OBJECTIVETo identify the expression characteristics of the 1700001022RIK (RIKEN cDNA 1700001022) gene in mice and explore its function by bioinformatic analysis.

METHODSUsing the expression profile of gene microarray, we detected the expression of a new testis-specific gene, 1700001022RIK, in mice. We analyzed its expression characteristics in the testis tissue and their changes in different developmental stages of the testis by RT-PCR, real-time RT-PCR, Western blot, and immunohistochemistry. We performed bioinformatic analysis using a bioinformatic software.

RESULTSThe 1700001022RIK gene was specifically expressed in the mouse testis in an age-dependent manner, most highly in the adult mice. The 1700001022RIK protein was mainly expressed in the spermatogonia, spermatocytes, and round spermatids of the adult mice. Bioinformatic analysis showed that the 1700001022RIK protein amino acid sequence had a high similarity in human and mice, which indicated that this gene was highly conserved in mammals.

CONCLUSION1700001022RIK is a testis-specific gene mainly expressed in the spermatogonia, spermatocytes, and round spermatids of seminiferous tubules, which might be involved in the regulation of spermatogenesis.

Age Factors ; Animals ; Blotting, Western ; Computational Biology ; DNA, Complementary ; Gene Expression ; Genomics ; Male ; Mice ; Molecular Chaperones ; genetics ; Seminiferous Tubules ; Spermatids ; Spermatocytes ; Spermatogenesis ; genetics ; Spermatogonia ; Testis

OBJECTIVETo observe the therapeutic effect of electroacupuncture for treatment of sudden hearing loss and to compare with western medicine therapy.

METHODSSixty cases were randomly divided into an electroacupuncture group and a medication group, 30 cases in each group. The electroacupuncture group was treated with electroacupuncture at Tinghui (GB 2), Yifeng (TE 17), Hegu (LI 4), Xiaxi (GB 43), Zhongzhu (TE 3), etc. and the medication group with intravenous dripping of 6% low molecule dextran 500 mL with ATP and coenzyme A, and oral administration of Nimodipine, Gold Theragan. Whole blood specific viscosity, plasma specific viscosity, hematocrit and fibrinogen before and after treatment and their therapeutic effects were observed.

RESULTSThe total effective rate was 86.7 in the electroacupuncture group and 60.0% in the medication group with a significant difference between the two groups (P<0.05), the former being better than the latter; there were significant differences in whole blood specific viscosity, plasma specific viscosity, hematocrit and fibrinogen before and after treatment in the electroacupuncture group (P<0.05), and with no significant difference in the medication group (P>0.05) before and after treatment, and with a significant difference in whole blood specific viscosity, plasma specific viscosity and fibrinogen between the two groups (P<0.05).

CONCLUSIONElectroacupuncture has a significant therapeutic effect on sudden hearing loss, which is better than that of the medication, and the mechanism is possibly related with regulative action on indexes of blood rheology.

Acupuncture Points ; Adult ; Electroacupuncture ; Female ; Hearing Loss, Sudden ; therapy ; Humans ; Male ; Middle Aged

OBJECTIVETo generate a candidate HPV16 vaccine for experimental and therapeutical use for cervical cancer.

METHODSThe mutants of HPV16 early E6 and E7 genes were inserted into a vaccinia virus expression vector. A strain of recombinant vaccinia virus was constructed through homologous recombination.

RESULTSShowed that the mutant E6 and E7 genes were located at TK gene region of vaccinia virus Tiantan strain in a head to head orientation under the control of early/late promoters, H6 and 7.5K respectively. Studies in mice indicated that VmE6E7 could elicit specific antibodies against E6 and E7, and retarded or prevented tumor development in a proportion of C57 BL/6 mice challenged by syngeneic HPV16E6 and E7 transformed tumor cells.

CONCLUSIONSThe success in constructing VmE6E7 provides a basis for the further development of HPV16 therapeutic vaccine.

