1.Screening of interferon?binding proteins by yeast-two hybrid technique
Jian-Hui QU ; Jun CHENG ; Ling-Xia ZHANG ;
Chinese Journal of Infectious Diseases 1997;0(04):-
Objective To screen proteins binding with interferon?(IFN?)from human hepatic cDNA libraty by yeast-two hybrid technique.Methods The IFN?gene was amplified by polymerase chain reaction(PCR)and constructed into pGBKT7 vector as the bait plasmid in yeast-two hybrid system3,pGBKT7-IFN?was then transfected into yeast AH109.The transfected yeast were mated with yeast Y187 containing liver cDNA library plasmid in 2?YPDA medium.Diploid yeast was plated on synthetic dropout nutrient medium(SD/-Trp Leu-His-Ade)and synthetic dropout nutrient medium(SD/-Trp-Leu-His-Ade)containing X-?-gal for selecting.After plasmid extracting and en- zyme cutting analysis,the blue colonies were performed sequence analysis,the results were analyzed by bioinformatics.Results IFN?gene was successfully cloned and expressed in yeast cells.Thirty- four positive colonies were obtained using yeast-two hybrid technique.After sequence analysis,eight clones were found may have a binding effect with IFN protein.Conclusions IFN?genes was success- ful cloned and eight proteins that could bind with IFN?protein were also screened.
2.Acute myocarditis misdiagnosed as "acute cholecystitis".
Ying CHEN ; Wei-qiang ZHENG ; Jian-ping ZHONG ; Jian-jun WANG ; Miao-xia HE
Chinese Journal of Pathology 2010;39(2):114-116
Acute Disease
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Adult
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Cholecystitis, Acute
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diagnosis
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pathology
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Diagnostic Errors
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Female
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Humans
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Myocarditis
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diagnosis
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pathology
3.Chemical constituents from bark of Juglans mandshurica
Jin-Lan WANG ; Shu-Xia ZHANG ; Tie-Jun LI ; Wei-Qi ZHANG ; Jian-Jun WANG ; Shu-Jun ZHANG ;
Chinese Traditional and Herbal Drugs 1994;0(04):-
Objective To study the chemical constituents from the bark of Juglans mandshurica. Methods The compounds were isolated and purified by column chromatography on silica gel,HPLC,and recrystallization.Their structures were elucidated by the physicochemical and spectroscopic evidences. Results Fifteen compounds were identified as:4,8-dihydroxynaphthalenyl-O-?D-(6′-acetoxyl)gluco- pyranoside(Ⅰ),dihydrokaempferol(Ⅱ),juglone(Ⅲ),daucosterol(Ⅳ),kaempferol(Ⅴ),4,8-dihy- droxynaphthalenyl-1-O-?-D-[6′-O-(3″,5″-dimethoxy-4″-hydroxybenzoyl)] glucopyranoside(Ⅵ), kaempferol-3-O-?-L-rhamnoside(Ⅶ),3,3′-dimethoxylellagic acid(Ⅷ),naringenin(Ⅸ),quercetin (Ⅹ),reginolone(Ⅺ),quercetin-3-O-?-L-rhamnoside(Ⅻ),naringenin-7-O-?-D-glucoside(ⅩⅢ),4,8- dihydroxynaphthalenyl-1-O-?-glucoside(ⅩⅣ),4,5,8-trihydroxy-?-tetralone-5-O-?-D-[6′-O-(4″-hy- droxy-3″,5″-dimethoxy-benzoyl)] glucoside(ⅩⅤ).Conclusion CompoundⅠ(juglamanol)is a new compound.CompoundsⅡ,Ⅶ—Ⅸ,Ⅻ,andⅩⅢare isolated from plants of Carya Nutt.for the first time.
4.Evaluation and clinical significance of HBV large protein (LHBs) in diagnosis of hepatitis B.
