Objective To construct antibody library in patient with pemphigus vulgaris using phage display antibody technique. MethodsTotal RNA was extracted from B cells of patients with pemphigus vulgaris(PV) using polymerase chain reaction,immunoglobulin VHand VL geneswere amplified by specific primer of HuVHBACK, HuJHFOR, HuVkBACK and HuJkFOR. The VH and VL genes were then cloned into a phage vector and expressed on the phage surface as a fusionprotein with product of gene Ⅲ.ResultsThe antibody phage library of single chain antibodies of patients with PV was constructed. ConclusionWe have obtained EC1-2 and EC3-4 purified protein of Dsg3 from constructed recombinant plasma in our laboratory which will facilitate the study on Dsg3 antibodies.

Paclitaxel is a kind of monomer diterpene compound extracted from the taxus chinensis, which has good anti-cancer activity. It is widely used in the treatment of breast cancer, ovarian cancer, lung cancer and other cancers. It also has been listed as the first-line chemotherapy medicine on breast cancer and ovarian cancer. However, the drug resistance is the main obstacle to clinical application similar to other kinds of chemotherapy medicine. The low toxicity and high efficient traditional medicine monomer, represented by curcumin, has become the research focus on reversing paclitaxel resistance. This article summarized the research on synergy between curcumin and paclitaxel, with a purpose to provide references for finding clinical assistant chemotherapeutic medicine.

Small RNA (miRNA) can regulate post-transcriptional level of mRNA. miRNA is closely related to the occurrence and development of a variety of human diseases such as gastric cancer, lung cancer, breast cancer, ovarian cancer, prostate cancer, pancreatic cancer, liver cancer, etc. Many kinds of miRNAs in human cancers are significantly abnomadly expressed. miRNA can be regarded as a marker for many human cancers diagnosis.

Objective To prepare a peptide monoclonal antibody (McAb)against human papillomavirus 18E6 separately,and identify its specificity and pathogenicity.Metheds The advantage epitope peptide was designed and synthesized by ABCpred and Bcepred,and then used to immunize BALB/c mice after coupling with bovine serum albumin (BSA).And the McAb was prepared by hybridoma technique.HPV18E6 gene was amplified from cervical swab specimen containing HPV18 and insert-ed into expression vector pET-28a.The constructed recombinant plasmid was transformed to E.coli BL21(DE3)for expres-sion under induction of isopropyl thio-β-D-galactoside.The expressed protein was used to identified the McAb had been pre-pared.Results The hybridoma cell lines could constantly produce MAbs against HPV18E6 peptides.Sequencing proved that recombinant plasmid pET-28a-HPV18E6 was constructed correctly.Western blotting showed that the anti-HPV18E6 pep-tides antibody could specifically recognize HPV18E6.Conclusion A monoclonal antibody against the advantage epitope pep-tide of human papillomavirus 18E6 prepared could specifically recognize HPV18E6 specifically.

Background and purpose: Ursolic acid is widely present in spica prunellae, hedyotis diffusa and other heat antidotes. The growth of a variety of tumor cells can be inhibited and induced apoptosis by ursolic acid.This study was aimed to investigate the effect and possible mechanisms of UA on inducing apoptosis of human gastric carcinoma BGC823 cells. Methods: The MTT assay was used to detect the antiproliferative effect of UA on BGC823 cells. Flow cytometry was used to detect cell cycle and apoptosis of BGC823 cells. The expression level of bcl-2 and bax gene was investigated by real time-polymerase chain reaction (real time-PCR). Results: UA inhibited the proliferation of BGC823 cells in a dose and time-dependent way. After treatment by UA for 24. 48 and 72 h, the IC_(50) of BGC823 was 36.88, 34.72, and 32.18 μmol/L, respectively. UA could signifcantly induce apoptosis of BGC823 cells and block cells at G_2/M phase. UA could increase the expression of bax gene and decrease the expression of bcl-2 gene in a dose and time-dependent way. Conclusion: UA could induce apoptosis and inhibit the proliferation of BGC823 cells in a dose and time-dependent way. It could arrest cell cycle of BGC823 cells at G_2/M phase. Its mechanisms might be associated with the up-regulation of bax gene and down-regulation of bcl-2 gene.

