Objective To analyze the VP1-VP4 genetic region of enterovirus 71 ( EV71 ) strains isolated from children with severe or mild hand, foot and mouth disease ( HFMD) in Shenzhen in 2012. Methods EV71 strains were isolated from five children with mild HFMD and five children with severe HFMD in Shenzhen in 2012.Reverse transcription-polymerase chain reaction ( RT-PCR) method was used to amplify the sequence of VP1-VP4 genes of EV71 strains.The sequences of the amplified products were analyzed by comparing with those of the EV71 reference strains ( A, B and C genotypes) published in Gen-Bank using nucleotide alignment, amino acid alignment and phylogenetic tree analysis.Results The homo-geneity between the EV71 strains isolated from severe and mild cases was 95.1%-98.2% in nucleotides and 99.2%-100% in amino acids.The VP1-VP4 nucleotide sequences of 5 strains isolated from severe cases and 5 strains from mild cases in Shenzhen shared 87.9%-97.8% homologies in nucleotides and 97.3%-99.9% homologies in amino acids with the genotype C EV71 reference strain.The EV71 strains isolated from children in Shenzhen were highly similar with the EV71 strain (FJ439769) isolated in Fuyang in 2008 and the one isolated in Jingdezhen in 2011 (JQ806378, C4a subtype) in nucleotide sequences.Mutations at the residue 31 in the VP1 region ( N→D ) were detected in 3 strains isolated from children with severe HFMD.Conclusion All of the 10 EV71 strains isolated in Shenzhen in 2012 belonged to the sub-genotype C4a.The mutation ( aa31 N→D) in the VP1 region of EV71 might be related to the different clinical mani-festations of HFMD cases in Shenzhen area.

ObjectiveTo investigate the prevalence of 15 skin diseases in adolescents from Liangshan prefecture in Sichuan Province.Methods A stratified cluster random sampling method was used to select subjects.Adolescents from 2 urban and 3 rural middle schools were investigated,who were required to fulfill self-report questionnaires and received physical examinations by doctors.ResultsA total of 7747 adolescents aged from 12 to 20(15.86 ± 1.86) years were included in the investigation,and skin diseases were present in 6379 adolescents,with a total incidence of 82.34％.In average,each subject suffered from 1.3372 ± 0.96861 skin diseases.Of the adolescents,4698(60.64％) had acne,2219(28.64％) freckle,966(12.47％) lichen pilaris,792(10.22％) eczema or dermatitis,511(6.60％) urticaria,195(2.52％) papular urticaria,37 (0.48％) vitiligo,55 (0.71％) psoriasis,166 (2.14％) verruca vulgaris,168 (2.17％) flat wart,317 (4.09％) tinea manus and pedis,142 (1.83％) tinea corporis,62 (0.80％) chloasma,26 (0.34％) nevus flammeus,15 (0.19％) nevus of ota.Male adolescents showed a higher incidence of acne(63.91％ vs.58.50％),but a lower incidence of eczema,lichen pilaris,freckle,chloasma and psoriasis compared with female adolescents.The incidence of acne,vitiligo,psoriasis and melasma were higher,while that of freckele was lower(25.93％ vs.29.58％ ) in Yi adolescents than in Han adolescents.Increased incidence of acne and papular urticaria and decreased prevalence of urticaria,freckle,melasma and lichen pilaris were observed in urban adolescents compared with rural adolescents.ConclusionsThe prevalence of skin diseases is high in adolescents from Liangshan prefecture.Of these 15 skin diseases,acne,freckle,lichen pilaris,eczema and urticaria are the 5 most common diseases in the area.Differences exist in the incidence of skin diseases between male and female,Han and Yi,urban and rural,adolescents.

Objective The aim of this prospective study was to evaluate the changes in the ovarian reverse after myomectomy based on serum anti-Mullerian Hormone (AMH) levels. Methods This is a prospective longitudinal observational study. Serum AMH levels were measured at the baseline and 2 day , 3months after myomectomy in 35 women aging from 36 to 45years.Follicle stimulate hormone(FSH) and luteal hormone(LH) were measured at the same time. 35 women of the same age with fibroid who did not undergo operation were selected as control group. Result (1)AMH level is (1.54 ± 0.95)ng/mL,(1.18 ± 0.77)ng/mL,(1.50 ± 0.58 )ng/mL at 0 day, 2 days and 3 months after operation. AMH level decreased significantly at 2 days after operation (P < 0.05) and increased gradually 3 months after operation, but showed no significant change (P > 0.05).(2) Significant differences in the serial change of AMH levels existed at each time point between myomectomy group and control group (P <0.05). No significant differences in FSH or LH levels existed at each time point. Conclusion AMH is may be superior to FSH or LH in evaluating the changing of ovarian reverse. The study suggests that myomectomy affect the ovarian function for up to 3 months post-operatively , and hemorrhage during and after operation may decrease serum AMH levels.

