1.Preparation and immunogenicity evaluation of recombinant poliomyelitis type 2 virus-like particles
Yang XU ; Zhifang YING ; Lin XU ; Jianfeng WANG ; Yueyue LIU ; Hongyan LI ; Qiqi HAN ; Qiaoling YAN ; Tao ZHU
Chinese Journal of Microbiology and Immunology 2021;41(10):784-790
Objective:To express virus-like particles of poliovirus type 2 (PV2-VLP) in insect cells using a recombinant baculovirus expressing P1 and 3CD and to preliminarily evaluate its immunogenicity.Methods:Based on the codon preference of High 5 cells, the sequences of P1 gene and 3CD gene of PV2 were optimized and inserted into pUC57-Amp to construct pUC57-PV2-P1 and pUC57-PV2-3CD. UC57-PV2-P1s mutant that carried P1 gene mutation affecting thermostability was then constructed. Recombinant baculovirus strains of rBac-PV2-P1s-3CD and rBac-PV2-P1-3CD (wild type) were constructed using homologous recombination. The expression of target proteins was detected by Western blot. PV2-VLP was purified by ion exchange chromatography. The structure of VLP was observed under transmission electron microscopy to evaluate the assembly efficiency. The immunogenicity of PV2-VLP was assessed in a rat model.Results:The recombinant baculovirus with stable expression of P1s and 3CD proteins was successfully constructed. Western blot results showed that the yield of VLP was higher after thermostability mutation than that of the wild type. A three-dimensional structure with a diameter of about 30 nm was observed under electron microscopy, indicating that the VLP was successfully assembled. Animal experiment showed that the recombinant PV2-VLP had immunogenicity and could effectively induce the production of neutralizing antibodies.Conclusions:Effective VLP vaccines could be successfully prepared using the insect cell-baculovirus expression system, which provided reference for the development of polio VLP vaccine.
2.A multicenter survey of short-term respiratory morbidity in late-preterm infants in Beijing
Tongyan HAN ; Xiaomei TONG ; Xin ZHANG ; Jie LIU ; Li YANG ; Hui LIU ; Ju YAN ; Zhifang SONG ; Yabo MEI ; Xiaojing XU ; Rong MI ; Xuanguang QIN ; Yuhuan LIU ; Yujie QI ; Wei ZHANG ; Huihui ZENG ; Hong CUI ; Hui LONG ; Guo GUO ; Xulin CHEN ; Zhaoyi YANG ; Fang SUN ; Changyan WANG ; Zhenghong LI
Chinese Journal of Applied Clinical Pediatrics 2020;35(16):1230-1234
Objective:To study the respiratory morbidity and the risk factors of respiratory complications in late-preterm infants.Methods:The data of 959 late-preterm infants in 21 hospitals in Beijing from October 2015 to April 2016 were collected.These infants were divided into the respiratory morbidity group (237 cases) and the control group (722 cases) according to whether they had short-term respiratory morbidity after birth.Clinical data of the two groups were compared.Results:Among the 959 late-preterm babies, 530 were male and 429 were female.Two hundred and thirty-seven cases (24.7%) developed short-term respiratory morbidity after birth.Infectious pneumonia developed in the most cases (81 cases, 8.4%), followed by transient tachypnea (65 cases, 6.8%), amniotic fluid aspiration (51 cases, 5.3%), and respiratory distress syndrome (24 cases, 2.5%) successively.All the infants recovered and discharged.There were no differences between gender and maternal age between 2 groups (all P>0.05). Compared with the control group, more late-preterm infants were delivered by cesarean section (73.4% vs.59.7%, χ2=14.43, P<0.001) and the 1-minute Apgar score was lower [(9.41±1.66) scores vs.(9.83±0.53) scores, t=5.40, P<0.001] in the respiratory morbidity group.The differences were statistically significant.There were more cases with maternal complications in the respiratory morbidity group that in the control group (66.7% vs.58.6%, χ2=4.877, P=0.027), but no difference in various complications between 2 groups was observed ( P>0.05). In the respiratory morbidity group, the most frequent complications were maternal hypertension and preeclampsia (27.8% vs.22.6%, χ2=2.728, P=0.099). There were no differences between 2 groups in gestational age, birth weight and birth length (all P>0.05). There were more infants small for gestational age and large for gestational age in the respiratory morbidity group than in the control group (18.8% vs.14.1%, 6.3% vs.2.4%, χ2=8.960, P=0.011). The duration of hospitalization of the respiratory morbidity group was significantly longer than that of the control group [(9.00±4.42) d vs.(6.82±4.19) d, t=6.676, P<0.001] since the infants with respiratory morbidity needed to be hospita-lized. Conclusions:Respiratory diseases occur in about 1/4 of late-preterm infants.Infants who are delivered by cesarean section and whose mothers are complicated with the maternal hypertension and preeclampsia should be monitored closely.Respiratory support should be provided for infants not appropriate for gestational age who are more likely to suffer from respiratory diseases, so that they can successfully pass through the transition period.
