1.Purification of Recombinant Fusion Protein Staphylokinase-Hirudin Expressed by Escherichia coli and Analysis of its Dimer
Gen-Shen ZHONG ; Ai-Ping YU ; Ji-De JIN ; Zhong-Hua JIANG ; Zu-Ze WU ;
China Biotechnology 2006;0(02):-
The recombinant fusion protein staphylokinase-hirudin(rSFH) was purified from the high density-fermented engineered E.coli by means of ion-exchange chromatography (IEC) and gel filtration (GF). The purity of rSFH reached to more than 98% determined by RP-HPLC and SDS-PAGE, and the yield was up to 0.7g per liter of fermentation broth. The analysis of homologous dimmer of rSFH appeared during the purification and calculation of the surface hydrophobic area had been carried out by means of hydrophobic chromatography and MALD-TOF. The influence of sodium chloride and temperature on the behavior of rSFH reversible dimerization was analyzed by high performance sized- exclusive chromatography(HPSEC). It is concluded that the hydrophobic interaction played an important role in the reversible dimerization of rSFH.
2.In vitro Antiviral activity of a berberine derivant HB-13 against herpes simplex virus
Jian-Bing WU ; Xin-Yu LI ; Lin LIN ; Pan-Gen CUI ; Jia-Run ZHENG ;
Chinese Journal of Dermatology 2003;0(11):-
Objective To evaluate the in vitro antiviral activity of HB-13,a compound derivant from berberine and its prodrug berberine,against herpes simplex virus type 1 (HSV-1) and type 2 (HSV-2). Methods Vero cells were cultured in vitro and infected with HSV.Then,various concentrations of HB-13, berberine,and aciclovir were used to treat these infected cells.The cytopathic effect was observed to deter- mine the antiviral effects and cytotoxicity of HB-13 and berberine.Results For HB-13,berberine and acy- clovir,the half toxicity concentration (TC_(50)) to Vero cells was 31.99,380 and more than 800?g/mL, respectively;the average half inhibitory concentration (IC_(50)) against HSV-1 was 1.328,more than 100,and 0.443?g/mL,respectively,the treatment index (TI) against HSV-1 was 24.09,less than 3.80,and more than 1805.87,respectively;the IC_(50) against HSV-2 was 1.344,more than 100,and 0.679?g/mL,respectively,the TI against HSV-2 was 23.80,less than 3.80 and more than 1178.20,respectively.Conclusion HB-13 possesses marked antiviral activity against HSV-1 and HSV-2 in vitro,while berberine does not.
3.Analysis of epidemiological features of endemic arsenism in Huhhot of Inner Mongolia in 2004-2006
Xiu-fen, LIANG ; Yong-qiang, WANG ; Yuan, SONG ; Gang, WANG ; Yu-gen, WU
Chinese Journal of Endemiology 2010;29(1):66-69
Objective To study the epidemiological features of the endemic arsenism in Hohhot of Inner Mongolia in 2004-2006 in order to get scientific evidence for the control and prevention. Methods The historical data in Huhhot revealed that 51 arsenic villages in 3 counties(Tumotezuo,Tuoketuo and Helingeer) were investigated. When arsenic content in drinking water was 0.05-0.15 mg/L,the situation of endemic arsenism was investigated in the households having high arsenic contents of water. When arsenic content in drinking water was greater than 0.15 mg/L,the survey was conducted to investigate arsenic patients with census. Census rate was not lower than 90%. Patients of endemic amenism were diagnosed by The Standard of Diagnosis for Endemic Arsenism (WS/T 211-2001). Results A total 28 083 people were examined,3978 were determined to be patients with arsenism,including 1722 being suspicious, 1897 mild,329 secondary and 30 severe. The total incidence of arsenism was 14.17% (3978/28 083),while the incidence of suspicious,mild,secondary and severe disease was 6.13% (1722/28 083) ,6.75% (1897/28 083),1.17% (329/28 083) and 0.11% (30/28 083),respectively. Suspicious and mild patients were most frequently seen. The incidence of dyspigmentation,pigmentation,ketatoses,both dyspigmentation and pigmentation and all of skin disorders was 8.65% (2428/28 083),3.19% (895/28 083),6.48% (1821/28 083),2.13%(598/28 083) and 0.94%(265/28 083),respectively. The most seen skin disorders were dyspigmentation and ketatoses. The disease mostly occurred in those aged over 40 years,the incidence was 89.52% (3561/3978). The incidence of male [15.08%(2209/14 644)]was more than that of female[13.16%(1769/13 439)].Conclusions Current condition of arsenism in Hohhot is mild,arsenic disease surveillance should keep going. Prevention and control needs to be reinforced in the future.
4.Neurogenesis in the adult brain and depression
Liu QIONG ; Yu JIN ; Wu GEN-CHENG
Neuroscience Bulletin 2005;21(6):436-440
Neurogenesis can be observed in the adult nervous system of many mammals. It is a popular topic about the regulation and functional reconstruction of adult neurogenesis. As one of the most active areas where neurogenesis occurs, hippocampus is most sensitive to stressful stimulations. Moreover, hippocampus plays a critical role in emotion controlling and may be the basis of research of depression. In addition, many regulative factors of neurogenesis may be involved in the etiology and therapy of depression. At present, the relationship between adult brain neurogenesis and depression should be a new focus for the research of depression.