Objective To evaluate the value of the new ovarian function marker serum anti-Mullerian hormone (AMH)and to clarify the effect of hysterectomy on the ovarian function of the younger women.Methods The serum AMH,follicle stimulating hormone (FSH)and luteinizing hormone (LH)levels in 35 women suffered uterus benign lesion aged 36-45 years and 35 women suffered hysteromyoma without operation were measured at different time.And the ovarian arterial blood flow resistance index (RI ) was measured by Doppler ultrasound. Results Compred with before operation, the serum AMH levels of the patients in hysterectomy group 2 d and 3 months after operation were significantly decreased (P<0.05 or P<0.01).Compared with control group,the serum AMH levels of the patients in hysterectomy group 2 d and 3 months after operation were also decreased (P<0.05).The ovarian arterial blood flow RI of the patients in hysterectomy group 1 and 3 months after operation were increased compared with control group (P<0.05).There were no significant differences in the serum FSH and LH levels in each group between different time points (P>0.05).Conclusion Hysterectomy can affect the ovarian function,and the serum AMH level 3 months after operation is decreased and the ovarian arterial blood flow RI is increased.AMH is superior to FSH or LH in evaluating the changes of ovarian function.

Objective To investigate the epidemiological characteristics of dengue fever in Shenz-hen city in 2014 and to analyze the evolutional characteristics of the epidemic dengue virus(DENV)strains in order to provide scientific guidelines for the prevention and control of dengue fever. Methods Descrip-tive epidemiological analysis was used to analyze the prevalence of dengue fever in Shenzhen city in 2014. Immunochromatography and real-time PCR were performed to detect the specific antibodies(IgM and IgG) and DENV nucleic acids in serum samples collected from suspected cases of dengue fever. Serum samples collected from the patients at early stage of dengue fever were used to infect the C6 / 36 cell line for further isolation of DENV strains. The types of isolated DENV strains were determined by using real-time PCR. E genes of the isolated DENV strains were amplified by RT-PCR and then sequenced. DNAStar and Clustslx (1. 83)softwares were used to analyze the homology between DENV strains isolated in Shenzhen and other areas. A phylogenetic tree based on the sequences of E genes of Shenzhen strains and other sequences of DENV reference strains downloaded from GenBank was constructed for further analysis. Results A total of 454 cases of dengue fever were reported in Shenzhen in 2014 with a male to female ratio of 1. 43 ∶ 1. Local patients accounted for 76. 21% and the rest 23. 79% were imported cases mainly from Southeast Asian and surrounding cities. There were 441 cases reported from September to November,accounting for 97. 14% of all reported cases. Most of the infected subjects were aged 20 to 50,accounting for 76. 73% . Of the 270 samples positive for DENV nucleic acids,strains of DENV-1,DENV-2,DENV-3 and DENV-4 accounted for 87. 41% ,8. 89% ,0. 37% and 2. 22% ,respectively. The phylogenetic tree analysis revealed that the DENV-1 strains belonged to two genotypes,which were genotypeⅠ and genotype Ⅴ. The DENV strains of genotypeⅠ were highly similar to the epidemic strain isolated in Shenzhen in 2010 and the genotype Ⅴstrains were first reported in Shenzhen. The homology analysis of the nucleotides of E genes showed that mi-nor differences in the nucleotide sequences were found between DENV-2 strains. All of the DENV-2 strains belonged to the genotype Ⅳ as indicated by the phylogenic tree. Conclusion There were 454 cases of den-gue fever(including both local and imported cases)reported in Shenzhen city in 2014,reaching an all-time high. DENV-1 was the predominant pathogen in combination with an increased infection of DENV-2. This study indicated that the prevalent DENV strains might be imported from Southeast countries and neighboring cities. Further researches should be conducted to analyze whether dengue fever is endemic in Shenzhen City.

