1.Isolation of the variable fragments of heavy chain against the terminal protein region of hepatitis B virus polymerase with protein fragment complementation assay
Junyan YU ; Lin LAN ; Yuming WANG ; Shitao DING
Chinese Journal of Immunology 1985;0(06):-
Objective:To isolate the variable fragments of heavy chain(VH) against the terminal protein(TP) region of hepatitis B virus(HBV) DNA polymerase(Pol) with protein fragment complementation assay(PCA).Methods:The TP region of HBV secreted by the HepG2.2.15 cells was used as an antigen,and the antibodies were selected with PCA.In this assay,two interacting proteins(target and antibody) are genetically fused to the two halves of the dissected enzyme dihydrofolate reductase.Binding of the two partners reassembles this enzyme and reconstitutes its activity,thus allowing growth on minimal mediem.Results:There were three TP region antigen-specific VHs could be directly in vivo selected with PCA.Sequence analysis showed that each TP-specific VH had a different sequence.Conclusion:Our results show that TP region antigen-specific VH could be directly in vivo selected with PCA.This system were powerful as a routine system for generating antibodies,especially in functional genomics.
2.Development of Simple Photographic Device for Field Battle
Fanju PAN ; Chuangao YANG ; Shitao QI ; Baojun WANG ; Fengqin XIN
Chinese Medical Equipment Journal 1993;0(05):-
Objective To develop a simple new-type photographic device for the field battle and peace time,because of the function of the trivet of X-Ray machine is limited.Methods Circular chrome-plating seamless steel tubes,whose diameter is 4 cm and 5 cm respectively,were connected so as to constitute the arm structure in C shape and foundation in T shape.Results The simple new-type photographic device could project for patients in each kind of body posture and the range of focus adjustment could meet normal requirements.It was operated nimbly and freely.The material was durable and the structure was reasonable and stable.Conclusion The simple new-type photographic device meets the practical requirements in field battle and peace time.
3.Clinical observation after Roux-en-Y gastric bypass in type 2 diabetes mellitus
Shitao HU ; Ruixiang DUAN ; Yong ZHANG ; Dong WANG ; Linlin XU ; Xiaoli WANG ; Hong JI ; Haiqin RONG
The Journal of Practical Medicine 2014;(5):792-795
Objective To observe multiple metabolic changes within one year after Roux-en-Y gastric bypass (RYGB) surgery in obese patients with type 2 diabetes mellitus, and to evaluate the index of the gastric bypass surgery and the determination of applicable population. Methods The clinical and laboratory data before and after surgery in 112 patients diagnosed as type 2 diabetes mellitus by RYGB were included in this study. According to BMI, these patients were divided into two groups: normal group (≤ 25kg/m2), and the overweight group (> 25 kg/m2). The physiologic and biochemical indexes of the patients were followed up at 1, 3, 6, 12 months, respectively. The statistical analysis was performed by SPSS17.0 software. Results Compared with the preoperative period, the levels of HbA1c and HOMA-IR in the postoperative period were significantly increased in the two groups. Principal component analysis showed that the postoperatively contributions of BMI and HbA1c in 6 months was bigger than that of the other indexes, while HOMA-β, HOMA-IR had larger contributions after 6-month postoperative period. Conclusion Various metabolic indexes in patients with type 2 diabetes improve significantly within one year after surgery, and the complete remission rate is gradually increased. The indexes including weight, blood glucose, serum lipids, HOMA-βand HOMA-IR in overweight and obese group have more significant improvements than those in normal group. Obese patients are more appropriate for the surgical treatment. The indexes, such as BMI, HbA1c,HOMA-βand HOMA-IR, should help to evaluate the operating effectiveness and preoperative indications.
4.Study on Curing Time Controlling of Materials Based on Isocyanate and Its Application in Transmitting Apparatus for the Wounded
Tiebing LIU ; Liming TANG ; Zheng WANG ; Shengjun LIU ; Min WU ; Shitao QI ; Xingxi ZHU ; Chao ZHANG
Chinese Medical Equipment Journal 2009;30(7):21-24
Obiective To study controUing method for curing time of the materials based on isocyanate, and perfect making means and operating rules of transmitting apparatus for the wounded, Methods According to the controlling principles of the rate of chemical reaction, the influencing factors' were studied such as catalyzer, ambient temperature, mixture level of materials and materials volume by experiment. Results By the results of experiments, ambient temperature hasn't an effector impact on eufing time of isocyanate, catalyzer and mixture level of materials only have a little effect or impact on initial reaction time, but materials volume is an important factor of all. Conclusion By the means of experiments, the curing time of isocyanate is only influenced by the volume of materials, and the character is very useful to transmitting apparatus for the wounded in field battle or on-site emergency rescue.