5.Effects of Fengshi Bitong Prescription on Cytokines in Serum and Synovial Fas/FasL mRNA Expression in Rats with Collagen-Induced Arthritis
Wei LIANG ; Qun ZHOU ; min Jia SHEN ; gen Gen WU ; ling Li ZHANG ; Ye CHEN ; yu Liang YAO
Chinese Journal of Information on Traditional Chinese Medicine 2017;24(12):52-55
Objective To observe the effects of Fengshi Bitong Prescription (FSBT) on cytokines in serum and Fas/FasL mRNA expression system in synovium; To discuss relevant mechanism of action. Methods Rats were given foot intradermal injection of bovine type Ⅱ collagen emulsion to establish collagen-induced arthritis (CIA) model. After successful modeling, they were randomly divided into 6 groups: normal control group, CIA model group, tripterygium glycosides group, FSBT high-, medium-, and low-dose groups. Each medication group was given relevant medicine for gavage for 14 d. The weight and arthritis scores of CIA rats were observed. Serum levels of IL-1β, IL-15, and TGF-β1 were detected by ELISA. Expressions of Fas and FasL mRNA in synovium of CIA rats were detected by real-time PCR. Results Compared with model group, the foot arthritis score and serum levels of IL-15 in FSBT high- and medium-dose groups significantly decreased (P<0.01), and the serum level of TGF-β1 significantly increased (P<0.05). Expression of Fas significantly decreased and FasL significantly increased in FSBT high-dose group (P<0.05). Conclusion FSBT may have certain immune regulation effects on rheumatoid arthritis. Its mechanism may be in regulating Fas/FasL apoptosis system, thereby inducing apoptosis of synovial cells and inhibiting synovial hyperplasia.
6.Effect of vitamin E on the lag time of low-density lipoprotein oxidation in vitro in patients with cerebral infarction
Wei-Ping GUAN ; Pei-Gen KUANG ; Sheng-Yuan YU ; Wei-Ping WU ; Hirofumi METOKI
Chinese Journal of Neuromedicine 2008;7(12):1249-1251
Objective To investigate the effect of vitamin E on the lag time of serum low-density lipoprotein (LDL) oxidation in vitro in patients with acute cerebral infarction (ACI). Methods Twenty-two patients with ACI received vitamin E treatment at the dose of 300 mg/day, and another 22 ACI patients matched for age, gender ratio, blood pressure, blood lipid level, location and degree of the infarction served as the control group in which no vitamin E was given. Serum samples were collected from the patients within 24 h and at 2 weeks after the onset to test the lag time of LDL oxidation in vitro and measure the levels of vitamin E, total cholesterol, triglyceride and free cholesterol. Results At two weeks after the onset of the infarction, the patients receiving vitamin E treatment had significantly prolonged lag time of LDL oxidation in comparison with that of the control patients (71.84± 9.75 vs 60.95±9.33 min, P<0.05), and showed also significantly higher vitamin E level in the serum Vitamin E and stroke index. The two groups of patients showed no significant differences in total cholesterol (5.08cholesterol (1.62oxidation in vitro is prolonged in ACI patients with vitamin E treatment, suggesting that vitamin E can increase the resistance of the patients against anti-oxidative stress.
7.Estimation of oxidative stress in patients with Parkinson's disease
Wei-Ping GUAN ; Pei-Gen KUANG ; Sheng-Yuan YU ; Wei-Ping WU ; Hirofumi METOKI
Chinese Journal of Neuromedicine 2009;8(1):54-56
Objective To measure the changes in urinary 8-iso-prostaglandin-F2α (8-iso-PGF2α) and oxidized low-density lipoprotein (ox-LDL) lag time in patients with Parkinson's disease (PD). Methods A case-control study was performed involving 31 male patients with PD (mean age of 59.7 years) diagnosed according to the Calne criteria and 31 age-matched healthy male subjects with comparable status of smoking and life style. For each subject, urinary 8-iso-PGF2α was measured quantitatively using enzyme-linked immunosorbent assay, and LDL oxidizability was measured by determining LDL oxidation lag time in conjugated diene product at 234 nm using Cu-stimulated oxidation. Results Compared to the levels in the control subjects, the PD patients showed significantly increased urinary 8-iso-PGF2α (81.1±1.6 vs 46.9±1.1 ng/mmol creatinine, P<0.05) and significantly reduced LDL oxidation lag time (63.5±6.0 vs 84.4±8.8 min, P<0.05). Conclusion Increased 8-iso-PGF2α and decreased anti-oxidant ability are implicated in the pathogenesis of PD, suggesting the value of appropriate antioxidant therapy in controlling the progression of PD.