Animals ; Female ; Genes, Viral ; genetics ; Genetic Vectors ; genetics ; immunology ; Mice ; Mice, Inbred C57BL ; Mutation ; Neoplasms, Experimental ; prevention & control ; Oncogene Proteins, Viral ; genetics ; Papillomaviridae ; genetics ; Papillomavirus E7 Proteins ; Recombination, Genetic ; Repressor Proteins ; Transfection ; Vaccinia virus ; genetics ; immunology

OBJECTIVETo generate an HPV16 prophylactic vaccine candidate for cervical cancer.

METHODSHPV16 major capsid protein L1 gene and minor capsid protein L2 gene were amplified using PCR. These genes were mutated by PCR site-directed mutagenesis for removal of sequence motifs (TTTTTNT) which would cause transcription termination when expressed from a vaccinia virus early promoter, then inserted into a vaccinia virus expression vector. A strain replication-deficient recombinant vaccinia virus containing the mutant sequences was obtained through a homologous recombination and identified.

RESULTSThe nucleotide sequence remained the correct amino acid sequence of the L1 and L2 proteins after mutated. Full-length L1 and L2 proteins were generated in cells infected with the recombinant virus. The virus strain propagated at very low titer or could not reproduce in some kinds of cell derived from different human tissues.

CONCLUSIONSThe authors have generated a strain replication-deficient recombinant vaccinia virus expressing HPV16 L1 plus L2 proteins as an HPV16 prophylactic vaccine candidate for cervical cancer.

Capsid ; Capsid Proteins ; genetics ; Cell Line ; Cloning, Molecular ; Female ; Gene Expression ; Genetic Vectors ; Humans ; Oncogene Proteins, Viral ; genetics ; Papillomaviridae ; genetics ; physiology ; Papillomavirus Infections ; prevention & control ; Transfection ; Tumor Virus Infections ; prevention & control ; Uterine Cervical Neoplasms ; virology ; Vaccinia virus ; genetics ; physiology ; Virus Replication

OBJECTIVETo investigate the effect and mechanism (a selective cyclooxygenase-2 inhibitor) on invasive ability of human nasopharyngeal carcinoma (NPC) line CNE-2Z.

METHODSThe proliferation of NPC cells was examined by MTT assay. The invasive and migrating ability of NPC cells was detected with transwell chamber. E-cadherin protein expression was detected by immunocytochemistry and the expressions of Cox-2 and E-cadherin mRNA were analyzed by reverse transcription-polymerase chain reaction (RT-PCR).

RESULTSMTT showed that celecoxib inhibited CNE-2Z proliferation in dose-dependent manner, the survival rate of cells treated with 25, 50, 100 µmol/L celecoxib (x(-) ± s) for 24 h was (94.75 ± 1.34)%, (91.77 ± 2.70)%, (64.54 ± 1.20)%, respectively, and the survival rate of cells treated for 48 h was (88.41 ± 1.28)%, (78.84 ± 1.56)%, (52.46 ± 2.25)%, respectively, the concentration of 50% inhibition concentration of a substance (IC50) was 100 µmol/L, the difference was statistically significant between different concentration groups in the same time-point (respectively, F were 462.204 and 1328.306, P < 0.01). Treated with different concentrations of celecoxib (0, 25, 50 µmol/L) for 24, the cell numbers (x(-) ± s) through PVPF by tumor invasion assay were (263.7 ± 13.5), (185.3 ± 8.7) and (144.0 ± 8.2), the difference was statistically significant between the experimental and control group (F = 102.089, P < 0.01). Immunocytochemistry showed that celecoxib significantly induced the increase of E-cadherin protein expression, also with a dose-dependence in 0 µmol/L, 25 µmol/L, 50 µmol/L group was (21.7 ± 2.6), (28.7 ± 2.4), (40.3 ± 1.3), and 50 µmol/L group increased significantly (F = 78.637, P < 0.01). RT-PCR showed that celecoxib reduced the expression of Cox-2 mRNA expression in 25, 50 µmol/L group decreased significantly compared with the control group (respectively, t were 23.950 and 36.651, P < 0.01), but it enhanced the expression of E-cadherin mRNA expression in 25, 50 µmol/L group was significantly higher (respectively, t were 35.829 and 81.497, P < 0.01).

