Construction of SARS-CoV-2 receptor binding domain molecular probe and its application in the isolation of neutralizing antibodies / 中国热带医学

@#Abstract: Objective　To construct SARS-CoV-2 receptor binding domain molecular probe for monoclonal memory B cell sorting and obtain RBD specific neutralizing antibodies from peripheral blood mononuclear cells (PBMCs) of COVID-19 convalescents by single-cell sorting. Methods　The SARS-CoV-2 RBD sequence was downloaded from GenBank, and the Avi tag and 6-histidine tags were added at the C-terminal. After codon optimization, it was chemically synthesized, cloned into the pDRVI1.0 vector, expressed after transfection of 293F cells, and biotinylated consequently. RBD-specific B cells were sorted out with this probe1 from the PBMCs of convalescents recovered from COVID-19. After B cells were lysed, the variable regions of heavy chain and light chain were amplified, cloned into the antibody expression vector, and transfected into 293F cells to express the antibody. Then the antibody was purified from the supernatant using protein A column and SARS-CoV-2 pseudovirus was used to test their neutralizing activity. Results　RBD-Avi probe was produced and successfully biotinylated sequentially with an efficiency of 30%-50%. Western blot analysis revealed that the biotinylated probe was recognized by the antibodies purified from COVID-19 convalescent plasma. Using this probe, 7 and 16 RBD-specific memory B cells were successfully isolated from the PBMCs of two convalescent individuals, accounting for 0.24% and 0.17% of the total cell population, respectively. After amplifying the variable regions of antibody heavy and light chains from the lysed B cells, 7 and 12 pairs of antibody heavy-light chains were obtained. A total of 16 antibodies were expressed in the convalescent individuals, and most of the purified antibodies showed neutralizing activity against the pseudovirus, with IC50 values of 6 antibodies below 1 μg/mL. The IC50 values of XJ-A9 and SCF-F1 against the wild-type pseudovirus were 0.07 μg/mL and 0.35 μg/mL, respectively. Conclusion　The SARS-CoV-2 RBD molecular probe constructed in this study has good antigenicity, and the isolated antibodies present neutralizing activity against wild-type SARS-CoV-2 pseudovirus.

HIV-1 Subtype Diversity and Factors Affecting Drug Resistance among Patients with Virologic Failure in Antiretroviral Therapy in Hainan Province, China, 2014-2020 / 生物医学与环境科学（英文）

OBJECTIVE@#This study aimed to determine the HIV-1 subtype distribution and HIV drug resistance (HIVDR) in patients with ART failure from 2014 to 2020 in Hainan, China.@*METHODS@#A 7-year cross-sectional study was conducted among HIV/AIDS patients with ART failure in Hainan. We used online subtyping tools and the maximum likelihood phylogenetic tree to confirm the HIV subtypes with pol sequences. Drug resistance mutations (DRMs) were analyzed using the Stanford University HIV Drug Resistance Database.@*RESULTS@#A total of 307 HIV-infected patients with ART failure were included, and 241 available pol sequences were obtained. Among 241 patients, CRF01_AE accounted for 68.88%, followed by CRF07_BC (17.00%) and eight other subtypes (14.12%). The overall prevalence of HIVDR was 61.41%, and the HIVDR against non-nucleoside reverse transcriptase inhibitors (NNRTIs), nucleotide reverse transcriptase inhibitors (NRTIs), and protease inhibitors (PIs) were 59.75%, 45.64%, and 2.49%, respectively. Unemployed patients, hypoimmunity or opportunistic infections in individuals, and samples from 2017 to 2020 increased the odd ratios of HIVDR. Also, HIVDR was less likely to affect female patients. The common DRMs to NNRTIs were K103N (21.99%) and Y181C (20.33%), and M184V (28.21%) and K65R (19.09%) were the main DRMs against NRTIs.@*CONCLUSION@#The present study highlights the HIV-1 subtype diversity in Hainan and the importance of HIVDR surveillance over a long period.

Blockage of TGF-β1-induced epithelial-to- mesenchymal transition by oxymatrine prevents renal interstitial fibrosis

Abstract Most chronic kidney disease inevitably progress to renal fibrosis. Tubular epithelial- to-mesenchymal transition (EMT) is recognized to play major roles in renal fibrosis. Oxymatrine (OM) is a major alkaloid component found in a Chinese herb Sophora roots and has many effects. The aim is to investigate the effect of OM on renal tubular EMT and elucidate its mechanism. Mice underwent unilateral ureteral obstruction (UUO) followed by intraperitoneal injection of OM (120 mg/kg) or control vehicle. Human kidney proximal tubular cell line (HK-2) was used and EMT was induced with 5 ng/mL of transforming growth factor-β1 (TGF-β1). In vivo, renal tubulointerstitial fibrosis was induced and E-cadherin was down-regulated, while the expressions of fibronectin (FN), α-smooth muscle actin (α-SMA), TGF-β1 and its type I receptor (TGF-βRI) were up-regulated in UUO mice. In contrast, OM significantly ameliorated renal fibrotic lesions and attenuated the expressions of FN, α-SMA, TGF-β1 and TGF-βRI, but increased E-cadherin in the obstructed kidneys. In vitro, OM abolished TGF-β1-mediated E-cadherin suppression and FN, α-SMA and TGF-βRI induction in HK-2 cells in a dose-dependent manner. These observations strongly suggest that the renal protective effects of OM could be mediated by prevention of EMT and manifested as suppression of TGF-β1 and TGF-βRI expressions.

