Abstract: Objective　To analyze the epidemiological characteristics of late diagnosed HIV/AIDS cases (LD) in Sanya from 2010 to 2021, and to provide evidence for reducing the LD rate. Methods　The database was downloaded from the AIDS Prevention and Control Information System of China's Disease Prevention and Control Information System and newly reported HIV/AIDS cases between 2010 and 2021 in Sanya were included, identified LD according to the LD criteria proposed by Chinese Center for Disease Control and Prevention in 2014 and analyzed the relevant factors of LD. Results　From 2010 to 2021, a total of 710 research objects were included in this study. The proportion of LD was 33.4% (237/710), and decreased from 95.5% to 22.4% between 2010 and 2021 (χ2trend=34.777, P<0.001). Ethnic groups, educational level, sample sources and confirmed date were the relevant factors of LD of HIV/AIDS in Sanya City. The proportion of LD was 56.8% in Li ethnic group, which was higher than that in Han ethnic (OR=2.253, 95%CI=1.361-3.670). The proportion of LD of patients who were middle school and less was 55.5%, which were more likely to be LD than high school or above (OR=1.722, 95%CI=1.072-2.765). The proportion of LD was 56.8% in patients whose samples were from medical institutions or testing consultation were more likely to be LD than MSM (OR=5.564, 95%CI=3.278-9.444; OR=2.204, 95%CI=1.239-3.923). Compared with patients who were confirmed between 2018-2021, the patients derived from 2010 to 2013 had higher LD (OR=2.246, 95%CI=1.311-4.488). Conclusion　The LD of HIV/AIDS in Sanya cannot be ignored, especially the HIV/AIDS from counseling and testing and medical institutions. We should strengthen HIV testing, strengthen health education.

Objective:To investigate the molecular interaction between non-structural protein 3 serine protease of hepatitis C virus(HCV)and wild type P53,and to lay the basis for elucidating the mechanism of oncogenesis of hepatocellular carcinoma(HCC)after infection of HCV.Methods:The recombinant plasmids,pGAD424-NS3,pGAD424-NS315aa- and pGAD424-NS330aa-,were constructed and the interaction between NS3 serine protease and its cofactor NS4A,the interaction between wild type P53 and NS3 serine protease and its N-truncated mutants were dectected qualitatively and quantitatively in yeast two-hybrid system.Results:The results indicated that interaction existed not only between full-length NS3 serine protease and P53,but also between N-truncated mutants of NS3 serine protease and P53.Furthermore,the difference between enzyme activity unit(IU)of β-gal induced by these interactions was not significant(P>0.05).Conclusions:NS3 serine protease of hepatitis C virus and its N-truncated mutants can interact with wild type P53,and the region of NS3 serine protease involved in the interaction may be located in its C-terminal,but not in its N-terminal.

OBJECTIVETo explore the effect of Yijing Huoxue Cuyun Decoction (YHCD) in adjusting hypoestrogenemic response induced by clomiphene.

METHODSInfertile patients caused by ovulation disturbance were randomly divided into 2 groups. The 60 patients in the observed group were treated with clomiphine plus YHCD, and the 58 patients in the control group were given clomiphine plus estradiol valerate.

RESULTSBy scoring on the cervical relaxation and improvement of cervical mucus, 38 patients (63.3%) in the observed group had Insler score of more than 8 points, while that in the control group was only 25 (43.1%), comparison between the two groups showed significant difference (P < 0.05). The endometrium thickening in the observed group was 0.98 +/- 0.19 cm, significantly different to that in the control group (0.85 +/- 0.21 cm, P < 0.01). Twenty-five patients in the observed group (41.7%), and fourteen patients in the control group (24.1%), respectively got pregnancy, the pregnant rate in the former was obviously higher than that in the latter (P < 0.05).

CONCLUSIONYHCD can ameliorate hypoestrinemia induced by clomiphene and increase the pregnant rate in patients.

Adult ; Clomiphene ; adverse effects ; therapeutic use ; Drugs, Chinese Herbal ; therapeutic use ; Estrogens ; blood ; deficiency ; Female ; Humans ; Infertility, Female ; drug therapy ; Ovulation ; drug effects ; Ovulation Induction ; Phytotherapy

OBJECTIVEElectron microscopical study of infected cells to identify the pathogenic agent of SARS.

METHODSVero E6 cells infected with lung autopsy samples or nasopharyngeal swabs from SARS patients of Beijing and Guangzhou were inoculated. The supernatant and cultured cells exhibiting identifiable cytopathic effect (CPE) were prepared for electron microscopic study.