Yong-Jian CHEN ; Yong-Lie ZHOU ; Jun XIA ; Wei-Wei XIA
Chinese Journal of Experimental and Clinical Virology 2007;21(3):241-243
OBJECTIVETo explore the significance of HBV large protein (LHBs) in diagnosis of hepatitis B, we detected the LHBs, HBV DNA, PreS1 and other hepatitis B viral markers (HBV M) in the serum of patients infected with HBV.
METHODSHBV DNA was quantitatively detected using RT-PCR, LHBs, PreS1 and HBV M were analyzed by ELISA in totally 385 serum samples.
RESULTSThere was a significant difference between the positive rate of LHBs (86.97%) and PreS1 (49.5%) in the 307 serum positive for HBV DNA (P less than 0.05). There was a correlation between the levels of LHBs and the logarithm of HBV DNA (r=0.935). In the serum specimens of patients negative for HBeAg, there was no significant difference between the positive rate of LHBs (76.92%) and the HBV DNA (67.95%), but the positive rate of PreS1 (45.73%) was lower than that of LHBs or HBV DNA.
CONCLUSIONThere was a close correlation between the copies of HBV DNA and the levels of LHBs, both the positive rate and the coincidence rate of LBHs and HBV DNA were higher than those of PreS1. LHBs can reflect the replication status of HBV.
Adolescent ; Adult ; Aged ; DNA, Viral ; blood ; genetics ; Female ; Hepatitis B ; blood ; diagnosis ; Hepatitis B Surface Antigens ; blood ; Hepatitis B virus ; genetics ; immunology ; Humans ; Male ; Middle Aged ; Protein Precursors ; blood ; Reverse Transcriptase Polymerase Chain Reaction ; Sensitivity and Specificity ; Viral Matrix Proteins ; blood ; Young Adult
6.Comparison of BISAP, Ranson's, APACHE Ⅱ and CTSI scores in evaluating the severity of acute pancreatitis
Jinyan ZOU ; Jun LIN ; Sanfeng YI ; Qin XIANG ; Jian SHANG ; Bing XIA ; Changsheng DENG
Chinese Journal of Digestive Surgery 2014;13(1):39-43
Objective To investigate the value of the bedside index for severity in acute pancreatitis (BISAP),Ranson's,APACHE Ⅱ and computed tomography severity index (CTSI) scoring system in evaluating the severity of acute pancreatitis.Methods The clinical data of 385 patients with acute pancreatitis who were admitted to the Zhongnan Hospital of Wuhan University from 2005 to 2011 were retrospectively analyzed.The values of 4 scoring systems including BISAP,Ranson's,APACHE Ⅱ and CTSI in predicting the incidences of severe acute pancreatitis,local complications and death were investigated by Chi-square test and receiver operating characteristic curv e.Odds ratio (OR) was calculated.The differences of areas under the curves (AUC) were analyzed using the Z test.Results The incidences of severe acute pancreatitis,local complications and mortality of patients with BISAP score ≥ 3 were 64.4% (56/87),16.1% (14/87) and 8.0% (7/87),which were significantly higher than 13.4% (40/298),6.4% (19/298) and 0.3 % (1/298) of patients with BISAP score ≤ 2 (x2 =93.4,8.1,19.7,P < 0.05).The incidences of severe acute pancreatitis,local complications and mortality of patients with Ranson's score≥3 were 52.7% (48/91),22.0% (20/91) and 7.7% (7/91),which were significantly higher than 16.3% (48/294),4.4% (13/294) and 0.3% (1/294) of patients with Ranson's score ≤2 (x2 =49.2,27.3,18.5,P <0.05).The incidences of severe acute pancreatitis,local complications and mortality of patients with APACHE Ⅱ score ≥ 8 were 46.6% (27/58),20.7% (12/58) and 8.6% (5/58),which were significantly higher than 21.1% (69/327),6.4% (21/327) and 0.9% (3/327) of patients with APACHE Ⅱ score≤7 (x2 =17.0,12.8,14.4,P <0.05).The incidences of severe acute pancreatitis,local complications and mortality of patients with CTSI score ≥4 were 51.4% (19/37),51.4% (19/37),16.2% (6/37),which were significantly higher than 22.2% (77/347),4.0% (14/347),0.6% (2/347) of