Clinical Characteristics, Prognostic Factors and Misdiagnosis of Bone Marrow Necrosis Patients / 中国实验血液学杂志

OBJECTIVE@#To analyze the clinical characteristics, diagnosis and prognostic factors of bone marrow necrosis (BMN) patients, aim to avoid misdiagnosis, missed diagnosis or delayed treatment.@*METHODS@#The clinical data of 51 BMN patients treated in the Affiliated Hospital of Xuzhou Medical University from January 2010 to December 2017 were retrospectively analyzed. The types of primary disease, etiology, clinical manifestations, laboratory tests, radiological findings, treatment outcomes and prognostic factors were summrized, and the reasons for misdiagnosis were analyzed.@*RESULTS@#Among 51 BMN patients, the hematologic tumor was detected out in 32 patients; solid tumors caused- BMN was detected out in 14 patients, benign lesions for 5 patients. The time of interval from the appearance of symptoms to the confirmation of BMN was 7 days to 6 months, with a median of 35 days. Misdiagnosis and missed diagnosis occurred in 25.5% of the BMN patients. Anemia was found in all of the 51 BMN patients, fever accounted for 58.8%, systemic bone pain for 52.9%, bleeding for 29.4%, lymphadenectasis for 37.3%, and hepatosplenomegaly for 19.6%. Leukoerythroblastic anemia accounted for 84.3%, bicytopenia for 51.0%, pancytopenia for 25.5%, and monocytopenia for 23.5%. The serologic test revealed no specific results. The first bone marrow aspiration were 38 patients and multi-site puncture were 7 patients. The diagnostic coincidence rate of bone marrow smear was 88.2%. Among 51 BMN patients, 41 patients received bone marrow biopsy, and the diagnostic coincidence rate of bone marrow biopsy was 75.6%. The abnormal signals were found in multiple vertebral bodies by spinal/pelvic MRI scan in 13 BMN patients; PET-CT scan revealed a diffuse pattern of low FDG uptake in the bone marrow in 16 patients, with a local increase in FDG uptake accompanied by bone marrow involvement. For 46 patients with BMN combined with malignancies, among which 35 patients died (76.1%) and the median survival time was 25 days. Among the 32 patients with hematologic tumors, early death occurred in 12 patients, BMN disappeared in 11 out of 20 patients received active chemotherapy for the primary disease, 9 patients died within 1 week to 3 months. Fourteen patients combined with bone marrow metastatic carcinoma died within 2 weeks to 3 months. Focal necrosis disappeared in 4 out of 5 BMN patients secondary to non-malignant diseases after symptomatic supportive treatment and still alived. Multiple logistic regression was performed to analyze factors affecting the prognosis of BMN patients, the result showed that the prognosis of BMN was closely related to the factors of primary disease (benign and malignant). The reasons for misdiagnosis and missed diagnosis were as follows: hidden onset of the primary disease, nonspecific symptoms, insufficient understanding and alertness of the physicians regarding the primary clinical characteristics and hematological abnormalities, and failure to receive multiple sites bone marrow punctures or bone marrow biopsies.@*CONCLUSION@#BMN usually occurs concomitantly to hematologic tumors and bone marrow metastases from solid tumors. Its prognosis is closely related to the nature and severity of the primary disease and its own severity. In the clinic, BMN should be suspected in patients with severe bone pain, fever, hepatosplenomegaly, hemocytopenia, lymphadenectasis and leukoerythroblastic anemia. Bone marrow puncture at multiple positions and bone marrow biopsy can compensate for each other in the diagnosis of BMN. The combined use of the two methods can improve the diagnostic coincidence rate of BMN, and the positive rate of the etiological diagnosis of BMN.