Objective: To clarify that protection of Tongxinluo against myocardial ischemia/reperfusion injury relates to enhancing endothelial barrier in diabetic rats.
Methods: A total of 32 Zucker diabetic rats were randomized into 4 groups:Sham group, Model group, Insulin group and Tongxinluo group, n=8 in each group. In addition, there was a Control group containing 8 non-diabetic Zucker rats. Myocardial ischemia/reperfusion injury model was established by a 45-min ischemia and 3-h reperfusion protocol. The size of infarction was detected by pathological staining, the microvascular permeability was examined by Miles assay to obtain the lfuorescein isothiocyanate concentration in myocardial tissue, the Triton X-100 soluble and insoluble VE-cadherin was measured by membrane protein extraction and western blot analysis.
Results:The size of infarction in Model group was obviously higher than that in Control group (55.2 ± 1.4)%vs (36.2± 1.3)%,P<0.05 and the lfuorescein isothiocyanate concentration was also higher in Model group than Control group. Both insulin group and Tongxinluo group had the similar size of infarction with Model group (36.8 ± 1.2)%vs (38.7 ± 1.1)%, P>0.05. Both insulin group and Tongxinluo group got lower lfuorescein isothiocyanate concentration (all P<0.01) and VE-cadherin internalization (all P<0.05) than that in Model group.
Conclusion: Protection of Tongxinluo against myocardial ischemia/reperfusion injury in diabetic rats is as effective as
insulin, but the effect is independent of reducing blood glucose and may be related to enhancing endothelial barrier.

Myeloid differentiation factor 88 (MyD88) plays an important role in tumorigenesis,development and malignant transformation,also participates in the microenvironment,proliferation,apoptosis,invasion,metastasis and tumor resistance.MyD88 may serve as a new and meaningful therapeutic target,which can promote the growth and development of tumors by regulating multiple signaling pathways and enhance the drug resistance of tumor cells.

OBJECTIVETo explore the genetic etiology of a pedigree affected with hereditary retinitis pigmentosa.

METHODSHigh-throughput DNA sequencing was used to analyze the sequences of 173 genes associated with hereditary eye diseases in the proband. Suspected mutation was verified with PCR amplification and Sanger sequencing.

RESULTSThe proband was found to have carried a c.570_571 ins GAAGATGCTGT insertional mutation in the RP2 gene located on the X chromosome. All female carriers of the pedigree were heterozygous, while all affected males were hemizygous for the same mutation.

CONCLUSIONThe inheritance pattern of this retinitis pigmentosa pedigree was X-linked recessive. The c.570_571 ins GAAGATGCTGT insertional mutation of the RP2 gene probably underlies the disease.

Eye Proteins ; genetics ; Female ; Genetic Diseases, X-Linked ; genetics ; High-Throughput Nucleotide Sequencing ; Humans ; Intracellular Signaling Peptides and Proteins ; genetics ; Male ; Membrane Proteins ; genetics ; Pedigree ; Pregnancy ; Prenatal Diagnosis ; Retinitis Pigmentosa ; genetics

OBJECTIVE: To explore the genetic basis for a pedigree affected with congenital sensorineural deafness. METHODS: High-throughput sequencing was carried out to analyze the coding regions of 415 genes associated with hereditary deafness in the proband. Suspected variants were verified by PCR amplification and Sanger sequencing of her parents and sister. RESULTS: The proband was found to have carried a heterozygous c.5131G>A (p.Val1711Ile) variant of the CDH23 gene and a heterozygous c.2884C>T(p.Arg962Cys) variant of the PCDH15 gene, which were respectively inherited from her mother and father. Her sister (with normal hearing) was also heterozygous for the c.5131G>A (p.Val1711Ile) variant of the CDH23 gene but not the c.2884C>T (p.Arg962Cys) variant of the PCDH15 gene. Based on the guidelines of the American College of Medical Genetics and Genomics, both variants were predicted to be likely pathogenic (PS1+PM2+PP3+PP4). CONCLUSION The c.5131G>A (p.Val1711Ile) variant of the CDH23 gene and c.2884C>T (p.Arg962Cys) variant of the PCDH15 gene probably underlay the pathogenesis of Usher syndrome type 1D/F in this pedigree.
Female ; Heterozygote ; High-Throughput Nucleotide Sequencing ; Humans ; Mutation ; Pedigree ; Usher Syndromes/genetics*

OBJECTIVE: To explore the genetic basis of a Chinese pedigree affected with progressive non-syndromic sensorineural hearing loss. METHODS: High-throughput DNA sequencing was carried out to analyze 415 genes associated with hereditary deafness in the proband. Sanger sequencing was carried out to verify the suspected variants among her family members. RESULTS: The proband was found to carry a heterozygous c.842T>A (p.Ile281Asn) variant of the POU4F3 gene. The same variant was found among all other patients from the pedigree including the proband's mother, brother, aunt and maternal grandfather, but not among those with normal hearing. Based on the standards and guidelines of the American College of Medical Genetics and Genomics and the Association for Molecular Pathology, the c.842T>A(p.Ile281Asn) variant of the POU4F3 gene was predicted as likely pathogenic (PM2+PM5+PP1+PP3+PP4). CONCLUSION A Chinese pedigree affected by a rare type autosomal dominant deafness-15 (DFNA15) due to a novel c.842T>A (p.Ile281Asn) variant of the POU4F3 gene was identified. The result has facilitated genetic counseling and risk assessment for the pedigree.
China ; Deafness/genetics* ; Female ; Genetic Testing ; Hearing Loss, Sensorineural/genetics* ; Humans ; Male ; Mutation ; Pedigree