-term nonpro-gressors (LTNP), 10 rapid progressors (RP) and 12 typical progressors (TP), and 11 healthy subjects (normal controls, NC). Peripheral blood mononuclear cells were isolated from each subject. Multicolor flow cytometry was performed to detect CD4+CD45RA-CXCR5+Tfh and CD4+CD45RA-CXCR3-CXCR5+PD-1+Tfh subsets and the levels of inducible costimulatory molecule (ICOS), IFN-γ and IL-21. Moreover, the levels of IL-10 and the percentages of CD19+B cells in plasma samples of each group were also analyzed. Relationships among Tfh, CD4 and B cells were analyzed. Results The percentages of both Tfh subsets were higher in patients with HIV-1 infection than in NC. Compared with NC, LTNP had the highest percent-age of CD4+CD45RA-CXCR3-CXCR5+PD-1+Tfh cells (P<0. 05). Expression of Tfh-related molecules ICOS, IFN-γ and IL-21 were enhanced significantly upon Staphylococcus enterotoxin B ( SEB) stimulation, ICOS+Tfh cells were negatively related with HIV-1 progression, but had a positive correlation with CD19+B cells (r=-0. 49, P<0. 01; r=0. 60, P<0. 05). IL-10 level in plasma increased significantly in patients withHIV-1 infection , especially in TP and RP ( TP vs NC : P＜0. 01 ; RP vs NC : P<0. 05 ) . Conclusion HIV-1 patients and NC had significant differences in the expression of Tfh cells and Tfh-related molecules in peripheral blood. ICOS+Tfh cells were closely related to the progression of HIV-1 infection and the function of B cells.