patients with CTSI score≤3 (x2 =15.1,95.3,40.1,P < 0.05).The sensitivity,specificity,positive and negative predictive values of BISAP were 58%,89%,64%,86%,respectively,and the AUC was 0.848,which were significantly higher than the other 3 systems (Z =2.02,4.22,4.78,P < 0.05).The sensitivity,specificity,positive and negative predictive values of CTSI were 58%,95%,51% and 96%,respectively,and the AUC was 0.926,which was significantly higher than the other 3 systems (Z =3.99,3.24,4.06,P < 0.05).The sensitivity,specificity,positive and negative predictive values of BISAP were 88%,79%,8% and 100%,respectively,and the AUC was 0.855,with no significant difference compared with the other 3 systems (Z =0.81,0.03,0.14,P > 0.05).Conclusions The accurate rate of BISAP in predicting the severe acute pancreatitis is higher than Ranson's,APACHE Ⅱ and CTSI.The accurate rate of CTSI in predicting the incidence of local complications is higher than the other 3 systems.There is no significant difference of the 4 systems in predicting the mortality.The BISAP scoring system is helpful in early diagnosis of severe acute pancreatitis,and making the individualized treatment plan,thus improving the prognosis of patients.
7.Establishing scientific evaluation view to promote traditional Chinese medicine
Xue-Min GAO ; Jian-Jun ZHANG ; Chun WANG ; Jing-Xia WANG ; Hai-Ying TONG ;
China Journal of Traditional Chinese Medicine and Pharmacy 2005;0(11):-
Traditional Chinese Medicine(TCM) makes great contributions to the prosperous growth and people's health.But understanding deviation and imperfect evaluation system of TCM affect the healthy development of TCM.Clinic practice is the motive power of TCM,and curative effect is the key of TCM researches,and the scientific evaluation system is the safeguard for a healthy development of TCM.So we should focus on clinical researches of stubborn diseases and emergency cases to satisfy social demand and upgrade the position of TCM in the medical system.At the same time,functional disease must be explored to show the advantage of TCM.Our mission is to establish a scientific objective evaluation system to accurately understand TCM and take it as the turning point to give an impetus to theoretical breakthrough of the basic studies to promote an overall and healthy development of TCM.
8.Expression of interleukin-13 receptor and its relationship to proliferation activity of human gliomas
Zhi-Bai XIA ; Xin-Jian WU ; Tie-Wei QI ; Hai-Jun WANG ; Zheng-Song HUANG ;
Chinese Journal of Microsurgery 2006;0(06):-
Objective To explore the gene expression of interleukin-13 receptor (IL-13R)?2 and its relationship to proliferation activity of human gliomas.Methods The gene expression of IL-13R?in 50 hu- man gliomas,2 malignant human glioma cell lines and 6 normal brain tissues were studied by RT-PCR.Ki-67 labeling index (Ki267 LI) of all sample were detecteded by immunohistochemical staining.Results Only one normal brain tissues expressed very low IL-13R?2 mRNA,whereas 35 (70%) of 50 human brain tumors expressed 1L-13R?2 mRNA.The positive rate and expression level of IL-13R?2 mRNA were increased with the ascending of WHO tumor grade.(former:rs=0.87;letter:rs=0.69,P<0.01).The difference of posi- tive rate and expression level of IL-13R 2?mRNA between the low grade and high grade tumors was statistical- ly significant,the proliferation activity of gliomas evaluated by Ki-67LI (Ki-67 Labeling Index,Ki-67LI) was positively correlated with IL-13R?2 gene expression and the tumor grade.Conclusion In human cerebral gliomas,IL-13R?2 genes may play an role in the malignant progression.The expression level of malignancy in molecular level and selecting the target of gene therapy.