Objective: To understand the awareness level of AIDS knowledge and its influencing factors among middle school students from AIDS high prevalence areas of Yi Autonomous Prefecture (Liangshan Prefecture), and to provide a reference for making propaganda strategies of AIDS knowledge and intervention measures in the relevant departments. Methods: Totally 10 749 students in 4 middle schools were selected from AIDS high prevalence areas of Yi Autonomous Prefecture in Liangshan by the method of cluster sampling. A self-made questionnaire was used to investigate the ways of getting AIDS knowledge and the condition of conducting AIDS life training skills in school. Results: The awareness rate of AIDS-related knowledge among middle school students in Liangshan Prefecture was 44.4%. According to the results of multivariate Logistic regression analysis, female students, junior grade, Yi nationality, poor self-evaluation, lack of parents, living in rural areas, good family economic condition (student self-assessment), living in a community of Yi nationality were all the dangerous factors to the awareness rate of AIDS. School education, radio and television, and health promotion were the main ways for students to acquire AIDS knowledge(88.82%, 80.83%, 73.54%). Minority students, students in severely affected areas received less AIDS life skills training in schools, the time was later, and student needs were not strong. Conclusion The AIDS awareness level of middle school students in Liangshan AIDS high-incidence area is too low, so it is necessary to pay attention to develop targeted programs to strengthen AIDS health education for middle school students.

Objective To explore the correlation of human chorionic gonadotrophin (hCG) level in the follicular fluid on oocyte retrieval day with the number of oocytes retrieved, maturation rate, embryonic development, and pregnancy outcome in controlled ovarian stimulation cycles. Methods The data of 311 IVF/ICSI-ET cycles from 2012 to 2013 was analyzed and stratified according to hCG level in follicular fluid on oocyte retrieval day (<7 nmol/L, 7-14 nmol/L, 14-21 nmol/L, and >21 nmol/L) determined with chemiluminesence method. The number of oocytes retrieved, oocyte maturation rate, fertilization rate, cleavage rate, available embryo rate and pregnancy rate were compared between the groups. Results In the IVF/ICSI-ET cycles, the cycles with hCG level of 14-21 nmol/L in the follicular fluid on the day of oocyte retrieval had significantly higher oocyte maturation rate and fertilization rate than those in the other 3 groups (P<0.05), but the number of oocytes retrieved, cleavage rate, available embryo rate and pregnancy rate, though slightly higher, showed no significant difference from the other 3 groups (P>0.05). In the group with hCG level >21 nmol/L, the oocyte maturation rate and fertilization rate were significantly lower than those in the other 3 groups (P<0.05), and the available embryo rate and pregnancy rate were slightly lower without significant differences from the other 3 groups (P>0.05). Conclusion Follicular fluid hCG level on the day of oocyte retrieval is associated with oocyte maturation, fertilization, embryonic development potential, and IVF outcome. An excessively high follicular fluid hCG level on the day of oocyte retrieval may have negative effects on oocyte maturation and embryo development.

Objective To explore the correlation of human chorionic gonadotrophin (hCG) level in the follicular fluid on oocyte retrieval day with the number of oocytes retrieved, maturation rate, embryonic development, and pregnancy outcome in controlled ovarian stimulation cycles. Methods The data of 311 IVF/ICSI-ET cycles from 2012 to 2013 was analyzed and stratified according to hCG level in follicular fluid on oocyte retrieval day (<7 nmol/L, 7-14 nmol/L, 14-21 nmol/L, and >21 nmol/L) determined with chemiluminesence method. The number of oocytes retrieved, oocyte maturation rate, fertilization rate, cleavage rate, available embryo rate and pregnancy rate were compared between the groups. Results In the IVF/ICSI-ET cycles, the cycles with hCG level of 14-21 nmol/L in the follicular fluid on the day of oocyte retrieval had significantly higher oocyte maturation rate and fertilization rate than those in the other 3 groups (P<0.05), but the number of oocytes retrieved, cleavage rate, available embryo rate and pregnancy rate, though slightly higher, showed no significant difference from the other 3 groups (P>0.05). In the group with hCG level >21 nmol/L, the oocyte maturation rate and fertilization rate were significantly lower than those in the other 3 groups (P<0.05), and the available embryo rate and pregnancy rate were slightly lower without significant differences from the other 3 groups (P>0.05). Conclusion Follicular fluid hCG level on the day of oocyte retrieval is associated with oocyte maturation, fertilization, embryonic development potential, and IVF outcome. An excessively high follicular fluid hCG level on the day of oocyte retrieval may have negative effects on oocyte maturation and embryo development.

