OBJECTIVETo explore efficacy enhancing and detoxification roles of Jiedu Quyu Zishen Recipe (JQZR) in treating systemic lupus erythematosus (SLE) by studying its effect on Toll like receptor 9 (TLR9) signal pathway of murine macrophage cells after JQZR stimulated CpG oligodeoxynucletide (CpG ODN).

METHODSMurine macrophage cells in vitro cultured were randomly divided into 4 groups, i.e., the blank serum group, the CpG ODN stimulus group, the CpG ODN + dexamethasone group, the CpG ODN + medicated serum group. Murine macrophage cells were collected after 24-h intervention. The expression of TLR9, myeloid differentiation factor 88 (MyD88), NF-KB, IFN-α mRNA were analyzed by RT-PCR. The expression of TLR9 and NF-κB protein were analyzed by Western blot. Changes of the NF-KB transcriptional activity were assayed by Dual-Luciferase reporter assay system.

RESULTSmRNA expressions of TLR9, MyD88, NF-κB, and IFN-α, protein expressions of TLR9 and NF-κB, and NF-κB transcriptional activities were enhanced, showing statistical difference when compared with those of the blank serum group (P <0. 05, P <0. 01). Compared with the CpG ODN stimulus group, mRNA expressions of MyD88, NF-κB, and IFN-α, the protein expression of NF-κB and the NF-κB transcriptional activities decreased in the CpG ODN + dexamethasone group with statistical difference (P <0. 01). Compared with the CpG ODN stimulus group, mRNA expressions of TLR9, MyD88, NF-κB, and IFN-α, protein expressions of TLR9 and NF-κB, and NF-κB transcriptional activities were decreased in CpG ODN+ medicated serum group with statistical difference (P <0. 01).

CONCLUSIONEfficacy enhancing and detoxification roles of JQZR in treatment of SLE might be realized through regulating TLR9 signal pathways.

Animals ; Cell Line ; Drugs, Chinese Herbal ; pharmacology ; Humans ; Macrophages ; metabolism ; Mice ; Myeloid Differentiation Factor 88 ; NF-kappa B ; RNA, Messenger ; Signal Transduction ; Toll-Like Receptor 9 ; metabolism

AIM: To examine the effect of traditional chinese medicine recipe, Taoren Honghua(semen persicae-flos carthami) decoction, on hyperlipidemia without symptom. METHODS: The plasma TC, TG, LDL, HDL, apolipoprotein(Apo) A, Apo B of the patients with hyperlipidemia without symptom were measured using automatic analyzer (shimadiu CL-7200), the production of nitric oxide(NO) was detected by Greiss reaction, and SOD activity and MDA formation were examined using o-trihydroxy benzene and barbituric methotheds, respectively. RESULTS: After oral administration of Taoren Honghua decoction, the plasma levels of TC, TG, LDL, and MDA of the patients were markedly decreased, however, the plasma levels of ApoA, HDL, SOD and NO were significantly increased and almost no change was detected in the plasma level of Apo B. In control group, it was found that although the plasma level of TC, TG and LDL were decreased ( P

Objective To clone and express the encoding sequence of glyceraldehydes-3-phosphate dehydrogenase(GAPDH)from periodic Brugia molayi(Bm).Methods Total RNA was extraeted from periodic Brugic malayi.The BmGAPDH gene was amplified by RT-PCR.The PCR product was cloned and then subeloned into pcDNA3.1(+)vector.The recombinant plasmids were screened and identified by digestion with restriction enzyme and PCR amplification,and were transformed into COS-7 cell subsequently.The expressed protein was identified by SDS-PAGE.Results BmGAPDH mRNA was highiy expressed in transfected COS-7 cell.The deduced amino acid sequence was identical with that of BmGAPDH.The recombinant pnotein wag about Nr 43 000.Conclusion The recombinant plasmid peDNA3.1(+)-BmGAPDH has been constructed and the protein has been expressed correctly.

Objective To discuss the clinical features of cerebral schistosomiasis. Methods The clinical data of fourteen patients with cerebral schistosomiasis from March 2010 to March 2016 were collected and analyzed retrospectively. Results The schistosomiasis immunological tests of sera and cerebrospinal fluids from the fourteen patients were all positive. Eosinophils increased in ten cases,and the proportion was 5.1％-60.3％. Schistosoma eggs were found in seven cases by the fecal Kato-Katz method. Fourteen cases were all infected with Schistosome japonicum. Twelve cases were diagnosed as chronic type,and two cas-es as acute type. Thirteen patients received medical treatment,of which twelve were cured,and one improved. One patient re-ceived the surgical resection of the lesion. Conclusion The clinical manifestations of cerebral schistosomiasis mainly include seizure,headache,dizziness and fever. In the enhanced head magnetic resonance imaging(MRI),the lesions are clustered and merged into lumps,which is the characteristic image of cerebral schistosomiasis japonica. The praziquantel treatment can achieve a good prognosis.