RESULTSExamination of CPE cells on thin-section revealed characteristic coronavirus particles within the cisternae of endoplasmic reticulum, Golgi apparatus, vesicles and extracellular space. They were mainly spherical or oval in shape, annular or dense, about 80 nm in diameter. Negative-stain electron microscopy identified coronavirus particles in culture supernatant, 80 - 120 nm in diameter, with club-shaped surface projections. Elongated, rod-, kidney- or other irregular shaped virons with the size of 100 - 200 nm by 60 - 90 nm were also found in the cultured cells infected with the lung samples from the Guangdong patients. Infectious virons entered cells by endocytosis or membrane fusion and released through a budding process.

CONCLUSIONThese data indicate a novel coronavirus as the causative agent of SARS. Most viral particles showed typical characteristics of coronavirus. The potential role of special shape viruses is expected to be further investigated.

Animals ; Cercopithecus aethiops ; Humans ; Microscopy, Electron ; SARS Virus ; ultrastructure ; Severe Acute Respiratory Syndrome ; virology ; Vero Cells

Objective：To investigate the correct expression of dengue 2 virus 43 strain NS1 gene in transfected BHK-21 cell. Methods：The D2-43 DNA fragment coding for signal peptide plus NS1 protein was cloned between KpnⅠ site and EcoR Ⅰ site of expression plamid pcDNA3.1. The obtained recombinant vector pcDNA-NS1 was transfected into BHK-21 cells with electroporation technique. After selection by G418, resistant clones were screened by RT-PCR and Western blotting test. Results：The RT-PCR results of four in five randomly selected cell clones were positive. Western blotting test showed that NS1 gene could be expressed in BHK-21 cells. Conclusions：NS1 protein was capable of being expressed and appropriately processed in pcDNA-NS1 transfected BHK-21 cells. The present results suggest the feasibility of NS1-based DNA immunization.

OBJECTIVETo explore the functions of phospholipase C (PLC)-independent protein kinase C signaling pathway (PTH/nonPLC/PKC) of parathyroid hormone (PTH) and its role in bone metabolism.

METHODSOsteoblasts isolated from the calvaria of 2- or 3-day-old C57BL mice, identified by alkaline phosphatase staining and Alizarin red staining, were treated for 4 h with 100 nmol/L [Gly(1), Arg(19)]hPTH(1-28) plus 10 nmol/L RP-cAMP, 10 nmol/L [Gly(1), Arg(19)]hPTH(1-34) plus 10 nmol/L RP-cAMP , 10 nmol/L PTH(1-34), or and 0.1% trifluoroacetic acid (TFA). The total RNA was then isolated for screening differentially expressed genes related to PTH/nonPLC/PKC pathway using Affymetrix mouse 12x135K gene expression profile microarray, and the identified genes were confirmed by real-time quantitative PCR. MC3T3-E1 cells treated with [Gly(1), Arg(19)]hPTH(1-28)+RP-cAMP, [Gly(1), Arg(19)]hPTH(1-34)+RP-cAMP, [Gly(1), Arg(19)]hPTH(1-34)+ RP-cAMP +100 nmol/L Go6983, or 0.1% TFA were also examined for GR(1-28)- or GR(1-34)-mediated gene expression changes using real-time quantitative PCR.

RESULTSAlizarin red staining visualized red mineralized nodules in the osteoblasts at 28 days of culture. According to the genechip results, we selected 56 target genes related to PTH/nonPLC/PKC pathway, among which CITED1 showed higher expressions in [Gly(1), Arg(19)]hPTH(1-34)+ RP-cAMP group than in both the control group and [Gly(1), Arg(19)]hPTH(1-28)+RP-cAMP group (P<0.05), and its expression was the highest in PTH(1-34) group (P<0.05). RT-PCR of MC3T3-E1 cells yielded consist results with those in the primary osteoblasts, and the cells treated with Go6983 (a PKC inhibitor) did not show GR(1-28)- or GR(1-34)-mediated differential expression of CITED1.

CONCLUSIONThe activation of PLC-independent protein kinase C signaling pathway of PTH enhances the expression of CITED1 in mouse osteoblasts to mediate the effect of PTH on bone metabolism, and this pathway is not dependent on the activation of PLC or PKA signaling.

Animals ; Cells, Cultured ; Indoles ; Maleimides ; Mice ; Mice, Inbred C57BL ; Nuclear Proteins ; physiology ; Osteoblasts ; physiology ; Parathyroid Hormone ; physiology ; Protein Kinase C ; physiology ; Signal Transduction ; Skull ; Trans-Activators ; physiology ; Type C Phospholipases

OBJECTIVETo generate rescued viruses with deletion mutation of capsid protein from dengue virus type 2 isolated in China (DEN2-43).

METHODSOn the basis of infectious full-length cDNA clone pD212 of DEN2-43 strain virus, the deletion mutants were constructed by fusion PCR, from which the rescued viruses with deletion mutation of capsid protein were generated by transcription in vitro and electroporation.