Research progress on factors affecting the establishment and development of oral microbiota in early life / 口腔疾病防治

@#The oral microbiota has been proven to play an essential role in local and systemic health. Hence, it is essential to understand how the oral microbiota and further microbial ecosystems are established and the associated development process to understand the physiologic and pathologic events related to the oral microbiota and management of the whole lifecycle oral and systemic health. The present review summarizes recent studies of the establishment and development of oral microbial communities in early life with related factors and briefly describes their prospects. By investigating the characteristics and development of the oral microbiota, current studies have shown that health status during pregnancy, mode of delivery, diet, environment, antibiotics and genetic factors are related to oral microbiota development, which is helpful in clinical practice. Nevertheless, current studies are mostly cross-sectional. Prospective cohort studies with larger sample sizes using metagenomics or metatranscriptomics and studies on molecular mechanisms are needed to further elucidate the influences and mechanisms of the related factors on the establishment and development of microbiota in early life and their impact on local or systemic health.

Material Basis of Xuanfei Huazhuo Prescription in Treatment of COVID-19 by Multi-target Molecular Docking / 中国实验方剂学杂志

Objective:Structure-based angiotension converting enzyme 2 (ACE2) and interleukin-6R (IL-6R) were taken as the target proteins to in the investigation of the material basis of Xuanfei Huazhuo prescription in the treatment of coronavirus disease-2019 (COVID-19) by molecular docking. Method:The compounds in Xuanfei Huazhuo prescription were retrieved through TCMSP. Structure-based ACE2 and IL-6R were taken as the target proteins to screen out the compounds with a better activity by molecular docking, and analyze structural properties of these compounds. Furthermore, the potential molecular mechanism of Xuanfei Huazhuo prescription in the treatment of COVID-19 was analyzed by target reverse prediction. Result:There were 312 potentially active compounds in Xuanfei Huazhuo prescription, including 75 highly active compounds and 15 highly active compounds for ACE2. There were 100 eligible active compounds and 3 highly active compounds for IL-6R, most of which belong to flavonoids. The herb-component-target network included 10 herbs, 126 compounds and 130 targets. String analysis showed that PIK3R1, SRC, AKT1, AR and EGFR might be the key targets of Xuanfei Huazhuo prescription. Conclusion:Based on the virtual screening of multi-target molecular docking, the anti-virus and anti-inflammatory material basis of Xuanfei Huazhuo prescription was preliminarily obtained. At the same time, based on the reverse prediction and analysis, potential targets and molecular mechanism of the recipe in the treatment of COVID-19 were explored, so as to provide clues for the multi-angle mining of Xuanfei Huazhuo prescription and its relevant prescriptions and the modernization development of monomer components.

Possile Effect of Small Molecule in Hedysari Radix in Antagonizing Tumor Necrosis Factor Receptor 1 Based on Molecular Docking / 中国实验方剂学杂志

Objective: To study the effect of small molecule compounds of Hedysari Radix in ntagonizing tumor necrosis factor receptor type 1 (TNFR1) based on molecular docking. Method: The structure of small molecular compound of Hedysari Radix was downloaded from the chemical composition compound library of traditional Chinese medicine, and then optimized to obtain the composition compound library of Hedysari Radix. The three-dimensional structure of the inflammatory target TNFR1 (PDB ID:1TNR) was identified. After hydrotreating and anhydrating, the binding pocket residues were identified according to the literature. According to the defined target structure and binding pocket, the flexible molecular docking was conducted between the composition compound library and the target, and the score (Glide Score) was obtained. Based on the results of molecular docking, the first nine small molecular compounds of Glide Score were selected as candidate components. On this basis, the drug-likeness was analyzed, which involved small molecular compounds that meet the number of hydrogen-bonded receptors, the number of hydrogen-bonded donors, the formula weight, the number of rotatable key and the numerical range of lipo-hydro partition coefficient. Finally, the binding mode was analyzed according to pharmacokinetic parameters and complex structure of composition-target docking. Result: The residue set in the TNFR1 drug-binding pocket were identified as glutamic acid109 (Glu109), lysine 35(Lys35), alanine62 (Ala62), serine 74 (Ser74), lysine75 (Lys75), cysteine76 (Cys76), argnine 77(Arg77), glutamine82 (Gln82), threonine89 (Thr89), asparticacid91 (Asp91), argnine92 (Arg92), aspartic acid93 (Asp93), threonine 94(Thr94), valine95 (Val95), cysteine 96(Cys96), argnine104 (Arg104), tyrosine106 (Tyr106), asparagine110 (Asn110), leucine111 (Leu111), phenylalanine112(Phe112), glutamic acid 131(Glu131) and lysine132 (Lys132). Totally 43 small molecular compounds of Hedysari Radix were obtained. Five small molecular compounds, namely hedysari radix, quercetin, isoliquiritin, naringenin, calycosin and liquiritigenin, were screened by comprehensive factors, like docking scoring. Conclusion: Quercetin, isoliquiritin, naringenin, calycosin and liquiritigenin are the effective anti-inflammatory substances of Hedysari Radix, with a great possibility of becoming TNFR1 antagonists.