5.Clinical analysis of mitochondrial encephalomyopathy with lactic acidosis and stroke -like episodes in children
Shitao WU ; Hengfang LIU ; Fang LIU ; Min ZHANG ; Yuanzheng ZHAO ; Yapei GUO ; Ningning CHEN ; Jianping WANG
Chinese Journal of Applied Clinical Pediatrics 2016;31(24):1884-1888
Objective To investigate the clinical,imaging,pathological and molecular biological features of mitochondrial encephalomyopathy with lactic acidosis and stroke -like episodes(MELAS)in children.Methods The clinical,imaging,pathological and molecular biological features of 1 2 children with MELAS diagnosed through muscle biopsy or gene sequencing in the Fifth Affiliated Hospital of Zhengzhou University from January 201 1 to December 201 5 were retrospectively analyzed.Results (1 )Clinical features:the main manifestations included headache and vomiting in 1 1 cases,epileptic seizures in 9 cases,short stature in 8 cases,hairy in 7 cases,intolerance fatigue in 7 cases,cogni-tive decline in 7 cases,visual disturbance in 6 cases,hearing disturbance in 6 cases,and 5 cases had positive family history.In addition,7 cases had the serum lactic acid level increase in a rest for 1 0 min after exercise.(2)Imaging fea-tures:4 cases showed bilateral basal ganglia calcification symmetry in 8 patients who underwent head CT scan.The most frequently involved parts of the lesion were occipital in 1 0 cases,temporal in 9 cases and parietal lobe in 7 cases in stroke -like episodes.The lesions were lamellar necrosis.The abnormal areas by MRI showed low signal intensity on T1 weighted imaging,high signal intensity on T2 weighted imaging and fluid attenuated inversion recovery,high or equal signal intensity on diffusion weighted imaging,high or low signal intensity on apparent diffusion coefficient;the lactate peak significantly increased on magnetic resonance spectroscopy.The distribution was not in accordance with the control region of the cerebral vessels.Dynamic observation revealed that the lesions were reversible and migratory.(3)Myo-pathological features:muscle biopsy was performed in all children,and ragged -red fibers were found in 1 0 cases by im-proved Gomori staining,strongly succinate dehydrogenase -reactive were found in 9 cases,and the lipid droplets slight-ly increased in 8 cases by oil red O staining.Besides,the crystalline inclusion bodies in mitochondria were arranged in a parking lotpattern in 9 cases by electromicroscope.(4)Molecular biological characteristics:the mitochondrial gene mutations were analyzed in peripheral blood of 9 children and their parents,including 8 cases with A3243G muta-tion and 1 case with G13513A mutation.Five mothers had the same A3243G mutation site in 8 cases.Conclusions Children with MELAS have complex and varied clinical manifestations and certain characteristic of neuroimaging.More-over,muscle pathology and gene sequencing have important diagnostic value.Fully understanding the clinical,muscle pathology,imaging and molecular biological characteristics of children with MELAS can be helpful to the early diagnosis and treatment,also reduce misdiagnosis.
6.Recent advance in roles of brain-derived neurotrophic factor-tyrosine receptor kinase B signaling pathway genes and their encoded proteins in epilepsy
Shitao WANG ; Jin LONG ; Liang ZHOU ; Yanbing HAN
Chinese Journal of Neuromedicine 2020;19(12):1269-1272
Brain-derived neurotrophic factor (BDNF) and tyrosine receptor kinase B (TrkB) signaling pathway may play important roles in a variety of neurological diseases, particularly in the development and treatment of epilepsy. The BDNF gene and NTRK2 gene are genes encoding BDNF and TrkB proteins in this pathway. The variations in these two genes may affect the occurrence and treatment of epilepsy through leading to abnormal functions and expressions of the encoded proteins, which may affect the degrees of activation of BDNF-TrkB signaling pathway. This article summarizes the pathophysiological processes of epilepsy in which BDNF-TrkB signaling pathway genes and their encoded proteins may be involved.
