1.Extraction of Active Compound in Curcuma Phaeocaulis Valeton by Supercritical Fluid Extraction and Trapping with Silica Gel Column
Shulian CHEN ; Jing YOU ; Guojun WANG
Chinese Journal of Analytical Chemistry 2001;29(6):664-666
The essential oil of curcuma phaeocaulis valeto n can be extracted by supercritical fluid CO2 modified with ethanol. The recov ery of active compound, curdione, could be improved when supercritical fluid ext raction with trapping by silica gel column was used and then eluted the analytes with hexane, ether and ethanol. The relative recovery of curdione increased fro m 10.7% to 34.7%.
2.Role of CXCR4 expression in progress and prognosis of breast cancer
Hongwu CHEN ; Guoying WU ; Guojun ZHANG
Journal of International Oncology 2011;38(1):36-38
CXCR4, a receptor for stromal cell-derived factor-1 (SDF-1, also know as CXCL12) and a member of the chemokine receptor family, has been shown to play an important role in the proliferation, migration, invasion and metastasis of breast cancer cells. CXCR4 expression is an early event in breast cancer development and may serve as a biomarker for early detection and diagnosis of breast cancer. Expression of CXCR4 has been demonstrated to correlate with metastasis and poor prognosis in breast cancer. Inhibition of CXCR4 expression may be a potential target of breast cancer treatment.
3.Establishment of Mathematic Model of the Correlativity of HCMV Infection Between Mothers and Their Newborns
Guojun WU ; Shuzheng CHEN ; Gan DAI
Journal of Chinese Physician 2000;0(12):-
Objective To establish a mathematic model of correlativity of human cytomegalovirus(HCMV) infection between mothers and their newborns.Method The plasma HCMV-IgM content,HCMV-DNA levels in plasm and peripheral blood mononuclear cells (PBMCs) were assayed with ELISA and PCR technique.Results There was a evident correlation between the two generation,and there was largest correlativity between HCMV-DNA level of mother's plasma and HCMV congenital infection rate,its spearman rank correlative coefficient was 0 7018(P
4.Application of calcitonin for the treatment of unstable intertrochanteric fractures in elderly patients
Bao CHEN ; Guojun CHEN ; Suiliang GONG ; Chenglong HUANG ; Shunwu FAN
Chinese Journal of Orthopaedics 2014;34(1):24-28
Objective To compare the clinical outcomes between proximal femoral nail antirotation (PFNA) combined with salmon calcitonin and PFNA only for the treatment of unstable intertrochanteric fractures in elderly patients.Methods From January 2009 to December 2011,120 elderly patients with intertrochanteric fracture were randomly divided into two groups:calcitonin group and control group.Patients in calcitonin group were treated with PFNA combined with salmon calcitonin,while patients in control group were treated with PFNA only.According to Evans-Jensen classification,60 patients in calcitonin group (28 males and 32 females,with an average age of 75.1 years) were divided into 20 cases of type Ⅱ A,32 cases of type Ⅱ B and 8 type Ⅲ.Sixty patients in control group (27 males and 33 females,with an average age of 74.9 years) were divided into 22 cases of type Ⅱ A,32 cases of type Ⅱ B and 6 type Ⅲ.Bone healing was assessed with X-ray and bone mineral density (BMD) was measured by dual energy X-ray absorptiometry.Harris hip and SF-12 score,complications,adverse effect of salmon calcitonin and subsequent fragility fractures were evaluated postoperatively.Results One-hundred and thirteen patients were followed up for at least 2 years.In 6 months after surgery,there were 4 cases of delayed healing in control group.However,all fractures were healed in 12 months after surgery.No significant difference was found between the two groups in BMD preoperatively.The changes in BMD were significantly different between the two groups in 6 months,1 year and 2 years after surgery.No significant difference was found between the two groups in Harris hip and 1-year SF-12 score while the 2-year SF-12 score was significantly different between the two groups.There was 1 patient in calcitonin group who suffered from subsequent fragility fracture in 3 months after surgery,while there was 6 patients in control group during 13 to 23 months postoperatively.Conclusion PFNA combined with salmon calcitonin achieves good effect for the treatment of unstable intertrochanteric fractures in elderly patients.
5.Expression, purification and characterization of a thermostable lactate dehydrogenase from Thermotoga maritima.
Guojun QIAN ; Caiping CHEN ; Ruying ZHAI ; Weilan SHAO ; Yanzhen MEI
Chinese Journal of Biotechnology 2014;30(4):545-553
The gene encoding thermostable lactate dehydrogenase (Tm-LDH) was cloned into the plasmid pHsh from Thermotoga maritima, and expressed in Escherichia coli JM 109. The recombinant protein was purified to homogeneity by a simple step, heat treatment. The recombinant enzyme had a molecular mass of 33 kDa. The optimal temperature and pH of Tm-LDH were observed 95 degrees C and 7.0. The purified enzyme had a half-life of 2 h at 90 degrees C, and exhibited better stability over a pH range from 5.5 to 8.0. The K(m) and V(max) values were 1.7 mmol/L, 3.8 x 10(4) U/mg of protein for pyruvate, and 7.2 mmol/L and 1.1 x 10(5) U/mg for NADH, respectively. The expression of Tm-LDH in T7 system could not obtain high efficiency, but it has been soluble over-expression in pHsh system and reached 340 mg/L. The superior stability and productivity of Tm-LDH will lay the foundation of its industrial-scale fermentation and application in the NAD regeneration.
