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.

Acute Disease ; Adult ; Cholecystitis, Acute ; diagnosis ; pathology ; Diagnostic Errors ; Female ; Humans ; Myocarditis ; diagnosis ; pathology

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.

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

OBJECTIVETo study the effects of arsenic trioxide (As2O3) on the proliferation and the migration force of human breast cancer SKBR-3 cell and the expression of Notch1.

METHODSSKBR-3 cells were cultured with different concentrations of As2O3 for 24 h and with the final concentration of 8 micromol/L for 24, 48, and 72 h. The effects of As2O3 on the cell proliferation of SKBR-3 were detected by MTT assay. The effects of the migration force of SKBR-3 cells were detected by Transwell. The expression of Notch1 mRNA was detected using reverse transcription polymerase chain reaction (RT-PCR). The expression of Notch1 protein was detected using Western blot.

RESULTSAs2O3 could significantly inhibit the proliferation of SKBR-3 cells in a concentration- and time-dependent manner (P < 0.05). It also could inhibit the migration force of SKBR-3 cells (P < 0.05). Results of RT-PCR and Western blot showed that Notch1 mRNA and protein levels obviously decreased (P < 0.05).

CONCLUSIONAs2O3 could inhibit the expression of Notch1 and the cell proliferation and the migration force of SKBR-3 cells, which primarily revealed that As2O3 might affect the biological behavior of human breast cancer cells possibly through Notch1 signaling pathway, thus providing theoretical and experimental bases for treating breast cancer by arsenic.

Arsenicals ; pharmacology ; Breast Neoplasms ; metabolism ; Cell Line, Tumor ; Cell Movement ; drug effects ; Cell Proliferation ; drug effects ; Female ; Humans ; Oxides ; pharmacology ; RNA, Messenger ; genetics ; Receptor, Notch1 ; metabolism ; Signal Transduction ; drug effects

Humans ; Stents ; Tympanic Membrane ; Tympanic Membrane Perforation ; surgery

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

Objective To analyze the diagnosis and prognosis value of abdominal CT scans in patients with acute pancreatitis complications.Methods 151 cases with acute pancreatitis were selected.The relationship between abdominal CT performances and the common complications and death in patients was analyzed.The prognosis of patients with different Balthazar CT severity index (CTSI) grade was compared.Results The complication rate of patients with CT signs of fatty liver, pleural effusion, liver gap effusion, adrenal gland involvement (AGI), penirenal space involvement (PSI) and gastric bare area involvement (GBAI) was significantly higher than that of patients with negative CT findings above (P<0.05).With CTSI grading increasing, the patient''s fasting time, heating time, hospital stay, recovery time of blood amylase were extended, and the incidence of pseudo cyst, transit surgery, organ failure and death was gradually increasing (P<0.05).Conclusion Severe fatty liver, AGI, GBAI, PSI and liver gap effusion are risk factors for acute complications and death in patients with pancreatitis.

ObjectiveTo identify serum specific protein biomarkers for the early diagnosis of lymphoma patients by using surface enhanced laser desorption/ionization time of flight mass spectrometry (SELDI-TOF-MS)protein chip array.MethodsSELDI-TOF-MS technique and weak cation exchanger (WCX2)protein chip were used to detect the serum proteomic patterns of 96 patients with lymphoma (86 patients with non-Hodgkin lymphoma and 10 patients with Hodgkin lymphoma)and 30 normal control.The serum level of LDH were measured by biochemical methods,the serum level of β2-MG and CA125 were detected by enzyme-linked immunosorbent assay (ELISA).ResultsFour protein peaks pattern mass/charge ratio (M/Z) 6880,8564,8692 and 13751 were decreased in lymphoma patients,and they could be used to distinguish lymphoma from healthy people and other malignant tumors.Their sensitivity and specificity were 82.29 ％ and 80.00 ％,78.13 ％ and 73.30 ％,75.00 ％ and 80.00 ％,71.88 ％ and 83.30 ％ respectively in lymphoma patients.When it was used to early diagnosing lymphoma in stage Ⅰ or Ⅱ,the sensitivity of four protein peaks pattern (M/Z 6880,8564,8692 and 13 751) were 77.55 ％,71.43 ％,71.43 ％ and 67.35 ％respectively and the specificity of all four protein peaks pattern were 100.00 ％.Four protein peaks pattern sensitivity were higher than LDH,CA125 and β2-MG respectively.ConclusionApplication of SELDI-TOF-MS can be of great potential for the early diagnosis of lymphoma patients.

Objective To investigate the effects of family allergic history and dermatophagoides farina on the expressions of Th1/Th2 cytokine of cord blood and allergic disorders in infancy.Methods Ten mil cord blood were obtained from 34 neonates which 17 cases had family allergic history and 17 cases didn′t have.Cord blood mononuclear cells(CBMC) were isolated by gradient centrifugation with Ficoll and were cultured with phytohemagglutinin(PHA) or dermatophagoides farina for 48 hours in vitro.The expressions of interleukin(IL-4) and interferon(IFN-?) of the culture supernatant were detected by enzyme-linked immunosorbent assay(ELISA).Two groups were visited with telephone or clinical service every 1 or 2 months in 1 year follow-up survey.Results In no stimulation,the expressions of IL-4 of family allergic history and no family allergic history were(11.35?1.80) ng/L and(11.0?1.50) ng/L,respectively,there was no significant difference.The expressions of IFN-? were(9.55?1.47) ng/L and(10.19?1.37) ng/L,respectively,there was no significant difference also.In PHA stimulation,the expressions of IL-4 were(43.45?4.57) ng/L and(37.58?3.41) ng/L,respectively,there was significant difference.The expressions of IFN-? were(72.61?25.40) ng/L and(65.84?29.96) ng/L,respectively,there was no significant diffe-rence.In low density dermatophagoides farina stimulation,the expressions of IL-4 were(40.54?3.64) ng/L and(37.17?2.60) ng/L,respectively,which had significant difference.The expressions of IFN-? were(35.30?2.73) ng/L and(40.55?1.85) ng/L,respectively,which had significant difference.In high density dermatophagoides farina stimulation,the expressions of IL-4 were(43.50?3.19) ng/L and(39.55?4.13) ng/L,respectively,which had significant difference.The expressions of IFN-? were(39.40 ?5.21) ng/L and(40.94?2.96) ng/L respectively,which had no significant difference.Allergic diseases were happened in 7 cases of 13 cases with family allergic history and in 2 cases of 15 cases without family allergic history in 1 year follow-up except lost follow-up cases.There were significant difference in 2 groups.Conclusions Th2 cells of cord blood are relative dominant in neonates having family allergic history.Th2 cells relative dominant are more obvious in dermatophagoides farina stimulation.The neonates having family allergic history have a tendency to get allergic diseases in childhood.