Objective To clone and sequence the cysteine protease inhibitor gene of periodic Brugia malayi(BmCPI) and predict B-cell epitopes in amino acide sequence of BmCPI in order to provide basis for further study the expression of BmCPI and its function. Methods Total RNA was extracted from periodic Brugia malayi.A couple of specific primers were designed on the basis of known sequences of cysteine protease inhibitor gene from BmCPI. The desired gene was amplified by PCR technique from cDNA. The PCR products were purified and cloned into plasmid pGEM-T by T-A cloning method, transformed into Escherichia coli(E, coli) strain DH5α. The recombinant plasmids were screened and identified by digestion with restriction enzyme and PCR amplification. Five parameters and methods were used to predict B-cell epitopes in amino acide sequence of BmCPI. Results For RT-PCR, a specific band of around 621 bp was amplified. The same band was obtained by double restriction of recombinant plasmids or PCR using recombinant plasmid as template. The result of DNA sequencing showed that BmCPI shares 99% nucleotide sequence identity with that of published sequence. It showed that B-cell epitopes were probably at or adjacent to 23 - 32, 50 - 79 and 117 - 126 in its amino acide sequence. Conclusions pGEM-BmCPI is successfully constructed and sequenced, anticipated objective is reached and conditions is provided for further study of BmCPI expression and its function.

OBJECTIVETo evaluate the treatment of recurrent acute cholangitis with hepatolithiasis.

METHODSWe retrospectively analyzed the clinical data of patients with recurrent acute cholangitis who were treated in Peking Union Medical College Hospitals emergency department from January 1998 to December 2008.

RESULTSTotally 408 patients underwent surgery, of which 167 patients received emergency operations and 241 underwent selective operations after medication and interventional treatment. The incidence of complications was 6.4% among those who received emergency operations and 3.2% among selective operations. The 30-day mortality rate of selective operations was zero.

CONCLUSIONAlong with the progress of percutaneous cholangiographic drainage and endoscopic retrograde cholangiopancreatography, selective operations have been increasingly applied for acute cholangitis with notably low complications and postoperative death.

Adolescent ; Adult ; Aged ; Aged, 80 and over ; Cholangitis ; etiology ; surgery ; Cholelithiasis ; complications ; surgery ; Female ; Humans ; Male ; Middle Aged ; Retrospective Studies ; Treatment Outcome ; Young Adult

OBJECTIVETo study the prognostic identification of lymph node negative breast carcinoma by quantitative pathologic technique.

METHODSSeveral morphometrical parameters, DNA content of cell nuclei were detected by means of a quantitative pathologic technique on 102 patients with lymph node negative invasive breast duct carcinoma. The effects of potential prognostic factors of lymph node negative breast cancer patients were assessed by Cox's proportional hazards regression model.

RESULTSDNA index, minimal diameter of nuclei, area of nuclei, maximal diameter of nuclei, and the perimeter of nuclei are important factors to influence the prognosis.

CONCLUSIONQuantitative pathologic technique combined with valid statistical methods, as an objective means of assessing prognosis, may reliably improve the outcome in lymph node negative breast cancer.

Adult ; Aged ; Breast Neoplasms ; pathology ; DNA, Neoplasm ; analysis ; Female ; Humans ; Lymphatic Metastasis ; Middle Aged ; Multivariate Analysis ; Prognosis