Cloning, Molecular
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Enzyme Stability
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Escherichia coli
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metabolism
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L-Lactate Dehydrogenase
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biosynthesis
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Molecular Weight
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Recombinant Proteins
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biosynthesis
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Temperature
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Thermotoga maritima
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enzymology
6.Glycosyl isomerization based on the biosynthesis of natural-product sugar from microorganism.
Wan SUN ; Haifeng LI ; Jing CHEN ; Guojun WANG ; Zhaoyong YANG
Acta Pharmaceutica Sinica 2013;48(2):179-86
Glycosylation, one of the most common and important reactions in biological systems, results in diverse functions and is often found in biologically active small-molecule natural products produced by microorganisms. Furthermore, sugar moieties are generally critical for their activities. Alternating the sugar structures thus provides the potentials for enhancing the biological activities of natural products, which evokes researchers to study the sugar biosynthetic machinery and its application in the modification of sugar moieties with an aim of generating unnaturally glycosylated natural product drugs with better activities. This review will briefly outline current studies on sugar biosynthesis and glycosyltransferase, with a few selected experiments designed to alter natural-product sugar structures.
7.Isolation and culture of human bone marrow mesenchymal stem cells without lysing the erythrocytes
Guojun CHEN ; Yali WANG ; Feng FANG ; Houqi LIU
Clinical Medicine of China 2012;28(4):344-346
Objective To observe the effects of red blood cell lysis buffer on the isolation and culture of human bone marrow mesenchymal stem cells (hMSCs) in vitro.Methods Twenty-two bone marrow samples were randomly divided into 2 groups,including 11 samples of crushed red blood cell prepared with Tris-NH4Cl red blood cell lysis agent and 11 samples prepared without red blood cell lysis agent.The intervals from primary generation to 1,2 and 3 passages and the time of expansion to l07 cells were compared between the two groups.Results The times for P1,P2 and P3 passage was ( 9.3 ± 4.9 ) days vs.( 7.2 ± 1.0 ) days,( 14.4 ±4.7) days vs.( 14.5 ± 3.5 ) days,and ( 18.5 ± 5.0 ) days vs,( 20.1 ± 4.4 ) days,respectively,in crushed red blood cell group and non-crushed red blood cell group.The differences were not significant ( t =1.39,t =0.06,t =0.80,P > O.05 ).The time for expansion to 107 cells in two groups was not significantly different ( t =0.80,P >0.05).Conclusion Tris-NH4Cl agent in red blood cell lysis has no significant effects on hMSCs isolation,culture and cell proliferation,which indicates erythrocyte lysing may be not an independent step for hMSCs isolation and expansion in vitro.
8.Effect of hydrogen-rich saline on allergic contact dermatitis in mice
Chenyang MENG ; Hongguang CHEN ; Xiaoyin MENG ; Weibin XING ; Guojun FU
Chinese Journal of Dermatology 2013;(4):275-278
Objective To evaluate the therapeutic effect of hydrogen-rich saline on allergic contact dermatitis (ACD) in mice and to explore its underlying mechanisms.Methods Forty mice were equally divided into 4 groups:control group,control treatment group,ACD group and ACD treatment group.ACD was induced by repetitive topical application of dinitrofluorobene (DNFB) to the left ear of mice on day 1,2 and 5.Hydrogen-rich saline was intraperitoneally given to the mice in the ACD treatment group at a dose of 5 ml/kg per day from day 1 to 5.On day 6,the mice were sacrificed,ear tissue was removed from them and subjected to measurement of thickness and weight,detection of tumor necrosis factor (TNF)-α,interleukin (IL)-6,IL-17 and interferon (IFN)-γ expression by enzyme linked immunosorbent assay,as well as numeration of inflammatory cells in lesions after hematoxylin-eosin (HE) staining.Data were processed by SPSS 18.0 software,and statistical analysis was carried out by one-way analysis of variance and least significant difference (LSD) procedure.Results Compared with the control group,the ACD group showed a significant increase in lesion score (7.33 ± 1.53 vs.0,P < 0.05),differences in the thickness ((0.73 ± 0.15) mm vs.(0.13 ± 0.05) mm,P < 0.05) and swelling degree (expressed as tissue weight:(18.67 ± 3.05) mg vs.(3.33 ± 1.52) mg,P < 0.05) between the left and right ear,expressions of TNF-α ((1475.52 ± 233.81) pg/mg vs.(239.01 ± 52.39) pg/mg,P< 0.05),IL-6 ((184.65 ± 78.39) pg/mg vs.(42.28 ± 17.64) pg/mg,P< 0.05),IL-17 ((628.56 ± 201.44) pg/mg vs.(127.58 ± 50.28) pg/mg,P< 0.05) and IFN-γ ((197.72 ± 37.81) pg/mg vs.(24.57 ± 8.31) pg/mg,P < 0.05),and the number of inflammatory cells per square millimetre in the left ear tissue (752.00 ± 166.06 vs.127.33 ± 77.18,P < 0.05).However,hydrogen-rich saline treatment induced a statistical decrease in all of these parameters in the ACD treatment group compared with the ACD group,including lesion score (3.33 ± 0.58,P < 0.05),difference in thickness ((0.46 ± 0.11) mm,P < 0.05) and swelling degree ((11.00 ± 2.64) mg,P < 0.05),expressions of TNF-α ((817.72 ± 101.13) pg/mg,P< 0.05),IL-6 ((95.86 ± 36.65) pg/mg,P< 0.05),IL-17 ((373.38 ± 126.74) pg/mg,P< 0.05),IFN-γ ((63.31± 17.38) pg/mg,P < 0.05) and the number of inflammatory cells per square millimetre (384.00 ± 97.35,P <0.05).Conclusion Hydrogen may inhibit the release of inflammatory factors in ACD.