3.A multi-center research on risk factors of hyperbilirubinemia in late preterm infants
Xiaochun CHEN ; Li YANG ; Huihong ZHU ; Xin ZHANG ; Jie LIU ; Tongyan HAN ; Hui LIU ; Jü YAN ; Zhifang SONG ; Yabo MEI ; Xiaojing XU ; Rong MI ; Xuanguang QIN ; Yuhuan LIU ; Yujie QI ; Wei ZHANG ; Huihui ZENG ; Hong CUI ; Changyan WANG ; Zhenghong LI ; Hui LONG ; Guo GUO ; Xulin CHEN ; Zhaoyi YANG ; Fang SUN
Chinese Journal of General Practitioners 2018;17(12):992-996
Objective To investigate the risk factors of hyperbilirubinemia in late preterm infants. Methods The clinical data of 815 late preterm infants (449 males and 366 females) from 25 hospitals in Beijing were collected from October 2015 to April 2016, including 340 cases(41.7%) with hyperbilirubinemia (hyperbilirubinemia group), and 475 cases without hyperbilirubinemia (control group). The clinical data of two groups were compared, and the maternal factors influencing hyperbilirubinemia in late preterm infants were analyzed with logistic regression. Results There were no significant differences in gender ratio (M:F 1.39 vs. 1.12, t=1.811,P=0.172)and birth weight[(2502.6±439.6)g vs. (2470.2±402.9)g,χ2=2.330,P=0.127)]between two groups. The incidence rates of hyperbilirubinemia in infants of 34 wks, 35 wks and 36 wks of gestational age were 22.9%(87/174), 35%(119/300) and 42.1%(143/341) respectively (χ2=1.218,P=0.544). The multivariate logistic regression analysis indicated that the maternal age(OR=1.044,95% CI:1.010-1.080,P=0.011)was independent risk factor and multiple births(OR=1.365,95%CI:0.989-1.883,P=0.048), premature rupture of membranes(OR=2.350,95% CI:1.440-3.833,P=0.001), cesarean section(OR=1.540,95%CI:0.588-4.031,P=0.014)were risk factors for hyperbilirubinemia in late preterm infants. Conclusions The incidence of hyperbilirubinemia in late preterm infants is relatively high. Maternal age, multiple births, premature rupture of membranes and cesarean section are risk maternal factors related to hyperbilirubinemia in late preterm infants.