CONCLUSIONCelecoxib can inhibits the invasive ability of NPC cell line CNE-2Z, which possibly relates with the upregulated expression of E-cadherin.

Apoptosis ; drug effects ; Cadherins ; genetics ; Carcinoma, Squamous Cell ; metabolism ; pathology ; Celecoxib ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Gene Expression Regulation, Neoplastic ; Humans ; Nasopharyngeal Neoplasms ; metabolism ; pathology ; Neoplasm Metastasis ; Pyrazoles ; pharmacology ; Sulfonamides ; pharmacology

BACKGROUNDTo select the mutants of HPV type 16 E6 and E7 genes suitable for construction of vaccine for treatment of cervical cancer.

METHODSE6 and E7 genes were modified by site-directed mutagenesis. Several recombinant vaccina viruses were constructed by inserting the E6 or E7 mutants into the genome of vaccina virus Tiantan strain and employed to study their antigenicity.

RESULTSWestern blot assay showed that the E6 ?mutant? with substitution of Gly for Leu at amino acid site 50 and E7 mutant with substitution of Gly for Cys-24 and Glu-26 had no effect on their stability and antigenicity, but change of the Cys at position 91 of E7 dramatically reduced its stability and antigencity. Conclusion The results confirmed that the Zinc-finger structure at the E7 C-terminal? plays an important role in the integrity and stability of E7 protein.

Animals ; Antibodies, Viral ; biosynthesis ; Female ; Mice ; Mice, Inbred C57BL ; Mutagenesis, Insertional ; Oncogene Proteins, Viral ; genetics ; immunology ; Papillomaviridae ; genetics ; Papillomavirus E7 Proteins ; Repressor Proteins ; Vaccinia virus ; immunology ; Zinc Fingers

Objective To evaluate the effects of interleukin-22 (IL-22) on the expression of CC chemokine ligand 27 (CCL27) in human epidermal keratinocytes,and to explore its mechanism.Methods Immunohistochemical study was performed to determine the expression of CCL27 in skin lesions of 10 patients with psoriasis and skin tissues of 5 healthy controls.Cultured HaCaT cells were divided into 8 groups:control group treated with PBS,5 IL-22 groups treated with 12.5,25,50,100 and 200 μg/L IL-22 respectively,2 signaling pathway inhibition groups treated with 50 μrmol/L AG490 (JAK2/STAT3 signaling pathway inhibitor) or PD98059 (MAPK-ERK1/2 signaling pathway inhibitor) for 2 hours followed by the treatment with 50 μg/L IL-22 in the 5％ CO2 incubator at 37 ℃.After 24-hour cultivation,total proteins were extracted,and culture supernatants were collected,and both Western blot analysis and enzyme-linked immunosorbent assay (ELISA) were performed to determine the expression of CCL27.Results Immunohistochemical study showed that the expression of CCL27 was significantly higher in the skin lesions of the patients with psoriasis than in the skin tissues of the healthy controls.Western blot analysis revealed that the protein expression of CCL27 in the 12.5-,25-,50-,100-and 200-μg/L IL-22 groups was 0.491 ± 0.013,0.620 ± 0.019,0.623 ± 0.014,0.802 ± 0.052 and 1.138 ± 0.013 respectively,which were all higher than that in the control group (0.413 ± 0.013,all P ＜ 0.01).The expression of CCL27 was significantly lower in the IL-22 + AG490 group (0.411 ± 0.019) and IL-22 + PD98059 group (0.280 ± 0.012) than in the 50-μg/L IL-22 group (both P ＜ 0.01).ELISA also showed the same trend of changes in the level of CCL27 in the above groups as Western blot showed.Conclusion IL-22 can promote the expression of CCL27 in HaCaT cells,which may be associated with the MAPK-ERK 1/2 and JAK2/STAT3 signaling pathways.

