1.The research of C1q/TNF-related proteins
Hao JIA ; Ting WU ; Qin MA ; Jianli ZHAO
Journal of Chinese Physician 2016;(z1):223-225
C1q /TNF-related proteins (CTRPs)belong to the same category of C1q /TNF-related protein family,mainly secreted by adipose tissue and widely expressed in human and rodent.Their function turned out to be metabolic modulation,cardiovascular system protection and suppression of inflammation.But for metabolic modulation,enhancing the synthesis of glycogen,promoting fatty acid uptake and oxidation,in-creasing insulin sensitivity is the major role of CTRPs playing in governing the metabolism.Each CTRP has its own unique tissue expression profile and nonredundant function in regulating sugar and /or fat metabo-lism.In this review we focus on the recent progress about the metabolic function of CTRPs.
2.Initial study on the role of bacterial flagellin and CD98 in ulcerative colitis
Huijie HAO ; Fumin XUE ; Jianli ZHANG ; Chuan ZHOU ; Linjing LI ; Baisui FENG
Chinese Journal of Digestion 2012;32(9):625-628
Objective To investigate the role of bacterial flagellin and CD98 in ulcerative colitis (UC).Methods A total of 60 first episode patients with active UC were recruited,including 30 mild and 30 moderate to severe UC cases.The serum of 30 healthy volunteers and normal intestinal tissues surgically removed from 15 colon cancer patients (more than 5 cm away from surgical margins) were collected as control.The content of bacterial flagellin antibodies in peripheral blood were measured with enzyme-linked immunosorbent assay (ELISA).The expression of CD98 in peripheral blood T lymphocyte was measured by flow cytometry (FACS).The expression of bacterial flagellin protein in intestinal mucosa and CD98 in intestinal epithelial basement membrane was tested by immunohistochemistry (IHC).The comparison between two groups was performed with the SNK-q method,the R×C table x2 test was used to analyze the counted data,and the Spearman correlation was used to analyze the rank materials.Results The peripheral blood concentration of bacteria flagella protein antibody of control group,mild UC group and moderate to severe group showed an upward trend,which was (7.603±2.118) pg/ml,(13.702±3.131) pg/ml and (20.813±3.004) pg/ml respectively,and the differences among groups were statistically significant (F=13.57,P<0.01).The expression percentage of bacteria flagella protein in intestinal mucosa of the three groups also showed an upward trend,which was 3/15,56.67% and 73.33% respectively,and the differences among groups were statistically significant (x2 =11.553,P=0.003).The positive rate of CD98 expression in peripheral blood T lymphocytes of the three groups showed an upward trend,which was (28.42±4.31)%,(32.45±6.71)% and (43.40±5.09) % respectively,and the differences among groups were statistically significant (x2 =10.110,P=0.007).The positive rate of CD98 expression in intestinal epithelial cells of the three groups also showed an upward trend,which was 1/15,36.67 % and 66.67% respectively,and the differences among groups were statistically significant (x2 =5.400,P<0.05).There was positive correlation between the peripheral blood concentration of bacteria flagella protein antibody and the expression of CD98 in peripheral blood T lymphocytes (r=0.548,P<0.05).Conclusion Bacterial flagellin and CD98 may be important factors causing inflammatory reaction activity in UC.
3.Determination of Bacterial Endotoxin in Influenza Split Vaccine by a Dynamic Turbidity Method
Yan ZENG ; Lizi DONG ; Yue CUI ; Yan LAN ; Jianli HAO ; Hongyu YANG
China Pharmacist 2017;20(1):171-174
Objective:To investigate the feasibility of dynamic turbidity method for the detection of bacterial endotoxin content in influenza split vaccine. Methods: According to the bacterial endotoxin detection method described in general rule 1143 in Chinese Pharmacopoeia (2015 edition), the reliability test for standard curves of influenza split vaccine, the interference initial screening test, the interference verification test and the endotoxin content were performed or determined, and the results were compared with those by the gel method for the same batches of vaccine. Results:The results of reliability test for standard curves were accordance with the reg-ulations. In the interference initial screening test, vaccine was diluted by 160 times, 320 times and 640 times, and the recovery was between 50% and 200%, which showed no interference. The results of the interference verification test further proved that vaccine with 640 times dilution had no interference effect on test. The bacterial endotoxin contents of 10 batches of influenza split vaccine deter-mined by the turbidity method were less than the limit value of 20 EU·ml-1 , and the results were the same as those determined by the gel method. Conclusion:It is feasible to detect the content of bacterial endotoxin in influenza split vaccine by the dynamic turbidity method, which is worthy of promoted application.