4.Release Mechanism and Preparation of Liposome Gel of Total Alkaloids of Sophoras Flavescentis Radix
Lingyin ZHU ; Guantao ZHENG ; Changni ZHOU ; Yanli SHEN ; Zhifang HAN ; Yongyan JIA
Chinese Journal of Information on Traditional Chinese Medicine 2017;24(1):77-81
Objective To optimize the formulation and process of liposome gel of total alkaloids of Sophoras Flavescentis Radix; To prepare the liposome gel of total alkaloids of Sophoras Flavescentis Radix and study its release mechanism.Methods Matrine liposomes was prepared by using film dispersion method; With entrapment efficiency and medicine loading as indexes, acid dye colorimetric method was used for the determination of matrine content in liposomes. Orthogonal design was used to optimize the formulation and the optimal formulation of liposomes was selected. Poloxamer-407 was set as the substrate preparation of matrine liposome gel. The transdermal rate of medicine gel and medicine liposome gel was investigated.Results Obtained through formulation and technology optimization of liposomes formation uniform, particle size was in the range of 100 nm to 400 nm, entrapment 74%, loading 26%. Preparation of liposome gel was transparent semisolid. In vitro results showed, cumulative release dose of matrine hydrogel was 6.34 mg/cm2 within 48 h; cumulative release doses of liposome gel of total alkaloids of Sophoras Flavescentis Radix was 6.97 mg/cm2 within 48 h; cumulative volume, steady-state penetration rate through skin and 48 h volume in the skin of the latter were significantly improved compared with that of the former. Conclusion Optimum preparation is reliable and practical. Liposome gel of total alkaloids of Sophoras Flavescentis Radix made by the preparation is with high quality, which can effectively delay the medicine release rate, increase the volume of medicine in human body.
5.Investigation and Analysis of Internships and Employment Intention of Pharmacy Junior Students in Our University
Zhuo DU ; Feidie LIU ; Jianfeng WU ; Haiyan LI ; Xin NIE ; Zhifang CEN ; Weiguang LIANG ; Aifen YAN ; Bin HAN ; Zhong LI
China Pharmacy 2017;28(12):1588-1591
OBJECTIVE:To provide evidence for strengthening education and training before clinical practice and employment guidance among pharmacy junior students of our university. METHODS:Anonymous questionnaire survey was conducted for our university. Questionnaire survey included student employment intention and internship units,relationship of internships with employ-ment,the tendency of employment after graduation,personal attitude about pharmaceutical professional development prospect,ex-pected monthly income in the first year of their career,etc. RESULTS:Totally 71 questionnaires were sent out,71 were effectively received. Among all respondents,74.3%(52 students)believed employment intention had great relationship with internship units;24.3%(17 students)thought employment intention had not great relationship with internship units;moreover,one student thought employment intention had no any relationship with internship units. Most students(94.3%,66 students)with good attitude on in-ternships believed it could accumulate work experience;32.9%(23 students) thought they could stay in the internship unit after practice;moreover,17.1%(12 students) believed practice had little effect on employment. Among them,94.0%(63 students) tended to be employed after graduation,and only 6.0% chose a graduate school to continue their studies. Among the students who chose employment after graduation,76.1%(51 students)tended to engage in pharmaceutical related work,while 17.9%(12 stu-dents) tended to be engaged in work nothing to do with pharmacy or self-employed. The majority (69.6%,48 students) believed that pharmaceutical major prospect was general and pharmaceutical major development varied from person to person;17.4%(12 students)thought pharmaceutical major had good prospect and was promising;13.0%(9 students)believed that pharmaceutical ma-jor had no good prospect and didn't know its prospect. Most of the students(78.2%,54 students)expected a monthly income of 2500-5000 yuan in the first year of their career;18.9%expected a monthly income more than 5000 yuan(13 students);the minor-ity expected a monthly income of 1500-2500 yuan or more than 8000 yuan. CONCLUSIONS:The view of students on internship and employment have a certain gap with the social situation. Related departments of colleges and universities need to adjust the thought of students and strengthen guidance.
6.Effect of contracted learning on rehabilitation training to patients with stroke
Binying HUANG ; Zhifang HUANG ; Guang YANG ; Hua LIU ; Xiaojing ZHU ; Tianrong HUANG ; Weili HAN ; Wenzhong JIANG ; Yajie LI
Modern Clinical Nursing 2016;15(5):26-30
Objective To explore the effect of contract learning on rehabilitation training to patients with stroke . Methods According to the random number table, 120 stroke patients were randomly divided in equal number into control group and observation group. The patients in the control group were treated with conventional nursing during hospitalization and health guidance during follow-up. The patients in the observation group were given rehabilitation training according to contracted learning during the period of hospitalization and follow-up. The time was 6 months. The quality of life was compared between the two groups of patients before and after the intervention, 1 months after discharge, 6 months after discharge. Results After six months of intervention, the 7 dimensions of quality of life, including daily life ability, hand function, mobility, strength, emotion, communication and participation in the intervention group were significantly better than those of the control group (P<0.001). The 6 dimensions of quality of life, including daily life ability, hand function, mobility, strength, emotion, and communication of the control group were significantly better than before the intervention (P<0.001). Conclusion For stroke patients, the rehabilitation training under the learning contract can improve the effect of rehabilitation training so as to improve the quality of life and the prognosis.
