1.Expression of a testis-specific gene 1700001022RIK in mice and its bioinformatic analysis.
Yu-chi LI ; Shou-ren LIN ; Man-ling LUO ; Huan GUO ; Han-wei WU ; Zhi-mao JIANG ; Yao-ting GUI
National Journal of Andrology 2015;21(5):391-395
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
2.Influence of celecoxib on invasiveness of human high-metastatic nasopharyngeal carcinoma cell line CNE-2Z
Wei-Ren LUO ; Li-Xia LI ; Si-Yi LI ; Han-Guo JIANG ; Xiao-Yi CHEN
Chinese Journal of Otorhinolaryngology Head and Neck Surgery 2010;45(11):941-945
Objective To investigate the effect and mechanism (a selective cyclooxygenase-2 inhibitor) on invasive ability of human nasopharyngeal carcinoma (NPC) line CNE-2Z. Methods The 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). Results MTT showed that celecoxib inhibited CNE-2Z proliferation in dose-dependent manner, the survival rate of cells treated with 25, 50, 100 μ mol/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 ( repectively, 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 Ecadherin 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 ). Conclusion Celecoxib can inhibits the invasive ability of NPC cell line CNE-2Z, which possibly relates with the upregulated expression of E-cadherin.
3.Correlation between perceived hospital ethical climate and body constraint knowledge, attitude and practice among nurses in a Class Ⅲ Garde A hospital
Yan HAN ; Bin HAN ; Shan LUO ; Xuefei REN ; Jiangxia LI
Chinese Journal of Modern Nursing 2020;26(11):1429-1433
Objective:To explore the status quo of nurses' perceived hospital ethical climate (HEC) and body constraint knowledge, attitude and practice, and discuss the correlation between HEC and body constraint knowledge, attitude and practice.Methods:Between April and May 2019, totally 400 clinical registered nurses from a Class Ⅲ Garde A hospital in Shanxi province were selected using convenient sampling, and investigated with the general information questionnaire, Hospital Ethical Climate Survey (HECS) and knowledge, attitude and practice questionnaire. The effects of HEC on nurses' body constraint perception were analyzed. Totally 400 questionnaires were distributed, and 387 valid ones were collected, accounting for an effective recovery rate of 96.75%.Results:The total HECS score of the 387 nurses was (89.72±32.78) . Their scores of body constraint knowledge, attitude and practice were (10.86±2.37) , (30.48±2.94) , and (41.82±7.24) , respectively. There were statistically significant differences in the knowledge, attitude and practice scores between nurses with different educational background, professional title and training experience ( P<0.05) . Correlation analysis revealed that the total score and scores of different HECS dimensions were positively correlated with body constraint knowledge, attitude and practice scores ( P<0.05) . Hierarchical regression analysis showed that the relationships with the doctor, the patient, and the manager in HEC were the influencing factors to nurses' body constraint knowledge and attitude, and the relationships with the doctor and the manager in HECS were the influencing factors to nurses' body constraint practice ( P<0.05) . Conclusions:HEC is closely related to nurses' body constraint knowledge, attitude and practice. Creating a sound HEC will help improve nurses' perception of body constraints, thus standardizing nurses' body constraint practice.
4.Construction and identification of the replication-deficient recombinant vaccinia virus co-expressing HPV type 16 L1 and L2 proteins.
Liqun HAN ; Jiao REN ; Yu LIANG ; Houwen TIAN ; Huijun ZHI ; Weifeng LUO ; Zhenhua LU ; Lanlan WEI ; Li RUAN
Chinese Journal of Experimental and Clinical Virology 2002;16(3):256-260
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
5.Observation on therapeutic effect of electroacupuncture for treatment of sudden hearing loss.
Ren-han LUO ; Jie ZHOU ; Yun-sheng HUANG ; Kai XU
Chinese Acupuncture & Moxibustion 2009;29(3):185-187
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
6.Construction of recombinant vaccinia virus co-expressing mutant E6 plus E7 proteins and detection of its immunogenicity and antitumor response.
Huijun ZHI ; Liqun HAN ; Jiao REN ; Houwen TIAN ; Weifeng LUO ; Yu LIANG ; Li RUAN
Chinese Journal of Experimental and Clinical Virology 2002;16(4):341-344
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
7.Modification of HPV type 16 E6 and E7 genes, and analysis of stability and immunogenicity of the modified proteins.
Huijun ZHI ; Liqun HAN ; Jiao REN ; Houwen TIAN ; Weifing LUO ; Yu LIANG ; Li RUAN
Chinese Journal of Experimental and Clinical Virology 2002;16(2):124-127
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
8. Analysis of blood-borne occupational exposure of intern nurses in a tertiary infectious disease hospital
Jizheng HUANG ; Mingfeng HAN ; Tengda LUO ; Ake REN ; Xiaoping ZHOU
Chinese Journal of Industrial Hygiene and Occupational Diseases 2019;37(7):551-553
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.
9.Effects of interleukin-22 on the expression of CC chemokine ligand 27 in human epidermal keratinocytes
Wenjiao ZHOU ; Jie REN ; Long HAN ; Xinxin LIU ; Kunlong TANG ; Suju LUO
Chinese Journal of Dermatology 2018;51(8):592-594
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.