OBJECTIVETo obtain 1-4 IgG-like domains of mouse vascular endothelial growth factor receptor 2 (VEGFR2) fusion protein (mVEGFR2D1-4/GST) and identify its antiginicity and biological activity.

METHODSThe gene of mVEGFR2D1-4 was amplified by RT-PCR from 14-days embryos of Balb/c mice. The PCR product was cloned into pET-42a prokaryotic expression vector to construct the recombinant plasmid pET-42a-mVEGFR2D1-4, which was transformed into E. coli BL21 (DE3) strain for mVEGFR2D1-4/GST expression. The fusion protein was identified by SDS-PAGE and Western blotting, and the antigenicity of the protein purified by affinity chromatography was characterized by ELISA. The VEGF blocking effect of the purified protein in human umbilical vein endothelial cells (HUVECs) were evaluated in in vitro cell cultures.

RESULTSThe mVEGFR2D1-4 gene was obtained, which had an identical sequence to that retrieved in GenBank. The prokaryotic expression vector for mVEGFR2D1-4 was successfully constructed as confirmed by enzyme digestion and DNA sequencing. Both Western blotting and ELISA demonstrated the antigenicity of the purified mVEGFR2D1-4 fusion protein, which obviously blocked the effect of VEGF in promoting HUVEC proliferation in vitro.

CONCLUSIONThe mVEGFR2D1-4/GST fusion protein obtained shows a strong antigenicity and biological activity to facilitate further study of active anti-tumor immunotherapy targeting VEGFR2.

Animals ; Cell Proliferation ; Escherichia coli ; genetics ; metabolism ; Female ; Gene Expression ; Genetic Vectors ; Human Umbilical Vein Endothelial Cells ; Humans ; Mice ; Mice, Inbred BALB C ; Plasmids ; Recombinant Fusion Proteins ; genetics ; immunology ; isolation & purification ; Reverse Transcriptase Polymerase Chain Reaction ; Vascular Endothelial Growth Factor Receptor-2 ; genetics ; immunology ; isolation & purification

OBJECTIVETo explore the correlation of human chorionic gonadotrophin (hCG) level in the follicular fluid on oocyte retrieval day with the number of oocytes retrieved, maturation rate, embryonic development, and pregnancy outcome in controlled ovarian stimulation cycles.

METHODSThe data of 311 IVF/ICSI-ET cycles from 2012 to 2013 was analyzed and stratified according to hCG level in follicular fluid on oocyte retrieval day (<7 nmol/L, 7-14 nmol/L, 14-21 nmol/L, and >21 nmol/L) determined with chemiluminescence method. The number of oocytes retrieved, oocyte maturation rate, fertilization rate, cleavage rate, available embryo rate and pregnancy rate were compared between the groups.

RESULTSIn the IVF/ICSI-ET cycles, the cycles with hCG level of 14-21 nmol/L in the follicular fluid on the day of oocyte retrieval had significantly higher oocyte maturation rate and fertilization rate than those in the other 3 groups (P<0.05), but the number of oocytes retrieved, cleavage rate, available embryo rate and pregnancy rate, though slightly higher, showed no significant difference from the other 3 groups (P>0.05). In the group with hCG level >21 nmol/L, the oocyte maturation rate and fertilization rate were significantly lower than those in the other 3 groups (P<0.05), and the available embryo rate and pregnancy rate were slightly lower without significant differences from the other 3 groups (P<0.05).

CONCLUSIONFollicular fluid hCG level on the day of oocyte retrieval is associated with oocyte maturation, fertilization, embryonic development potential, and IVF outcome. An excessively high follicular fluid hCG level on the day of oocyte retrieval may have negative effects on oocyte maturation and embryo development.

Adult ; Chorionic Gonadotropin ; chemistry ; Embryonic Development ; Female ; Fertilization in Vitro ; Follicular Fluid ; chemistry ; Humans ; Oocyte Retrieval ; Oocytes ; physiology ; Pregnancy ; Young Adult