Objective: To explore the risk factors of occupational exposure of intern nurses in an infectious disease hospital, and put forward prevention and control strategies. Methods: A retrospective analysis was made of 82 reported cases of occupational exposure of intern nurses in a tertiary infectious disease hospital from 2013 to 2017. The occupational exposure rate, pathogens, occurrence sites and exposure links of the intern nurses were analyzed. Results: The overall occupational exposure rate of internship nurses in 2013-2017 was 6.20% (82/1322) .The pathogens were mainly HBV, 52 cases, accounting for 63.4%. The site of occurrence was mainly finger, 75 cases, accounting for 91.5%. In the occurrence of the process, 35 cases were taken, accounting for 42.7%; 20 cases of arteriovenous puncture, accounting for 24.4%; 17 cases of separation needles and finishing materials, accounting for 20.7%. Conclusion The incidence of blood-borne occupation of intern nurses is higher, which needs special attention in infectious disease hospitals. To prevent blood-borne occupational exposure of intern nurses, occupational safety education should be strengthened, clinical operation should be standardized and humanized management should be provided.

BACKGROUNDKCNJ11, ABCC8, PPARG, and HNF4A have been found to be associated with type 2 diabetes in populations with different genetic backgrounds. The aim of this study was to test, in a Chinese Han population from Beijing, whether the genetic variants in these four genes were associated with genetic predisposition to type 2 diabetes.

METHODSWe studied the association of four representative SNPs in KCNJ11, ABCC8, PPARG, and HNF4A by genotyping them using ABI SNaPshot Multiplex System in 400 unrelated type 2 diabetic patients and 400 unrelated normoglycaemic subjects.

RESULTSrs5219 (E23K) in KCNJ11 was associated with genetic susceptibility to type 2 diabetes (OR = 1.400 with 95% CI 1.117 1.755, P = 0.004 under an additive model, OR = 1.652 with 95% CI 1.086 2.513, P = 0.019 under a recessive model, and OR = 1.521 with 95% CI 1.089 2.123, P = 0.014 under a dominant model) after adjusting for sex and body mass index (BMI). We did not find evidence of association for ABCC8 rs1799854, PPARG rs1801282 (Pro12Ala) and HNF4A rs2144908. Genotype-phenotype correlation analysis revealed that rs1799854 in ABCC8 was associated with 2-hour postprandial insulin secretion (P = 0.005) after adjusting for sex, age and BMI. Although no interactions between the four variants on the risk of type 2 diabetes were detected, the multiplicative interaction between PPARG Pro12Ala and HNF4A rs2144908 was found to be associated with 2-hour postprandial insulin (P = 0.004 under an additive model for rs2144908; and P = 0.001 under a dominant model for rs2144908) after adjusting for age, sex and BMI, assuming a dominant model for PPARG Pro12Ala.

CONCLUSIONSOur study replicated the association of rs5219 in KCNJ11 with type 2 diabetes in Chinese Han population in Beijing. And we also observed that ABCC8 as well as the interaction between PPARG and HNF4A may contribute to post-challenge insulin secretion.

ATP-Binding Cassette Transporters ; genetics ; Adult ; Body Mass Index ; Diabetes Mellitus, Type 2 ; genetics ; Female ; Genetic Predisposition to Disease ; genetics ; Genotype ; Hepatocyte Nuclear Factor 4 ; genetics ; Humans ; Male ; Middle Aged ; PPAR gamma ; genetics ; Polymorphism, Single Nucleotide ; genetics ; Potassium Channels, Inwardly Rectifying ; genetics ; Receptors, Drug ; genetics ; Sulfonylurea Receptors

Adipocytes ; cytology ; drug effects ; Animals ; Drug Screening Assays, Antitumor ; methods ; Liver Neoplasms, Experimental ; drug therapy ; Male ; Mice ; Mice, Inbred BALB C ; Mice, Nude ; Rats ; Rats, Sprague-Dawley ; Stromal Cells ; cytology ; drug effects