4.Optimal expression condition of recombinant RAP.
Jie, ZHANG ; Hong, ZHANG ; Hao, BI ; Zhiguo, LIU ; Jianli, GUO ; Shen, QU
Journal of Huazhong University of Science and Technology (Medical Sciences) 2007;27(1):5-8
In order to construct the expression recombinant of human receptor associated protein (RAP), optimize its expression condition and obtain the recombinant protein after expression with high efficiency, two prokaryotic expression vectors-pT7-PL and pET-28a(+) were used to construct the expression recombinant containing RAP cDNA, and the expression efficiency of two kinds of expression E. coli of BL21 strains was compared. The effect of different induction conditions on the expression of recombinant RAP was observed. After recombinant protein was purified with Ni(+) -nitrilotriacetic acid (Ni(+) -NTA) affinity chromatogram, its binding ability with microphage was observed. The results showed that two recombinant plasmids both obtained high expression of RAP. The expression levels of RAP in plasmid pT7-PL-RAP in BL21 (DE3, plysS) strain were significantly higher than in BL21 (DE3) strain. The expression of pT7-PL-RAP in the presence of chloramphenicol was higher than in the absence of chloramphenicol, and most of the inducible expressed RAP was soluble. The RAP which was purified by Ni(+) -NTA resin could strongly bind with the RAW264.7 cells rich in low density lipoprotein receptor (LDLR) family receptors. It was concluded that the expression condition of recombinant RAP was optimized and functional RAP was obtained, which offered a good foundation for the further production of RAP as research tool.
5.Cloning and Expression of Apolipoprotein E3 and Its Variant apoE2 and apoE4
Yiqiang ZONG ; Zhiguo LIU ; Hao BI ; Yanyi YAO ; Jianli GUO ; Shen QU
Journal of Huazhong University of Science and Technology (Medical Sciences) 2006;26(1):1-3,16
In order to obtain three isoforms of apolipoprotein E (apoE), the cDNA encoding apoE3 was obtained by RT-PCR from normal human liver tissue. Site-directed mutagenesis was used to obtain the cDNAs encoding apoE2 and apoE4 isoforms. The 3 cDNAs were subcloned into vector pGEM-3Z and verified by DNA sequencing. The expression recombinant which can express the target protein as a (His) 6-tagged fusion was constructed by subcloning apoE cDNA into vector pT7-PL. The purified proteins were gained by Ni-NTA column. The SDS-PAGE results revealed the 6 His fusion proteins (apoE2, apoE3 and apoE4) were correctly expressed and purified successfully.
6.Optimal Expression Condition of Recombinant RAP
Jie ZHANG ; Hong ZHANG ; Hao BI ; Zhiguo LIU ; Jianli GUO ; Shen QU
Journal of Huazhong University of Science and Technology (Medical Sciences) 2007;27(1):5-8
In order to construct the expression recombinant of human receptor associated protein (RAP), optimize its expression condition and obtain the recombinant protein after expression with high efficiency, two prokaryotic expression vectors-pT7-PL and pET-28a(+) were used to construct the expression recombinant containing RAP cDNA, and the expression efficiency of two kinds of expression E. coli of BL21 strains was compared. The effect of different induction conditions on the expression of recombinant RAP was observed. After recombinant protein was purified with Ni+-nitrilotriacetic acid (Ni+-NTA) affinity chromatogram, its binding ability with microphage was observed. The results showed that two recombinant plasmids both obtained high expression of RAP. The expression levels of RAP in plasmid pT7-PL-RAP in BL21 (DE3, plysS) strain were significantly higher than in BL21 (DE3) strain. The expression of pT7-PL-RAP in the presence of chloramphenicol was higher than in the absence of chloramphenicol, and most of the inducible expressed RAP was soluble. The RAP which was purified by Ni+-NTA resin could strongly bind with the RAW264.7 cells rich in low density lipoprotein receptor (LDLR) family receptors. It was concluded that the expression condition of recombinant RAP was optimized and functional RAP was obtained, which offered a good foundation for the further production of RAP as research tool.