7.Mises stress analysis of 42A2 type tibial mid segment oblique fracture fixed with external locking plate at different angles
Yunqi YANG ; Xiaoye WANG ; Lei MI ; Guanjun ZHANG ; Ling ZHANG ; Shitao FANG ; Yun WANG
Journal of Chinese Physician 2023;25(5):700-703,708
Objective:To establish a three-dimensional model of locking plate fixation for 42A2 type oblique tibial fractures with different fracture line directions and different angles between the fracture line and the long axis of the tibia. Finite element analysis was used to calculate and analyze the biomechanics of locking plate, screw, and tibia, providing theoretical basis for clinical application.Methods:A healthy adult volunteer, 25 years old, male, with a height of 173 cm and a weight of 69.5 kg, was selected to perform computed tomography (CT) scans on the left tibia. Relevant data were obtained to establish a locking steel plate fixation model for 42A2 type tibia with different oblique fracture line directions and different angles between the fracture line and the long axis of the tibia. Eight hole pure titanium plates were used for fixation, respectively. We compared the Mises stress changes of locking plates, screws, and tibia in different angle fracture models.Results:In the case of a 42A2 type fracture in the left oblique direction with a fracture line from outside to inside, the maximum Mises stress in the tibia was 114 MPa, the maximum Mises stress in the screw was 279.8 MPa, and the maximum Mises stress in the locking steel plate was 302.4 MPa; In the case of a 42A2 type fracture in the right oblique fracture with a fracture line from the bottom to the top, the maximum Mises stress of the tibia was 93.41MPa, the maximum Mises stress of the screw was 353.4 MPa, and the maximum Mises stress of the locking steel plate was 411.8 MPa.Conclusions:Regardless of the oblique fractures in both left and right directions, the maximum stress values are: locking plate>screw>tibia; When the position of the locking steel plate is fixed, the maximum stress values of the locking steel plate and screw are both right oblique fracture>left oblique fracture; And the maximum stress values all increase with the increase of angle.
8.Study on Compound Musk in the treatment of hypertensive cerebral hemorrhage after operation
Baoyi YANG ; Xiangmei YE ; Jianwei WANG ; Shitao XIA
Chinese Journal of Postgraduates of Medicine 2023;46(3):199-204
Objective:To investigate the efficacy of Compound Musk combined with nimodipine combined with minimally invasive surgery in the treatment of hypertensive cerebral hemorrhage and the effects on serum inflammation, stress and apoptosis.Methods:Prospective research methods was used. A total of 118 patients with hypertensive intracerebral hemorrhage who received treatment in the First Affiliated Hospital of Heilongjiang University of Traditional Chinese Medicine from March 2017 to January 2021 were randomly divided into control group and Compound Musk group (59 cases in each group). After minimally invasive surgery, patients in the control group were treated with nimodipine on the basis of conventional treatment, while patients in the Compound Musk group were treated with compound musk on the basis of the control group. After 2 weeks, the efficacy was evaluated and the levels of serum inflammatory indexes, oxidative stress indexes and apoptosis indexes were measured.Results:The total effective rate in Compound Musk group was higher than that in control group: 98.3% (58/59) vs. 88.1% (52/59), and the difference was statistically significant ( P<0.05). After 2 weeks of treatment, serum inflammatory indexes including nuclear factor-κB (NF-κB), interleukin-1β (IL-1β), matrix metalloproteinase-3 (MMP-3), matrix metalloproteinase-9 (MMP-9); apoptosis indexes including soluble Fas receptor (sFas), soluble Fas ligand (sFAS-L); oxidative stress indexes including advanced oxidation protein products (AOPP), malondialdehyde (MDA) decreased, and some oxidative stress indexes including glutathione peroxidase (GSH-Px), catalase (CAT) increased. The levels of the above inflammatory indexes, apoptosis indexes and oxidative stress indexes in Compound Musk group were lower than those in control group, NF-κB: (18.96 ± 2.17) ng/L vs. (24.10 ± 3.23) ng/L, IL-1β: (12.88 ± 1.74) ng/L vs. (15.19 ± 1.63) ng/L, MMP-3: (5.62 ± 0.95) ng/L vs. (7.97 ± 0.86) ng/L, MMP-9: (7.07 ± 0.86) ng/L vs. (9.26 ± 1.13) ng/L, sFas: (3.24 ± 0.38) μg/L vs. (4.19 ± 0.53) μg/L, sFas-L: (209.17 ± 24.39) ng/L vs. (288.54 ± 37.61) ng/L, AOPP: (10.76 ± 1.84) μg/L vs. (13.51 ± 2.09) μg/L, MDA: (2.87 ± 0.32) μmol/L vs. (3.45 ± 0.34) μmol/L, and the differences were statistically significant ( P<0.05). Some of the above oxidative stress indexes were higher than those in control group, GSH-Px: (3 274.91 ± 376.09) U/L vs. (2 854.19 ± 325.22) U/L, CAT: (60.82 ± 7.43) U/L vs. (52.17 ± 6.48) U/L, the differences were statistically significant ( P<0.05). During treatment, there was no significant difference in the incidence of rash, diarrhea, drug-induced liver and myocardial injury between two groups ( P>0.05). Conclusions:Compound Musk has a positive effect on improving the curative effect and internal environment of patients with hypertensive intracerebral hemorrhage after minimally invasive surgery, and will not increase the occurrence of serious adverse reactions.