OBJECTIVE: To explore the effect and toxicity profile of recombinant human interleukin 11(rhIL-11) on the platelet after hematopoietic stem cell transplantation in patients with leukemia. METHODS: Twenty-four patients with acute or chronic leukemia treated by allogeneic peripheral blood stem cell transplantation (PBSCT) were randomly divided into a test group and a control group. The patients in the test group were treated with rhIL-11 since the 13th day after PBSCT (1.5mg/d),while the control group were given symptomatic treatment. RESULTS: The average time for the platelet to recover to the level of 20 x 10(9)/L was 20.8 days in the test group, and 26.0 days in control group respectively, there was significant difference (P<0.01). The average time for the platelet to recover to the level of 50 x 10(9)/L was 25.7 days in the test group, and 32.3 days in the control group respectively, there was also significant difference (P<0.01). The average time for the platelet transfusion was 2.2 in the test group, 4.1 in the control group, and there was significantly different (P<0.01). The average number of megakaryocytes was 12.2 in the test group, 4.8 in the control group on 30th day after the transplantation,and there was significant difference(P<0.01). The main side effects of rhIL-11 were nausea, vomit, debility, headache, dizzy and pain of injection site, and the degree was all Iapproximately II grade. CONCLUSION rhIL-11 has definite recuperative effect on the recovery of the platelet after PBSCT. There is little side effect, and it can be accepted.
Adolescent ; Adult ; Female ; Hematopoietic Stem Cell Transplantation ; Humans ; Interleukin-11 ; genetics ; therapeutic use ; Leukemia ; blood ; surgery ; Leukemia, Myelogenous, Chronic, BCR-ABL Positive ; blood ; surgery ; Leukemia, Myeloid, Acute ; blood ; surgery ; Male ; Middle Aged ; Platelet Count ; Recombinant Proteins ; therapeutic use ; Thrombocytopenia ; drug therapy ; Treatment Outcome

OBJECTIVE: To construct two types of Wnt-inducible secreted protein 3(WISP3) gene's mutants(1000T/C,840delT) found in spondyloepiphyseal dysplasia tarda with progressive anthopathy (SEDT-PA) patients, and to observe their expression in COS-7 cells. METHODS: Full-length cDNA of wild type WISP3 gene(WT-WISP3) was amplified from human chondrocytes by RT-PCR, and site-directed mutagenesis was used to obtain full-length cDNAs of the mutated WISP3 genes(MUT1000T/C and MUT840delT). The recombined plasmids WT-WISP3/pcDNA3.1(+), MUT1000T/C/pcDNA3.1(+) and MUT840delT/pcDNA3.1(+) were transfected transiently into COS-7 cells by liposome-mediated method, and pcDNA3.1(+) vector was used as a control. The total RNA and protein of the transfected COS-7 cells were extracted after 48 hours of transfection. The expression of WISP3 gene in the transfected COS-7 cells was detected by semi-quantitative RT-PCR and Western blot. RESULTS: By restriction endonuclease analysis and sequencing, the sequence of MUT1000T/C and MUT840delT were consistent with that mutated in SEDT-PA, and the open reading frames matched with the vector sequence. Semi-quantitative RT-PCR and Western blot showed that the recombined plasmids were highly expressed in COS-7 cells. CONCLUSION WISP3 gene's mutants of SEDT-PA are successfully constructed by genetic recombination, and expressed in COS-7 cells, which lays the foundation for the further study on its molecular functions in SEDT-PA.
Animals ; Base Sequence ; CCN Intercellular Signaling Proteins ; COS Cells ; metabolism ; Chlorocebus aethiops ; Humans ; Insulin-Like Growth Factor Binding Proteins ; biosynthesis ; genetics ; Molecular Sequence Data ; Mutagenesis, Site-Directed ; Mutation ; Osteochondrodysplasias ; genetics ; metabolism ; Transfection

OBJECTIVETo explore the necessity of large-scale screening of mitochondria DNA (mtDNA) A1555G mutation for prevention of aminoglycoside antibiotic induced deafness in newborns.

METHODSOne thousand blood filter samples were collected from neonates born in July 2008 in Shenzhen. DNA was extracted with Chelex-100 Resin and amplified by PCR. The mtDNA A1555G mutation was determined by denaturing high-performance liquid chromatography(DHPLC) for PCR products. The positive frequency was calculated.

RESULTSThe mitochondrial DNA A1555G mutation was detected in 2 cases of 1000 neonates. The frequency of mutation was 0.2%.

CONCLUSIONThere is a high frequency of mtDNA A1555G mutation in neonates, the large-scale screening of mtDNAA1555G mutation in newborns might detect the individuals sensitive to aminoglycoside antibiotic, which is helpful to guide a rational medication for newborns and the maternal relatives at high-risk. Furthermore, it might be useful to prevent aminoglycoside antibiotic induced deafness.

Base Sequence ; DNA Mutational Analysis ; methods ; DNA, Mitochondrial ; genetics ; Female ; Humans ; Infant, Newborn ; Male ; Polymerase Chain Reaction