7.Thoracoumbilical flap combined with random abdominal flap in repair of large-area soft tissue defects of calf in children
Hao ZHENG ; Yong LIU ; Lili WANG ; Jianli WANG ; Xuejian GAO
Chinese Journal of Microsurgery 2022;45(5):528-533
Objective:To investigate the clinical effect of thoracoumbilical flap combined with random abdominal flap in repair of large-area soft tissue defects of calf in children.Methods:The clinical data of 16 children with large-area soft tissue defects of calf treated with thoracoumbilical flap combined with abdominal random flap from January 2004 to December 2007 in PLA Trauma Orthopaedic Research Institute, 80th Group Military Hospital of the PLA were retrospectively analysed. There were 7 boys and 9 girls aged 8 to 14(11.3 in average) years old. Six cases were crushed by heavy objects, 6 crushed by wheels, 3 by thermal press and 1 by machine strangulation. After thorough debridement, the wound area ranged from 16.0 cm×9.0 cm to 38.0 cm×15.0 cm. Four cases were treated after 3-10 hours of injury by emergency surgery. Twelve cases received surgeries 0-11 days after hospital admission and wound being stabilised. Doppler ultrasound was used to locate the perforating vessels according to the location, size and shape of the wound. Thoracoumbilical flap combined with abdominal random flap were designed and harvested to repair the wound. The sizes of flaps were 18.0 cm×11.0 cm-40.0 cm×16.0 cm. All patients entered follow-up at the outpatient clinic or through WeChat interviews. The appearance, texture of the flap and limb recovery were checked and recorded.Results:After surgery, all of 16 flaps survived, of which 12 flaps had phase-one healing, 3 flaps had small area of necrosis at the edge, which healed after repeated dressing changes and 1 flap developed vascular comproise, and survived after surgical exploration. The donor sites healed in phase-one. All 16 children had 6 months to16 years of follow-up, with an average of 20.7 months. The colour of the flaps was normal with soft texture. The motor function of calf was satisfactory. According to Punor functional evaluation criteria, 12 cases were in excellent and 4 in good.Conclusion:The thoracoumbilical flap combined with abdominal random flap features a reasonable design, strong blood supply and repair of a large area. It is a reliable method for repairing large area soft tissue defects in the calf of children.
8.Anatomical study and clinical application of cross-donor flaps pedicled with peroneal vessels
Chengqi YU ; Yong LIU ; Jianli WANG ; Hao ZHENG ; Jia GUO ; Zhiqiang SUI ; Jian LI ; Xuetao ZHANG ; Lei WANG
Chinese Journal of Microsurgery 2022;45(1):71-76
Objective:Anatomical study of the cross-donor flap pedicled with the peroneal artery and the discussion of the effect of clinical application, so as to describe a new method for the repair of large-area soft tissue defects in the foot and ankle.Methods:From June 2016 to August 2019, 12 specimens of adult lower limbs were studied. The popliteal arteries were perfused with perchloroethylene-ethyl acetate-lead oxide and red perchloroethylene-ethyl acetate. The origin, number, outer diameter, course and distribution of perforating branches of the peroneal artery were anatomically observed. The source, distribution and anastomosis of the skin nutrient vessels in the posterolateral area of the calf were also studied. Relationship of the blood supply between the peroneal arteries and veins and the nutritional vessels of the sural nerve were observed. In 9 patients, the peroneal artery and vein were designed as the pedicle of cross-donor flap in the repair of large soft tissue defects of foot and ankle. The patients were entered follow-up through outpatient visits and telephone interviews.Results:Among the 12 adult specimens of lower limbs, there were 65 perforating branches from the peroneal artery, 4-7 branches on each side, with an average of (5.41±1.00) branches. The diameter of the penetrating deep fascia was(1.07±0.36) mm. The perforator branches were mostly distributed in 3 sections of 4.0-11.0 cm, 16.0-21.0 cm and 24.0-27.0 cm away from the lateral malleolus, accounting for 48%, 24% and 17% of the total number of perforators, respectively. The outer diameters of the perforator vessels were (0.92±0.26)(0.56-1.68) mm, (1.32±0.38)(0.60-2.14) mm, and (0.98±0.28)(0.62-1.36) mm. The length of the pedicle of the perforator vessels were (3.91±0.96)(2.15-5.78) cm, (5.34±0.50)(4.01-5.85) cm, and (3.31±1.15)(2.16-5.66) cm. The perforating branches in the 3 sections appeared constantly. The diameter of the vessels was≥0.5 mm with an average length of at(4.19±1.16)(2.15-5.85) cm. The vascular network of the flap in the posterolateral area of the calf was mainly composed of subdermal vascular network and deep fascial vascular network. The deep fascia vascular network in the posterolateral area of the calf had 3 obvious longitudinal chains, including the medial sural neurotrophic vascular chain, the small saphenous vein-sural nerve communicating branch vascular chain and the lateral sural neurotrophic vascular chain, which took the nutrient blood supply from the perforating branches of the peroneal artery also formed a longitudinal and transverse anastomosis between the perforating branches of the peroneal artery. In the clinical trials performed on 9 patients, all soft tissue defects of foot and ankle were repaired. The composite tissue flap survived without infection or necrosis. The follow-up was lasted for 12 months to 3 years. The postoperative function and the donor site appearance were good and the patients walked normally. According to the American Orthopaedic Foot and Ankle Association(AOFAS) foot scoring standard, the function of affected feet were evaluated. Five patients were excellent and 4 were good.Conclusion:The cross-donor flaps pedicled with peroneal arteries and veins has sufficient blood supply and a large area. It provides a method for the repair of large-area soft tissue defects in the foot and ankle.
9.Ilizarov bone transport assisted by a 3D printed patient-specific guide plate for treatment of tibial bone defects
Hao ZHENG ; Lili WANG ; Yong LIU ; Jianli WANG ; Xuejian GAO
Chinese Journal of Orthopaedic Trauma 2023;25(7):617-623
Objective:To evaluate the efficacy of 3D printed patients-specific guide plates in assisting Ilizarov bone transport in the treatment of tibial bone defects.Methods:A retrospective study was conducted to analyze the clinical data of 24 patients with tibial bone defects who had been admitted to Institute of Trauma Orthopedics, The 80th Army Group Hospital of PLA from January 2018 to March 2022. There were 9 males and 15 females with an age of (49.8±6.5) years, and 4 upper tibial defects, 5 middle tibial defects, and 15 lower tibial defects. According to the methods of repairing bone defects, the patients were divided into 2 groups: a 3D printing group of 10 cases where a 3D printed patient-specific guide plate was used to assist Ilizarov bone transport in the treatment of tibial bone defects, and a traditional group of 14 cases where Ilizarov bone transport was performed in a traditional manner. The 2 groups were compared in terms of operation time, frequency of intraoperative fluoroscopy, axial angulation of the tibia at postoperation and the last follow-up, external fixation time (EFT) and external fixation index (EFI). At the last follow-up, healing of bone defects was evaluated according to the criteria of The Association for the Study and Application of the Method of Ilizarov (ASAMI), functional outcomes were evaluated according to the Paley criteria, and needle infection was recorded according to the Paley classification for complications.Results:There was no statistically significant difference in the preoperative general data between the 2 groups, indicating comparability ( P>0.05). All patients were followed up for (11.3±2.0) months on average after operation. The 3D printing group had significantly shorter operation time [(19.9±2.6) min] and significantly lower frequency of intraoperative fluoroscopy [(3.0±0.8) times] than the traditional group [(38.1±2.2) min and (8.9±1.3) times] (P<0.05), and had significantly better axial angulation of the tibia at postoperation and the last follow-up than the traditional group ( P<0.05). There was no significant difference in EFT or EFI between the 2 groups ( P>0.05), and the last follow-up revealed no significant difference either in bone healing, functional outcomes, or needle infection between the 2 groups ( P>0.05). Conclusion:In the treatment of tibial bone defects, compared with conventional Ilizarov bone transport, the Ilizarov bone transport assisted by a 3D printed patient-specific guide plate demonstrates advantages of shorter operation time, lower intraoperative fluoroscopy, and higher reduction accuracy.