8.Melatonin attenuates the intensity of β-endorphin immunoreactivity in the arcuate nucleus of rat hypothalamus
Chang-Xi YU ; Gen-Cheng WU ; Shao-Fen XU ; Chong-Hong CHEN
Acta Physiologica Sinica 2000;52(3):263-266
The present study was undertaken to observe the effect of exogenously administered melatonin on the intensity of β-endorphin (β-Ep) immunoreactivity of the neuron in the arcuate nucleus of rat hypothalamus with an aim to explore the possible mechanisms of the analgesic effect of melatonin. The experimental rats were divided into two groups, one injected intraperitoneally with melatonin (110 mg/kg) and the other with only a vehicle. One hour after injection, the brain was processed for coronal sections, which were stained with immunohistochemical ABC technique. The integral optical density (IOD) and mean optical density (OD) of the stained sections were measured with a computer-assisted image-processing and analytical system. β-Ep immunoreactivity was much decreased in the sections treated with melatonin and the IOD and OD were also decreased significantly (P<0.01; P<0.05). The above results suggest that melatonin may result in a decrease of β-Ep content in the arcuate nucleus, as a result of increased β-Ep release induced by administration of melatonin. It is likely that the analgesic effect of melatonin may be in part mediated by the release of β-endorphin from the arcuate nucleus.
9.Perineal raphe with special reference to its extension to the anus: a histological study using human fetuses.
Zhe Wu JIN ; Yu JIN ; Xiang Wu LI ; Gen MURAKAMI ; José Francisco RODRÍGUEZ-VÁZQUEZ ; Joerg WILTING
Anatomy & Cell Biology 2016;49(2):116-124
The raphe of the human penis and scrotum is considered to develop secondarily after disappearance of the initial midline seam by fusion of the bilateral genital folds. However, the fetal development was still obscure. We examined histological sections of 30 fetuses (17 males and 13 females) at 10-15 weeks. In male fetuses, the scrotum was not yet clearly identified because of no descent of testis. The perineal raphe was thin and wavy at 10 weeks, and it was continuous with and took a direction same as the inferior wall of the closed penile urethra after physiological hypospadias. Depending on growth of the bulbospongiosus muscle and corpus spongiosus penis, the midline intermuscular septum obtained a connection to the subcutaneous wavy raphe and made the latter thick and straight at 12-15 weeks. Notably, the perineal raphe extended posteriorly to attach to the external anal sphincter. In female fetuses, an epithelial fusion occurred along a short distance at the posterior end of the vestibule. However, in front of the external anal sphincter, a large midline mesenchymal tissue from the urorectal septum did not contain a raphe-like structure. Moreover, since the bilateral bulbospongiosus muscles were separated widely by the vestibule, they did not provide a midline septum. Fetal development of the perineal raphe was accelerated by reinforcement from the muscular septum. In contrast, without such a muscular support, the female raphe could not maintain its growth even if the seed appeared at the posterior end of the vestibule.
Anal Canal*
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Female
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Fetal Development
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Fetus*
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Humans*
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Hypospadias
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Male
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Muscles
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Penis
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Scrotum
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Testis
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Urethra
10.Research on HBV DNA inhibition of plasmid acute infection mouse with betulinic acid.
Bing QIAO ; Yue-Qiu GAO ; Man LI ; Shao-Fei WU ; Chao ZHENG ; Shu-Gen JIN ; Hui-Chun WU ; Zhuo YU ; Xue-Hua SUN
China Journal of Chinese Materia Medica 2014;39(6):1097-1100
Betulinic acid is a naturally occurring pentacyclic triterpenoid, which has antiretroviral, antimalarial, and anti-inflammatory properties. The purpose of this study is to investigate the HBV DNA replication inhibition in the mouse model with betulinic acid. Hydrodynamic injection method via the tail vein with the Paywl. 3 plasmid was used to establish the animal mode (n = 15), and the mice were randomly divided into the PBS control group (n = 5), Betulinic acid treatment group (n = 5) and lamivudine control group (n = 5). The day after successful modeling , the mice would have taken Betulinic acid (100 mg x kg(-1)), lamivudine (50 mg x kg(-1)), PBS drugs orally, once daily for 7 days, blood samples were acquired from the orbital venous blood at 3, 5, 7 days after the administering, HBsAg and HBeAg in serum concentration were measured by ELISA and the mice were sacrificed after 7 days, HBV DNA southern detections were used with part of mice livers. The results showed that betulinic acid significantly inhibited the expression of HbsAg in the mice model at the fifth day compared with the control group, and there was no significant differences between the effects of lamivudine and the PBS control group; both the betulinic acid and lamivudine groups had no significant inhibition for the HBeAg expression; the HBV DNA expressions of the liver tissue from the betulinic acid and lamivudine groups were inhibited compared with the control group. Taken together, these results reveal betulinic acid can inhibit the HBsAg expression and replication of the liver HBV DNA in the mouse model.
Acute Disease
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Animals
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Antiviral Agents
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pharmacology
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DNA Replication
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drug effects
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DNA, Viral
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biosynthesis
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Hepatitis B
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blood
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virology
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Hepatitis B Surface Antigens
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blood
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Hepatitis B virus
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drug effects
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genetics
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immunology
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physiology
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Male
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Mice
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Plasmids
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genetics
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Triterpenes
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pharmacology
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Virus Replication
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drug effects