9.Exploring Synergistic Mechanism of Vinegar-processed Olibanum in Treatment of Ulcerative Colitis Based on Bile Acid-regulated "Gut-liver" Crosstalk
Shitao PENG ; Zhenli LIU ; Zhiqian SONG ; Chun WANG ; Zhangchi NING
Chinese Journal of Experimental Traditional Medical Formulae 2023;29(21):59-68
ObjectiveTo investigate the synergistic mechanism of vinegar-processed Olibanum on ulcerative colitis(UC) via the bile acids regulating "gut-liver" crosstalk. MethodRats were randomly divided into normal group, model group, Olibanum group and vinegar-processed Olibanum group. UC model of rats was induced by intracolonic instillation of 2,4,6-trinitrobenzenesulfonic acid(TNBS). Ultra high performance liquid chromatography-triple quadrupole-mass spectrometry(UPLC-QQQ-MS) was used to perform the qualitative analysis of 30 bile acids in the colon of rats. Real-time fluorescence quantitative polymerase chain reaction(Real-time PCR) and Western blot were used to detect changes in the expression of farnesoid X receptor(FXR), fibroblast growth factor 15(FGF15) and FGF receptor 4(FGFR4) in "gut-liver" crosstalk at mRNA and protein levels. And with the help of HcoEpiC cell model intervened by conjugated bile acids, simulating the UC state, and according to the different modes of intervention, they were divided into the blank group, conjugated bile acid group, Olibanum group, vinegar-processed Olibanum group and 3-O-acetyl-9,11-dehydro-β-boswellic acid(ADHBA) group. The effect of Olibanum before and after processing with vinegar and the main differential component ADHBA on the mRNA expression of FXR and FGF19 were explored by Real-time PCR. ResultCompared with the normal group, the levels of conjugated bile acids in the model group increased significantly(P<0.01), and the mRNA and protein expressions of "gut-liver" crosstalk factors FXR, FGF15 and FGFR4 decreased significantly(P<0.05, P<0.01). Compared with the model group, the content of conjugated bile acids in the Olibanum group and vinegar-processed Olibanum group was significantly decreased(P<0.01), the mRNA and protein expressions of FXR, FGF15 and FGFR4 were significantly elevated(P<0.05, P<0.01), and vinegar-processed Olibanum exhibited superior effects than Olibanum. In cellular experiments, a significant decrease in mRNA expression of FXR and FGF19 was observed in the conjugated bile acid group when compared with the blank group(P<0.01). Compared with the conjugated bile acid group, the mRNA expressions of FXR and FGF19 were significantly higher in the Olibanum, vinegar-processed Olibanum and ADHBA groups(P<0.05, P<0.01), and the effect of vinegar-processed Olibanum was more favorable. ConclusionVinegar-processed Olibanum may enhance the ameliorating effect on UC by enhancing the down-regulation of conjugated bile acids in the colon and the up-regulation of FXR-FGF15/19-FGFR4 "gut-liver" crosstalk pathway, and ADHBA may be the main material basis for the synergy.
10.A multicenter phase Ⅲ clinical study of human prothrombin complex concentrate in treatment of hemophilia B
Wei ZHANG ; Yirun LIU ; Yan WU ; Xuanlin ZHONG ; Qingshuang SONG ; Shitao CHEN ; Xueyun WANG ; Caiping GUO ; Zhan ZHANG ; Yunjia ZHANG
Chinese Journal of Blood Transfusion 2022;35(9):915-919
【Objective】 To evaluate the clinical efficacy and safety of one kind of human prothrombin complex concentrate in treatment of patients with hemophilia B. 【Methods】 The clinical data of 36 patients with hemophilia B treated with human prothrombin complex concentrate produced by Shenzhen Weiguang Biological Products Co. Ltd. from May 2018 to April 2019 were retrospectively analyzed, and its clinical efficacy and safety were analyzed. 【Results】 A total of 35 subjects entered the full analysis set (FAS)and safety set (SS), 33 subjects entered the per protocol Set (PPS). Thirty minutes after the first infusion of FAS subjects, the activity of coagulation factor Ⅸ increased from (3.93±0.975) IU/dL to (25.61±9.337) IU/dL, and the infusion efficiency was (96.43±22.007)%. The increased value of coagulation factor Ⅱ activity was (73.25±14.874) IU/dL. The activity of coagulation factor Ⅶ was (42.79±16.847) IU/dL. The increased value of coagulation factor Ⅹ activity was (65.29±17.042) IU/dL. The increased value of coagulation factor Ⅸ activity was (21.68±9.434%) IU/dL. Twenty-four hours after the first infusion of FAS subjects, the improvement of bleeding symptoms and signs was excellent in 21 cases (60%), improved in 14 cases (40.0%), and the effective rate was 100%. The incidence of adverse reactions was 2.9%(1/35), and there was no antibody to human coagulation factor Ⅸ and new virus infection. 【Conclusion】 Infusion of human prothrombin complex concentrate produced by Shenzhen Weiguang Biological Products Co. Ltd. in the treatment of hemophilia B has significant clinical efficacy and good safety.