8.Expression and significance of fat mass and obesity associated gene and forkhead transcription factor O1 in non-alcoholic fatty liver disease.
Jielei ZHANG ; Shan LI ; Jingyi LI ; Chao HAN ; Zhifang WANG ; Chong LI ; Xiaoman WANG ; Zhenzhen LIU ; Jianguo WEN ; Lili ZHENG
Chinese Medical Journal 2014;127(21):3771-3776
BACKGROUNDNon-alcoholic fatty liver disease (NAFLD) is a complex disorder and has been closely linked to obesity. The fat mass and obesity-associated (FTO) gene is a newly discovered gene related to obesity, which enhances oxidative stress and lipogenesis in NAFLD. The forkhead transcription factor O1 (FoxO1) is another important gene involved in NAFLD, which causes lipid disorders when insulin resistance appears in the liver. However, the interactions between FTO and FoxO1 during the pathogenesis of NAFLD have not been fully elucidated. This study was designed to identify the relationship between these two factors that are involved in the development of NAFLD.
METHODSThis study includes two parts referred to as animal and cell experiments. Twelve female SPF C57BL/6 mice were fed a high-fat diet to serve as an NAFLD animal model. Aspartate aminotransferase (AST), alanine aminotransferase (ALT), total triglyceride (TG), total cholesterol (TC), alkaline phosphatase (ALP), high-density lipoprotein (HDL), and low-density lipoprotein (LDL) were measured. Immunohistochemical analysis was used to detect the expression and histological localization of FTO, FoxO1, and adenosine monophosphate (AMP)-activated protein kinase (AMPK). The L02 cells were exposed to high fat for 24, 48, or 72 hours. Oil red O staining was used to detect intracellular lipid droplets. Reverse transcription-polymerase chain reaction was used for analyzing the levels of FTO and FoxO1 mRNA.
RESULTSAt the end of 10 weeks, ALP, ALT, AST, and LDL were significantly increased (P < 0.01), while TC and TG were also significantly higher (P < 0.05). In addition, HDL was significantly decreased (P < 0.05). The FTO and FoxO1 proteins were weakly expressed in the control group, but both FTO and FoxO1 were expressed significantly higher (P < 0.01) in the experimental group, and the expression of the two factors was significantly correlated. AMPK in the high-fat group showed a low level of correlation with FTO, but not with FoxO1. Oil Red O staining results showed that the cells cultured in 50% fetal bovine serum for 24, 48, or 72 hours exhibited steatosis. FTO and FoxO1 mRNA were increased in the high-fat group compared with the normal group (P < 0.01). The expression levels of FTO and FoxO1 mRNA were the highest at 48 hours (P < 0.05).
CONCLUSIONSA high-fat diet leads to higher expression of FTO, phosphorylation of FoxO1, and decreased phosphorylation of AMPK. These results suggest that the interactions between FTO and FoxO1 are closely related to the pathogenesis of NAFLD.