Quality Relevance of Standard Decoction and Different Drying Methods Products for Gentianae Macrophyllae Radix Granules Based on HPLC Fingerprint / 中国实验方剂学杂志

Objective:To establish the fingerprints of standard decoction of Gentianae Macrophyllae Radix-dried products by different methods,and to evaluate the quality correlation. Method:HPLC,InertSustain C18 chromatographic column(4.6 mm×250 mm,5 μm),Gradient elution was performed for the mobile phase of acetonitrile-phospho,detection wavelength at 240 nm and flow rate of 0.8 mL·min-1,and column temperature was 40℃. The quality correlation analysis of different methods for different kinds of Gentianae Macrophyllae Radix standard decoction was carried out from the aspects of chemical composition consistency,common chemical composition consistency,main chemical composition content and transfer rate. Result:The control fingerprint of Gentianae Macrophyllae Radix standard decoction was established. According to the peak matching data,there were 10 common peaks in the fingerprint of 15 batches of Gentianae Macrophyllae Radix standard decoction.Among the 10 common peaks,5 chemical constituents of loganic acid,6'-O-β-D-glucosyl gentiopicroside,swertiamarin,gentiopicroside and swertia glycosides were identified. The results of quality correlation analysis showed that the three different drying methods were consistent with the chemical composition and quantity of Gentianae Macrophyllae Radix standard decoction. But in terms of the content consistency of common chemical components and the transfer rate of main chemical components,the quality correlation between the products obtained from vacuum drying and the standard decoction was lower than that obtained from spray drying and freeze-drying. Conclusion:The fingerprint of different method of Gentianae Macrophyllae Radix standard decoction was established. Through the analysis of the mass correlation of chemical composition consistency,common chemical composition content consistency,main chemical composition content and transfer rate,the mass correlation between them was comprehensively reflected. It is suggested that spray drying or freeze drying should be used for the key drying process of Gentianae Macrophyllae Radix granules. This study provides a reference for the preparation process and quality control of Gentianae Macrophyllae Radix granules.

The Application of Hierarchical Chain Nursing in Pediatric Pneumonia Nursing / 昆明医科大学学报

Objective To explore the effect of stratified chain nursing model on the efficacy, symptom improvement and nursing satisfaction in pediatric pneumonia nursing. Me thods 58 cases of pediatric pneumonia in our hospital from March 2015 to March 2016 were selected as the control group. 58 cases of pediatric pneumonia in our hospital from April to April 2017 were selected as observation group. After 2 weeks of care, the two groups were compared in the curative effect, the symptoms improved and the nursing satisfaction. Re s ults The total effective rate of the two groups was significantly higher than that of the control group (P＜0.05). The disappearance time of fever, the time of disappearance of cough, the time of disappearance of lung rales and the time of hospitalization were shorter than those of the control group, the difference was statistically significant (P ＜0.05). The observation group had higher evaluation scores in ward environment, nursing technology, sense of responsibility, health education, psychological care, and service attitude than the control group, the difference was statistically significant (P ＜0.05). Conclus ion Hierarchical chain nursing model is effective in the treatment of pediatric pneumonia, which can effectively improve the curative effect and nursing satisfaction, which is beneficial to the improvement of symptoms.

Establishment of orthotopic implantation model of human U87-MG brain glioma cell line in nude mice / 中国药理学通报

Aim To establish human U87-MG glioma model in nude mice brain and to observe the characteristics of the tumor growth. Methods Human U87-MG glioma cells were cultured in vitro. 5 μL of cell suspension containing 3.0 ×1010·L-1, 4.0×1010·L-1and 5.0×1010·L-1respectively was inocula-ted into the right caudate nucleus of 18 male nude mice brain un-der the guidance of stereotaxic apparatus, separately, whereas another 6 nude mice as the control group, were inoculated into the same volume of Hanks solution. The moving and survival state of rats with gliomas were observed. The examinations of the tumors formation, volumes, metastasis and histopathology were performed and the obtained brain samples were stained with HE and immunohistochemistry. Results All the tested rats of dif-ferent inoculation doses developed brain tumors without extracra-nial metastasis. The mean survival time of three groups was (46.50 ± 3.27) d,(38.50 ± 3.28) d and (30.67 ± 3.51) d,respectively. The tumors showed the similar morphological fea-tures and immunophenotype to human glioma. There was positive expression of GFAP and S-100 in the tumors. Conclusions The orthotopic implantation model of human U87-MG glioma, by in-oculating quantitative U87-MG cells stereotaxically into the brains of the nude mice, is successfully established with 100 yield of intracranial tumor and no extracranial growth extension. It resembles the histopathological and morphological features of human glioma,which can be used as a reliable animal model for the study of the tumorigenesis, pathogenesis, biological charac-teristics and therapy of glioma.