RESULT AND CONCLUSIONSequence analysis demonstrated that the deletion mutations had been successfully inserted into the rescued viruses obtained. These mutant viruses may hold the key for elucidating the effects of deletion mutation of capsid protein on the biological characteristics of dengue virus.

Amino Acid Sequence ; Animals ; Base Sequence ; Capsid Proteins ; genetics ; Cell Line ; Cloning, Molecular ; DNA Mutational Analysis ; Dengue Virus ; genetics ; isolation & purification ; Electroporation ; Molecular Sequence Data ; Sequence Deletion ; Sequence Homology, Nucleic Acid ; Transcription, Genetic ; Virus Replication ; genetics

OBJECTIVETo investigate the effect of the non-PLC-dependent protein kinase C (PKC) pathway of parathyroid hormone (PTH) on the apoptosis and proliferation of osteoblast MC-3T3E1 cells.

METHODSMC-3T3E1 cells were seeded in 96-well plates at the density of 1.5×10(4) cells/mL and incubated for 3 day. The cells were then exposed to 100 nmol/L of [Gly(1), Arg(19)]hPTH(1-28), 100 nmol/L of [Gly(1), Arg(19)]hPTH(1-34), 100 nmol/L of [Gly(1), Arg(19)]hPTH(1-34)+1 µmol/L Go6983, 1 µmol/L Go6983, or deionized water (control) for 1, 24 or 48 h. After the treatments, cell counting kit-8 (CCK-8) and Caspase-Glo® 3/7 Assay (Caspase-3) were used to examine the proliferation and apoptosis of MC3T3-E1 cells.

RESULTSCCK-8 results showed that hPTH(1-34) increased the number of MC3T3-E1 cells compared with hPTH(1-34)+Go6983 at 1 h and 24 h, but this difference was not statistically different. At 48 h, treatment with hPTH(1-34), as compared with hPTH(1-28), significantly increased the number of MC3T3-E1 cells (P<0.05), and this effect was blocked by the PKC inhibitor Go6983 (P<0.05). hPTH(1-34) did not result in significant inhibition of MC3T3-E1 cell apoptosis at 1 h and 24 h as compared with hPTH(1-34)+Go6983, but significantly inhibited the cell apoptosis as compared with hPTH(1-28) (P<0.05); this inhibitory effect was blocked by Go6983 (P<0.05).

CONCLUSIONs A relatively long time (for 48 h) of exposure to PTH can inhibit apoptosis and promote the proliferation of MC3T3-E1cells through a non-PLC-dependent PKC pathway.

3T3 Cells ; Animals ; Apoptosis ; Cell Proliferation ; Indoles ; pharmacology ; Maleimides ; pharmacology ; Mice ; Osteoblasts ; Parathyroid Hormone ; pharmacology ; Protein Kinase C ; antagonists & inhibitors ; metabolism ; Signal Transduction

OBJECTIVEX-linked inhibitor of apoptosis protein (XIAP) To gastrointestinal (GI) investigate the expression of XAF1 and heat-shock transcription factors 1 èHSF1éand their relationship in human gastrointestinal cancers.

METHODSImmunoblotting was used to analyze the expression of HSF1 and XAF1 in either gastric or colon cancer tissue and GI cancer cell line. Transient expression of the HSF1-containing vector in GI cell lines and RNA interference were used to up/down-regulae the expression of the HSF1, and the subsequent expression of XAF1 was measured.

RESULTSThe expression of HSF1 was higher in GI cancers than in normal tissues. The expression of XAF1 and HSF1 was negatively correlated in GI cancer cell lines. Stress stimuli up-regulated the expression of HSF1 while the alteration of XAF1 expression was negatively correlated with HSF1 expression.

CONCLUSIONThe high expression of HSF1 in GI cancers is associated with suppressed expression of XAF1, which can be one of the mechanisms for low-expression of XAF1 and insufficient apoptosis in GI cancers.

Animals ; Base Sequence ; Cell Line, Tumor ; DNA-Binding Proteins ; genetics ; Gastrointestinal Neoplasms ; genetics ; metabolism ; pathology ; Gene Expression Regulation, Neoplastic ; Heat Shock Transcription Factors ; Humans ; Intracellular Signaling Peptides and Proteins ; Neoplasm Proteins ; genetics ; Oxidative Stress ; genetics ; Temperature ; Transcription Factors ; genetics

We report a case of ovarian function fluctuation during long-term follow-up in a patient with premature ovarian insufficiency (POI). The patient finally obtained clinical pregnancy with subsequent uneventful full-term delivery after several intracytoplasmic sperm injection-embryo transfer (ICSI-ET) cycles. This case demonstrates that hormone replacement therapy (HRT) and assisted reproductive therapy should be applied as soon as possible to young patients with POI who have a strong desire for pregnancy in the absence of contraindications. This strategy helps such patients obtain pregnancy and delivery before the exhaustion of ovarian function.