9.A pharmacodynamic study of intervals of Mitomycin C treatment on human bladder cancer cell line in vitro
Yun-Xia WAN ; Jun TIAN ; Jian-Hui MA ; Han-Zhong LI ; Yan-Fen LI ; Wei SHI ;
Cancer Research and Clinic 2006;0(11):-
Objective To study the relationship between the intervals of Mitomycin C treatment and cytotoxicity, apoptosis and drug resistance for bladder cancer cells.Methods The bladder transitional cell cancer line BIU-87 was treated for two hours every time for five times with intervals of 24, 48, 72 and 96 hours respectively.Cytotoxicity was measured by MTT.p53,bcl-2,Bax and p170 expression were analyzed by Western blot.Results The IC_(50)(?g/ml)were 4.41,0.71,2.83,4.51and 6.16 with treatment intervals of 24, 48, 72 and 96 hours respectively, p53 and bcl-2 were significantly down-regulated and bcl-2/Bax was re- duced at 24 hour treatment interval but not changed at 48,72 and 96 hour intervals,p170 was not detected at 24 hour treatment interval but increasingly expressed at 48,72 and 96 hours intervals.Conclusion The in- terval of Mitomycin C treatment is closely related with cytotoxieity and apoptosis and drug resistance of blad- der cancer cells.The intervals of intravesical instillations may play an important role in the effect of chemotherapy.
10.Ligustrazine fought against cisplatin induced ototoxicity in guinea pigs.
Xian-Chang SUN ; Li-Xia SUN ; Xian-Jun SHI ; Song-Jian KANG
Chinese Journal of Integrated Traditional and Western Medicine 2014;34(8):982-986
OBJECTIVETo study the protective effect of Ligustrazine Injection (LI) against cisplatin-induced ototoxicity and to explore its mechanism.
METHODSThirty healthy adult guinea pigs were randomly divided into three groups, 10 in each group, i.e., the normal control group, the cisplatin group, and the LI group. Guinea pigs in the normal control group were intraperitoneally injected with normal saline at 3 mL/kg for 7 consecutive days. Those in the cisplatin group were intraperitoneally injected with cisplatin at 3 mg/kg for 7 consecutive days. Those in the LI group were intraperitoneally injected with LI at 140 mg/kg for 7 days, but cisplatin (3 mg/kg) was intraperitoneally injected from the opposite side starting from the 4th day. Brainstem auditory evoked potential (BAEP) was performed in all animals before and after injection. All animals were sacrificed after the final testing under anesthesia and their cochleas collected. Half the cochleas of each group were collected for silver nitrate staining of cochlear basilar membrane stretched. The other half the cochleas of each group made into paraffin sections to observe the apoptosis of cochlea cells by TUNEL method and the expression levels of c-Jun detected by immunohistochemistry.
RESULTSThere was no statistical difference in the difference of BAEP threshold among the 3 groups before injection (P > 0.05), but the BAEP threshold increased in the cisplatin group and the LI group (P < 0.05). Besides, it was higher in the cisplatin group (P < 0.05). In the cisplatin group, most hair cells were missing, spiral ganglion cells obviously decreased, more TUNEL positive cells occurred, and the expression of c-Jun was stronger. But the aforesaid impairment in the LI group was obviously lessened (P < 0.05).
CONCLUSIONSLI showed certain antagonist effect on cisplatin-induced ototoxicity. Its mechanism might be associated with scavenging oxygen radicals of the cochlea tissue, improving the microcirculation, and fighting against apoptosis.
Animals ; Apoptosis ; drug effects ; Cisplatin ; toxicity ; Cochlea ; drug effects ; metabolism ; pathology ; Evoked Potentials, Auditory, Brain Stem ; Guinea Pigs ; Pyrazines ; pharmacology ; Reactive Oxygen Species ; metabolism