Effects of Astragalus Lily Granules on enhanced hypoxia tolerance in mice exposed to sodium nitrite poisoning / 西安交通大学学报(医学版)

Objective To study the effect of Astragali Lilium Granules on the survival time,CAT,ATP,and pathological changes in mice exposed to sodium nitrite (NaNO2)poisoning.Methods We randomly divided 60 male mice of SPF grade KM into blank group,positive control group,model group and Astragali Lilium Granules low-,medium-and high-dose groups.After 3 days of normal feeding,blank group and model group received intragastric gavage of normal saline,and positive control group was given Rhodiola rosea capsule solution.Astragali Lilium Granules groups received intragastric gavage according to the concentration (1.75,3.5 and 7 mg/kg per day) for 30 days. One hour after the last administration, all groups except the blank control group received intraperitoneal injection of NaNO2(200 mg/kg,injection amount of 0.1 mL/g).We recorded the mice's survival time,observed the pathological changes of brain,lung and heart tissues by HE staining under the microscope.The activity of CAT and the level of ATP in brain tissue were determined by colorimetric analysis.Results Compared with the blank group,the survival time in model group [(9.64±1.60)min,P＜0.05]was significantly lower;in model group,the lung tissue showed extensive pulmonary edema,red cell exudation,emphysema,and local atrophy of the lung;the interstitium of the myocardium was dilated and congested,and the local cells in the epicardium became edematous and denatured;brain cells showed necrosis,cytoplasm condensation,pyknosis,and perinuclear vacuoles.The tissues were loose and the spaces between the vessels and nerve cells widened;the activity of CAT decreased and the content of ATP decreased in lung,heart and brain tissues (P＜0.05).Compared with the model group,the survival time was significantly prolonged in Astragali Lilium Granules low-[(12.78±2.20)min]and medium-dose [(13.22±2.10)min]groups (P＜0.05).Compared with model group,the three dose groups had lessened lung tissue edema and emphysema,cardiac interstitial vascular dilatation and congestion,necrosis and nuclear pyknosis of brain nerve cells;the activity of CAT increased and the content of ATP increased in lung,heart and brain tissues (both P＜0.05).Conclusion Astragali Lilium Granules can improve the toxicity of NaNO2in mice and thus effectively protect against NaNO2-induced anoxic injury.

Establishment of Jinzepaishi Granules fingerprint and evaluation of its anti-inflammatory activity for cyclooxygenase-2 / 中草药

Objective To establish HPLC fingerprints of Jinzepaishi Granules from ten different batches and a preliminary method for its anti-inflammatory biological activity titer detection, and construct a dose-effect relationship between cyclooxygenase-2 (COX-2) and Jinzepaishi Granules for scientific evaluation and effective control of granules quality. Methods The HPLC method was used to analyze the similarity of common peaks in different batches of granules and assigned 19 common peaks to herbs. The fingerprints were further analyzed by the chemometric method of hierarchical clustering analysis and principal component analysis (PCA). The EIA method was used to determine the half-inhibitory concentration (IC50) of Jinzepaishi Granules. Results The similarity of HPLC fingerprints of all the ten batches was ≥ 0.955, which were consistent with the results of cluster analysis and PCA. Jinzepaishi Granules had a significant inhibitory effects on COX-2 with a IC50 of 10.272 mg/mL, which demonstrated a significant dose-effect relationship. Conclusion The multiple quality control system of chemical fingerprinting and biological activity detection has the characteristics of high efficiency, rapidness, accuracy, sensitivity, and good reproducibility. It can effectively evaluate the anti-inflammatory biological activity of Jinzepaishi Granules and achieve more scientific and reasonable control of its quality. It also provides a reference for the establishment of more scientific quality control for other Chinese materia medica preparations.

Clinical Study on Electronic Moxibustion in Treating Diabetic Peripheral Neuropathy / 上海针灸杂志

Objective To observe the clinical efficacy of electronic moxibustion in treating diabetic peripheral neuropathy due to liver-kidney deficiency. Method Ninety patients with diabetic peripheral neuropathy due to liver-kidney deficiency were randomized into a treatment group of 44 cases and a control group of 46 cases. The treatment group was intervened by acupoint therapy by warm electronic moxibustion, while the control group was intervened by foot bath with Chinese medication. For both groups, 10 treatment sessions were taken as a course of treatment, for 3 courses in total. Before and after the treatment, the whole blood viscosity, blood lipids, fasting blood glucose (FBG), glycosylated hemoglobin (GHb) and Michigan Neuropathy Screening Instrument (MNSI) were evaluated in the two groups. Result The whole blood viscosity indexes, blood lipids levels, FBG level and GHb level didn't show significant differences after the treatment in the two groups (P>0.05). The intra-group and inter-group comparisons of MNSI score showed significant differences after the treatment (P<0.01); there was a between-group difference in the change of MNSI score after the treatment (P<0.01). Conclusion Electronic moxibustion is an effective approach in treating diabetic peripheral neuropathy.