Alanine Transaminase ; genetics ; metabolism ; Animals ; Aspartate Aminotransferases ; genetics ; metabolism ; Cholesterol ; metabolism ; Diet, High-Fat ; adverse effects ; Disease Models, Animal ; Female ; Forkhead Transcription Factors ; genetics ; metabolism ; Lipoproteins, HDL ; metabolism ; Lipoproteins, LDL ; metabolism ; Liver ; metabolism ; Mice ; Mice, Inbred C57BL ; Non-alcoholic Fatty Liver Disease ; metabolism ; pathology ; Obesity ; metabolism ; pathology ; Triglycerides ; metabolism
9.Causes analysis of unexpected extubation of 1 0 patients in hepatobiliary surgery and its preventive nursing
Xianghua HAN ; Ruilan WANG ; Qiuxia HUANG ; Zhifang BAI ; Hongwei TAO
Journal of Clinical Medicine in Practice 2014;(18):43-45,52
Obj ective To explore yhe causes and yhe nursing measures of unexpecyed exyu-bayion of indwelling cayheyer in hepayobiliary surgery.Methods The clinical daye of 10 payienys wiyh unexpecyedly exyubayion among 402 payienys wiyh hepayobiliary surgery and indwelling cayheyers was analyzed.Results In 10 unexpecyedly exyubayion payienys,yhere were 3 payienys pulling ouy yhe syomach yube by yhemselves,2 payienys pulled ouy abdominal drainage yube in yhe nighy when yurning over.2 payienys′abdominal drainage were auyomayically pulled ouy.1 payieny′s cayheyer and 1 payieny’s deep venous inyubayion were pulled ouy by overly power in yheir yranspory. Jejunum nuyriyion yube wiyhouy suyure fixayion of 1 payieny was pulled ouy.Conclusion According yo yhe causes of unexpecyed exyubayion,improvemeny of nursing measures,syandardized and insyiyu-yionalized nursing can reduce yhe incidence raye of unexpecyed exyubayion.
10.Expression and significance of fat mass and obesity associated gene and forkhead transcription factor O1 in non-alcoholic fatty liver disease
Jielei ZHANG ; Shan LI ; Jingyi LI ; Chao HAN ; Zhifang WANG ; Chong LI ; Xiaoman WANG
Chinese Medical Journal 2014;(21):3771-3776
Background Non-alcoholic fatty liver disease (NAFLD) is a complex disorder and has been closely linked to obesity.The fat mass and obesity-associated (FTO) gene is a newly discovered gene related to obesity,which enhances oxidative stress and tipogenesis in NAFLD.The forkhead transcription factor O1 (FoxO1) is another important gene involved in NAFLD,which causes lipid disorders when insulin resistance appears in the liver.However,the interactions between FTO and FoxO1 during the pathogenesis of NAFLD have not been fully elucidated.This study was designed to identify the relationship between these two factors that are involved in the development of NAFLD.Methods This study includes two parts referred to as animal and cell experiments.Twelve female SPF C57BL/6 mice were fed a high-fat diet to serve as an NAFLD animal model.Aspartate aminotransferase (AST),alanine aminotransferase (ALT),total triglyceride (TG),total cholesterol (TC),alkaline phosphatase (ALP),high-density lipoprotein (HDL),and low-density lipoprotein (LDL) were measured.Immunohistochemical analysis was used to detect the expression and histological localization of FTO,FoxO1,and adenosine monophosphate (AMP)-activated protein kinase (AMPK).The L02 cells were exposed to high fat for 24,48,or 72 hours.Oil red O staining was used to detect intracellular lipid droplets.Reverse transcription-polymerase chain reaction was used for analyzing the levels of FTO and FoxO1 mRNA.Results At the end of 10 weeks,ALP,ALT,AST,and LDL were significantly increased (P <0.01),while TC and TG were also significantly higher (P <0.05).In addition,HDL was significantly decreased (P <0.05).The FTO and FoxO1 proteins were weakly expressed in the control group,but both FTO and FoxO1 were expressed significantly higher (P <0.01) in the experimental group,and the expression of the two factors was significantly correlated.AMPK in the high-fat group showed a low level of correlation with FTO,but not with FoxO1.Oil Red O staining results showed that the cells cultured in 50% fetal bovine serum for 24,48,or 72 hours exhibited steatosis.FTO and FoxO1 mRNA were increased in the high-fat group compared with the normal group (P <0.01).The expression levels of FTO and FoxO1 mRNA were the highest at 48 hours (P <0.05).Conclusions A high-fat diet leads to higher expression of FTO,phosphorylation of FoxO1,and decreased phosphorylation of AMPK.These results suggest that the interactions between FTO and FoxO1 are closely related to the pathogenesis of NAFLD.

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