10.Effects of phlegm-resolving and stasis-removing herbal drugs on PGC-1α mRNA expressions and insulin resistance in NAFLD rats
Jianli GUO ; Shuai LU ; Chenxu LIU ; Yu LIU ; Xue HAN ; Lei HAO ; Yixin ZHANG
International Journal of Traditional Chinese Medicine 2019;41(3):252-257
Objective To explore the mechanism of phlegm-resolving and stasis- removing herbals on NAFLD by observing expressions of PGC1α mRNA and insulin resistance. Methods A total of 60 male SD rats were randomly divided into the normal group, model group, positive medication control group, high-dose, middle-dose and low-dose group. The rats were fed with high-fat forage for 8 weeks. The positive medication control group were gavaged with Dongbao-Gantai liquid (0.9 g/kg/day), the high-dose, middle-dose and low-dose group were gavaged with Xiaotan-Huayu liquid (43.34、32.50、21.67 g/kg/day), and normal group, model group were gavaged with equal volume of distilled water. The drugs were given by 1 ml/100 g and last for 8 weeks. The levels of TC, TG, FFA, ALT, AST, FBG, FINS, and HOMA-IR in serum, and levels of TC, TG, and PGC-1α mRNA and pathological morphological changes in hepatic tissue were observed after 8 weeks. Results The levels of TG (0.55 ± 0.10 mmol/L, 0.58 ± 0.09 mmol/L, 0.67 ± 0.11 mmol/L vs. 1.18 ± 0.15 mmol/L), TC (1.48 ± 0.24 mmol/L, 1.69 ± 0.27 mmol/L, 1.74 ± 0.27 mmol/L vs. 3.29 ± 0.26 mmol/L), FFA (251.08 ± 48.18 μmol/L, 277.53 ± 56.73 μmol/L, 291.82 ± 48.67 μmol/L vs. 432.19 ± 67.83 μmol/L), ALT (29.32 ± 4.17 U/L, 31.26 ± 4.74 U/L, 33.56 ± 5.18 U/L vs. 47.21 ± 8.67 U/L), AST (11.05 ± 2.18 U/L, 12.15 ± 2.67 U/L, 12.96 ± 2.93 U/L vs. 19.43 ± 3.68 U/L), FBG (5.68 ± 1.22 mmol/L, 6.86 ± 1.36 mmol/L, 7.94 ± 1.82 mmol/L vs. 11.88 ± 2.54 mmol/L), FINS (8.48 ± 1.22 mmol/L, 9.55 ± 1.95 mmol/L, 9.96 ± 1.74 mmol/L vs. 12.96 ± 2.67 mmol/L), HOMA-IR (1.91 ± 0.26, 2.91 ± 0.65, 3.52 ± 0.58 vs. 6.89 ± 1.21) in serum of high-dose, middle-dose and low-dose groups were decreased than model group. Levels of FFA (242.19 ± 35.13 μmol/L, 259.78 ± 29.33 μmol/L, 277.62 ± 34.29 μmol/L vs. 436.48 ± 52.15 μmol/L), TG (23.65 ± 3.28 mmol/L, 24.41 ± 3.15 mmol/L, 25.37 ± 3.59 mmol/L vs. 15.98 ± 2.37 mmol/L), TC (7.15 ± 0.82 mmol/L, 8.60 ± 0.95 mmol/L, 8.86 ± 1.04 mmol/L vs. 36.98 ± 4.28 mmol/L) were in hepatic tissue of high-dose, middle-dose and low-dose groups were significantly lower than the model group. The levels of PGC-1α mRNA (1.24 ± 0.06, 1.02 ± 0.07, 0.99 ± 0.08 vs. 0.43 ± 0.06) in hepatic tissue of high-dose, middle-dose and low-dose groups were significantly higher than model group. Conclusions The phlegm-resolving and stasis-removing herbals may improve lipid metabolism by regulating the expression of PGC-1α mRNA, inhibiting gluconeogenesis and liver sugar output, correcting disturbance of lipid metabolism and improving insulin resistance.