Protective effects of Astragalus immortal prescription against liver oxidative stress in irradiated mice / 解放军医学杂志

Objective To explore the protective effect and possible mechanism of Astragalus immortal prescription (AIP) from hepatic oxidative stress in mice irradiated by X ray.Methods Six to eight weeks old Kunming mice were randomly divided into normal control group,irradiation alone group,positive control group,and low,medium and high dose groups of AIP (10 rats in each group),physiological saline,thymus peptide and different concentration decocta of AIP were given respectively.After continuous gavage for 10 days,the whole body was irradiated with X ray at the dose of 8 Gy at a time,and then continued lavaging for 3 days,with weighing the mice and observing the diet,water intake and general condition.After killing the mice,the liver was take and the liver index was calculated;the morphological changes of the liver tissues were observed and the expressions of superoxide dismutase (SOD),malondiadehyde (MDA),glutathione peroxidase (GSH-Px) and nuclear factor erythroid 2-related factor 2 (Nrf2) in the liver tissues were detected.Results Compared with the normal control group,the weight of mice in model group decreased obviously (P＜0.01);pathological section showed that the liver cells were disordered,the central vein was congested and blocked seriously,the surrounding liver cells were denatured and necrotic;the activity of SOD and GSH-Px in liver tissues decreased,and the content of MDA increased (P＜0.05),the expression of Nrf2 protein increased significantly (P＜0.01).Compared with the radiation alone group,the weight of mice in positive control group and high dose AIP group increased significantly (P＜0.01);pathological section showed that the liver tissue structure was basically normal,the liver cells were radially arranged with the central vein as the center;the activity of SOD and GSH-Px in liver tissues increased,and the content of MDA decreased (P＜0.05),the expression of Nrf2 protein decreased significantly (P＜0.01).Conclusion AIP has protective effect from hepatic oxidative stress in mice irradiated by X ray,and the mechanism may be related to its antioxidant effects.

Efficacy of pegylated-interferon alpha-2a treatment in patients with HBeAg-positive chronic hepatitis B and partial viral response to nucleoside analogue therapy / 中华肝脏病杂志

<p><b>OBJECTIVE</b>To investigate the efficacy and related factors of pegylated-interferon alpha-2a (PEG-IFN-2a) treatment in patients with hepatitis B e antigen (HBeAg)-positive chronic hepatitis B (CHB) who achieved partial viral response with nucleoside analogue (NA) therapy.</p><p><b>METHODS</b>Patients with HBeAg-positive CHB and partial viral response to NA treatment were administered a PEG-IFN-2a therapy regimen of 180 g subcutaneous injection once weekly for a personlized duration of time. The existing NA therapy was continued in combination with the new PEG-IFN-2a treatment for 12 weeks. Measurements of serum HBV DNA load, hepatitis B surface antigen (HBsAg), hepatitis B surface antibody (anti-HBs), HBeAg and hepatitis B e antibody (anti-HBe) were taken at baseline (prior to addition of the PEG-IFN-2a therapy) and every 3 months afterwards.For determining response to treatment, primary efficacy was defined as undetectable HBsAg and seroconversion, and secondary efficacy was defined as HBsAg less than 10 IU/mL and HBeAg seroconversion.Statistical analysis was carried out using SPSS statistical software.</p><p><b>RESULTS</b>A total of 81 consecutive patients with an average of 12.0 months (range: 6.0-24.0 months) of NA therapy were included in the study and received an average of 19.6 months (range: 15.5-33.3 months) of PEG-IFN-2a treatment. At the end of PEG-IFN-2a therapy, 7 (8.6%) of the patients achieved undetectable HBsAg and seroconversion, and 14 (17.3%) showed HBsAg less than 10IU/mL. In addition, 40.7% achieved undetectable HBeAg and seroconversion, a rate that was slightly higher than that (38.3%) seen in treatment-naive patients who received PEG-IFN-2a. Statistical analyses suggest that baseline level of HBsAg at less than 1500 IU/mL may predict end of PEG-IFN-2a treatment response for HBsAg less than 10 IU/mL, as evidenced by the area under the curve measure of 0.747, sensitivity measure of 87.3%, specificity measure of 33.3%, positive predictive value of 82.1% and negative predictive value of 42.8%.</p><p><b>CONCLUSION</b>Patients with HBeAg-positive CHB and partial viral response to NA therapy can achieve undetectable HBsAg and HBeAg seroconversion after switching to PEG-IFN-2a treatment. Baseline HBsAg level may be predictive of response to this therapeutic strategy.</p>

Study on the etiology of acute hepatitis hospitalized patients in Beijing Ditan Hospital from 2002 to 2011 / 中华实验和临床病毒学杂志