9.Silencing IDH-2 gene by siRNA-IDH-2 inhibits human small cell lung car-cinoma growth
Jianhong LU ; Guojun CHEN ; Changlin DONG ; Shaowen GUO ; Yijun JIN
Chinese Journal of Pathophysiology 2014;(8):1384-1387
[ABSTRACT]AIM:Toinvestigatetheeffectofsilencingisocitratedehydrogenase2(IDH-2)genebysmallinter-fering RNA (siRNA) on the biological characteristics of human small cell lung cancer cell line NCI -H446.METHODS:IDH-2 expression was knocked down in human small cell lung cancer cell line NCI -H446 by siRNA-IDH-2.The expression level of IDH-2 was determined by real-time PCR and Western blotting .The cell proliferation was measured by CCK-8 as-say , the protein expression of MAPK p 42 was detected by Western blotting , and the cell cycle was analyzed by flow cytome-try.The migration was observed using Transwell cell migration system .BALB/c nude mice were subcutaneously injected on the back with NCI-H446 cells transfected with siRNA-IDH-2/negative control siRNA or non-transfected cells to study the tumor growth .RESULTS:siRNA-IDH-2 remarkably down-regulated the expression of IDH-2 and MAPK p42 in the NCI-H446 cells.siRNA-IDH-2 inhibited both the proliferation and migration abilities of NCI-H446 cells, and the cell cycle was arrested in S phase as compared with negative control group .Additionally, the volume of xenograft tumors in siRNA-IDH-2 group was significantly decreased as compared with control group .CONCLUSION:siRNA-IDH-2 down-regulates the expres-sion of IDH-2 in NCI-H446 cells, reduces the cell migration efficiency and inhibits the tumor growth in vitro and in vivo.
10.Quantitative site of the related structures for approach of blocking the facial nerve
Fu WANG ; Congru QI ; Guojun YANG ; Zhihong CHEN
Chinese Journal of Tissue Engineering Research 2005;9(17):194-195
BACKGROUND: The blocking treatment can improve the clinic symptom of facial spasm. But the site, depth and the angle of puncturing point are very difficult to be defined, which will seriously infect the curative effect.OBJECTIVE: To study the applied anatomy of the related structures of facial muscle for blocking the facial nerve, and to provide anatomical bases for accurate puncturing point and preventing complications.DESIGN: An observation study based on cadavers and making the mimic puncture and measuring the correlative structures with anatomical method.SETTING: Department of anatomy in a medical college.PARTICIPANTS: Totally 28 adult male cadavers(56 sides) were used and the correlative index were measured.METHODS: The blocking point was located at the convergent point of the interior edge of cartilage of the external acoustic canal, the anterior fringe of mastoid processes and the posterior fringe of mandible ramus, the needle must be thrust vertically to the median sagittal plane. The puncturing needle stopped until it was barred. A longitudinal incision was made from the puncturing point to mandibula angle, and skin, superficial fascia were cut separately. Then carotid superficial fascia was cut from the posterior fringe of parotidean. The facial nerve trunk and the posterior auricular artery were preserved in site. A blunt isolation was made along its incision. Jugular glomus was appeared. Then the correlative index were measured and dealt with statistics method.MAIN OUTCOME MEASURES: The puncturing point, angle and depth,the distance between facial nerve trunk and puncturing point, the position relationship between facial nerve and puncturing needle and the distance relationship between facial nerve and principal structures adjacent to it.RESULTS: The blocking point was located at the convergent point of the interior edge of the external acoustic canal, the anterior fringe of mastoid processes and the posterior fringe of mandible ramus, the needle must be thrust vertically to the median sagittal plane. On the left side, the puncturing depth was (19.91 ± 0.09) mm, On the right side, the puncturing depth was(19.28±0. 10) mm.CONCLUSION: The experimental study could provide accurate puncturing point, angle and depth for blocking the facial nerve and bring the blocking treatment into full play.