<p><b>OBJECTIVE</b>To explore the etiology of acute hepatitis hospitalized patients in Beijing Ditan Hospital from 2002 to 2011.</p><p><b>METHODS</b>We summed up the changes in the characteristics of the etiology of acute hepatitis of patients mentioned above, and preliminarily analyze the causes.</p><p><b>RESULTS</b>From 2002 to 2011, 6235 patients with acute hepatitis were admitted to Ditan Hospital, aged between 12 and 78 years old, Of which 4309 were male and 1926 female. Acute viral hepatitis accounted for 70.44%-85.07%, while CMV, EBV, drug-induced liver injury accounted less than 5%, and acute hepatitis D and acute hepatitis C less than 1.10%. From year to year, the incidence and constitution of acute hepatitis changed significantly. The proportion of patients with acute hepatitis in total hospitalized patients was from 20. 38% to 2.05%. In 10 years, the percentage of acute hepatitis A decreased most obviously, about 99.11%, while 45.07% decline in incidence of acute hepatitis B and 62. 28% of acute hepatitis E. The constituent ratio of acute hepatitis also changed significantly. The proportion of acute hepatitis A declined from 31.31% in 2002, to less than 1% in 2011. The proportion of acute hepatitis B increased from 26.47% in 2002 to 45.88% in 2011, an increase of about 2 folds in 10 years. The proportion of acute hepatitis E increased from 26.73% in 2002 to 32.05% in 2010, a rise of 1.20 times in 10 years.</p><p><b>CONCLUSIONS</b>The proportion of patients with acute hepatitis in total hospitalized patients decreased from 20. 38% in 2002 to 2. 05% in 2011 in Beijing Ditan Hospital. The constituent ratio of acute hepatitis changed, too.</p>

Analyse related factors of impact and prognosis of 73 cases of severe hepatitis / 中华实验和临床病毒学杂志

<p><b>OBJECTIVE</b>A retrospective study was conducted to investigate the clinical features and prognostic factors of 73 cases of severe hepatitis.</p><p><b>METHODS</b>To summarize clinical features of 73 cases of severe hepatitis, grouping by etiology and pathogenesis. A retrospective analysis was performed to evaluate the relationship between biochemical characteristics (liver function, renal function, electrolytes, PTA, etc) and complications (hepatic encephalopathy, upper gastrointestinal bleeding, hepatorenal syndrome, ascites, abdominal infections, etc) and prognosis.</p><p><b>RESULTS</b>(1) HBV infection alone accounted for 65.75%. Alcoholic liver disease, drug-induced liver injury, hepatitis E, autoimmune hepatitis, overlapping causes and other factors were five cases (6.85%), six cases (8.22%), two cases (2.74%), two cases (2.74%), seven cases (9.59%) and three cases (4.11%) respectively. According to the incidence rate, severity and underlying liver condition, subacute hepatitis, cases based on chronic hepatitis and on cirrhosis were 12 cases (16.43%), 11 cases (15.07%), 50 cases (68.49%) respectively. Clinical manifestations with or without hepatic encephalopathy accounted for 58.90% or 41.10%. (2) The highest mortality of severe hepatitis was alcoholic liver disease and patients on the basis of overlapping factors (66.67%), followed by autoimmune liver disease (50%). The mortality of HBV-related hepatitis was 18.75%. Overall mortality of 73 cases of severe hepatitis was 28.77%, of which cirrhosis group was higher than non-cirrhotic group (40% vs 4.3%, P = 0.002). The difference was statistically significant. Patients without hepatic encephalopathy had lower mortality than with hepatic encephalopathy (3.33% vs 46.51%). The mortality of patients with hepatic encephalopathy Stage III and IV was 72.73%. (3) Independent samples t test filtered nine factors associated with death, namely cirrhosis, upper gastrointestinal bleeding, hepatic encephalopathy, hepatorenal syndrome, serum creatinine, total bilirubin (TBIL), direct bilirubin (DBIL), albumin (ALB) and serum sodium. The results of multivariate conditional logistic regression analysis indicated that hepatic encephalopathy, serum creatinine levels were risk factors for death, whereas ALB as a protective factor.</p><p><b>CONCLUSION</b>Hepatic encephalopathy, serum creatinine levels were risk factors for severe hepatitis death, But ALB was protective factor. Nucleotide analogs using was the main reason why the mortality of hepatitis B was as low as 18.75%.</p>

Assemble of magnetic nanoparticles into the structure of cisplatin liposome / 药学学报

Effects of different procedures of magnetic nanoparticles into the liposome structure on the distribution of magnetic particles in the liposome were investigated. Magnetic liposomes with high-encapsulating rate of cisplatin (CDDP) were obtained. Fe3O4 magnetic nanoparticles which was modified by organic functional group on surface was synthesized by an one-step modified hydrothermal method. The CDDP magnetic liposomes were prepared by a film scattering-ultrasonic technique and the concentrations of CDDP in the liposomes were measured by graphite furnace atomic absorbance spectroscopy. Magnetic liposomes with different microstructure were prepared by the two different procedures, where the magnetic particles were combined with phospholipid before the film preparation to form liposome in procedure I, and drug solution and the magnetic particles were mixed before hydrating the lipids film to form liposome in procedure II. The liposome structure was observed by transmission electron microscope (TEM). The CDDP magnetic liposomes were prepared by the optimized method which was selected by orthogonal test. Encapsulation rate of the magnetic particles distributed in the phospholipid bilayer through the procedure I was 34.90%. While liposome, produced by the procedure II technique, contained magnetic particles in the interior aqueous compartment, which encapsulation rate was 28.34%. Encapsulation rates of both I and II were higher than that of conventional liposome. The release profile of all the three different liposomes in vitro fitted with a first-order equation. Because of distribution of magnetic particles in the phospholipid bilayer, the skeleton of phospholipid bilayer was changed. The releasing tl/2 of magnetic liposomes produced by the procedure I technique is 9 h, which is shorter than that of the other two liposomes. Assemble of magnetic nanoparticles into the structure of liposome was succeeded by the procedure I, which showed superiority than by procedure II whatever in CDDP liposome encapsulation efficiency and content of the magnetic particles and would ensure sustained-release character.

Vasorelaxational effects of procyanidins on rabbit aorta in vitro and decreasing arterial blood pressure in vivo / 中国中药杂志

<p><b>OBJECTIVE</b>To study the vasodilation effect of the procyanidin (PC) extracted from grape seeds on rabbit thoracic aortic rings in vitro, decreasing blood pressure in vivo and the possible mechanism.</p><p><b>METHOD</b>Rabbits aortic rings were isolated and were divided into six groups including removal of endothelium, integrity of endothelium, 1 x 10(-5) mol X L(-1) indomethacin (Indo), 1 x 10(-5) mol x L(-1) propranolol (Prop), 1 x 10(-4) mol x L(-1) N(omega)-nitro-L-arginine (L-NNA) and 1 x 10(-5) mol x L(-1) methylene blue (MB). Then the thoracic aortic rings were treated with PC with cumulative concentrations of 1.25, 2.5, 5.0 mg x L(-1) respectively and the changes of tension were recorded, and investigate the effect of 40 mg x L(-1) PC on the contraction of aortic smooth muscles, thoracic aortic rings were pre-treated with NA (1 x 10(-8) to approximately 1 x 10(-5) mol x L(-1)), KCl (6.3 to approximately 100 mmol x L(-1)) and CaCl2 (1 x 10(-5) to approximately 1 x 10(-5) mol x L(-1)) followed by treatment with PC. Then, rabbits common carotid artery was intubated and arterial blood pressure in vivo was recorded. PC with cumulative concentrations of 4.0, 8.0, 16, 32, 64, 84 mg x kg(-1) was injected into vein and the changes of arterial blood pressure were observed.</p><p><b>RESULT</b>PC could relax isolated rabbit aorta and showedan obvious concentration-dependent relaxation (r = 0. 63, P < 0.001). The relaxant effect of PC was significantly reduced by removal of endothelium and by treatment with nitric oxide synthase inhibitor L-NNA, or guanylyl cyclase inhibitor MB. In addition PC could decrease the dose response curves of aortic rings to NA, KCl and CaCl2. PC has a significant concentration-dependent negative effect on arterial blood pressure in vivo (r = 0.92, P < 0.001).</p><p><b>CONCLUSION</b>PC has a vasodilation effect not only in an endothelium-dependent, nitric oxide involved manner, but in inhibition of calcium release and blockage of potential-dependent calcium channels. PC could decrease the rabbit's arterial blood pressure significantly in vivo.</p>

Effect of cultivating conditions on alpha-galactosidase production by a novel Aspergillus foetidus ZU-G1 strain in solid-state fermentation / 浙江大学学报（英文版）（B辑：生物医学和生物技术）

The work is intended to achieve optimum culture conditions of alpha-galactosidase production by a mutant strain Aspergillus foetidus ZU-G1 in solid-state fermentation (SSF). Certain fermentation parameters involving moisture content, incubation temperature, cultivation period of seed, inoculum volume, initial pH value, layers of pledget, load size of medium and period of cultivation were investigated separately. The optimal cultivating conditions of alpha-galactosidase production in SSF were 60% initial moisture of medium, 28 degrees C incubation temperature, 18 h cultivation period of seed, 10% inoculum volume, 5.0 approximately 6.0 initial pH of medium, 6 layers of pledget and 10 g dry matter loadage. Under the optimized cultivation conditions, the maximum alpha-galactosidase production was 2 037.51 U/g dry matter near the 144th hour of fermentation.