Objective To investigate the prevalence,radiological features and the association between HRCT changes and different diseases or clinical features in interstitial lung disease(ILD)caused by diffuse connective tissue diseases(CTD).Methods Retrospective analysis was performed based on the medical records of 412 in-patients with the diagnosis of diffuse connective tissue diseases from June 2003 to June 2005 in our hospital.268 cases were SLE,83 cases were SS,40 cases were DM/PM,21 cases were SSe.All patients had chest X-ray and the suspected cases had HRCT exam.The distribution of ILD among different diffuse CTD.the appearance between X-ray and HRCT appearances were compared.The correlation among HRCT appearance,different diseases and clinical features was studied.Results①In the 412 cases of CTD,ILD de- tected by HRCT was 9.7%(40 case in total),3,9% by X-ray.Among the 40 cases,DM/PM had the highest rate of ILD(25%),23.8% in SS,9.6% in SS and 6.3% in SLE.②The consensus rate between HRCT and X-ray was 40%.The rate of mis-diagnosis of ILD was 60% by X-ray.③The changes of interstitial involvement de- tected by HRCT were:ground glass change(41.0%),consolidation(32.8%),reticular change(16.4%)and hon- eycomb change(9.8%).The concomitant presence of two of the above changes were very common.④More level respiratory symptoms such as cough,panting,chest distress and chest pain and high level of complement were observed in patients with ground glass and consolidation changes,while high CRP level was seen in patients with consolidation and honeycomb changes(P

Objective: To investigate the incidence trend of sexually transmitted diseases (STDs) in Yantian District, Shenzhen City, Guangdong Province from 2008 to 2022, and the effects of age, period, and cohort on STDs incidence, so as to provide the basis for formulating control measures for STDs. Methods: Data of reported STDs cases (syphilis, gonorrhea, genital Chlamydia trachomatis infections, condyloma acuminatum and genital herpes) among population aged 15 to 64 years in Yantian District from 2008 to 2022 were collected through the Chinese Disease Control and Prevention Information System. The temporal and population distribution of STDs were descriptively analyzed. Trends in incidence of STDs were analyzed using average annual percent change (AAPC). Impacts of age, period and cohort on the incidence of STDs were analyzed by an age-period-cohort model. Results: A total of 6 156 cases of STDs aged 15 to 64 years were reported from 2008 to 2022, and the average annual incidence was 229.92/105. The overall incidence showed an upward trend (AAPC=2.409%, P<0.05), and an upward trend among females (AAPC=5.846%, P<0.05), but the trend was not statistically significant among males (AAPC=0.193%, P>0.05). The incidence of STDs showed an increasing trend followed by a decreasing trend with the increase of age. The incidence trend among males was consistent with the overall population, reaching its peak in the age group of 30 to 34 years (371.04/105), while the incidence among females showed an increasing trend with the increase of age, peaking in the age group of 60 to 64 years (442.15/105). Compared with 2013-2017, the incidence risk of STDs showed an increasing trend in 2018-2022 (RR=1.349, 95%CI: 1.217-1.495). Compared with the birth cohort of 1975-1979, the incidence risk of STDs in 1980-2005 birth cohort showed an increasing trend, with the highest incidence risk in 2000-2004 birth cohort (RR=4.178, 95%CI: 2.717-6.423). Conclusions The overall incidence of STDs among residents aged 15 to 64 years in Yantian District from 2008 to 2022 appeared a tendency towards a rise. Age, period and cohort all had impacts on the incidence, with a greater effect among females than males.

Objective To construct the secretory expression vector of recombinant human C-reactive protein(rhCRP) for its se-cretory expression in Pichia pastoris ,rhCRP was expressed as a secretory protein and purified ,and the immunity reactivity of the purified protein was identified .Methods The DNA fragment of rhCRP which was designed and synthesized was cloned into pPICZαA vector .Recombinant plasmid pPICZαA/rhCRP was linearized by SacⅠand transformed into Pichia pastoris X-33 by elec-trotransformation .The rhCRP was secreted into the medium under the methanol induction .RhCRP was purified by Histamine affin-ity chromatography .The purified rhCRP was identified by SDS-PAGE and Western blotting ,and its immunity reactivity and stabili-ty was identified by indirect ELISA .Results The pPICZαA/rhCRP expression vector was successfully constructed .The rhCRP of 23 × 103 was inducted and successfully expressed as a secretory protein by the recombinant Pichia pastoris strains .The rhCRP was purified by one step up to 90 .42% purity ,and it was showed good immunity and stability by indirect ELISA .Conclusion The rh-CRP with higher purity and immunoreactivity was successfully obtained by using the Pichia pastoris expression system ,which pro-vided an important experimental basis for producing anti-human CRP antibodies and developing testing CRP reagent .

Objective To evaluate the role of autophagy in hippocampal neurons in cognitive dysfunction caused by sevoflurane anesthesia in juvenile rats.Methods One hundred four male SpragueDawley rats, aged 7 days, weighing 10-16 g, were randomly assigned into 3 groups using a random number table: control group (group C, n =8), sevoflurane anesthesia group (group S, n =48), and sevoflurane anesthesia + rapamycin group (group SR, n =48).Group C inhaled 60％ oxygen for 6 h.S and SR groups inhaled 3.6％ sevoflurane anesthesia for 6 h, and in addition, rapamycin 2 mg/kg was injected intraperitoneally at 1 h before of sevoflurane inhalation in group SR.The equal volume of phosphate buffer solution was given in C and S groups.At 1 h before sevoflurane anesthesia (T0) , immediately after the end of anesthesia (T1) , and at 12, 24 and 48 h after the end of anesthesia (T2-4) , 8 rats were randomly selected from S and SR groups to determine the expression of autophagy-related proteins microtubule-associated protein 1 light chain 3 (LC3) Ⅰ , LC3 Ⅱ and Beclin-1 in hippocampal neurons (by Western blot).The ratio of LC3 Ⅱ/LC3 Ⅰ was calculated.Morris water maze test was performed at 5 weeks after the end of anesthesia to assess the cognitive function.The escape latency, frequency of crossing the original platform, and duration of swimming spent at the target quadrant were recorded.Results Compared with the values at T0, the expression of LC3 Ⅱ and Beclin-1 was significantly down-regulated at T1-3 , and the ratio of LC3 Ⅱ/LC3 [was decreased in group S, and the expression of LC3 Ⅱ and Beclin-1 was significantly up-regulated at T1-4, and the ratio of LC3 Ⅱ/LC3 Ⅰ was increased in group SR (P＜0.05).Compared with group C, the escape latency was significantly prolonged, the frequency of crossing the original platform was reduced, and the duration of swimming spent at the target quadrant was shortened at 3-5 days in group S (P＜0.05) , and no significant change was found in the parameters mentioned above in group SR (P＞ 0.05).Compared with group S, the escape latency was significantly shortened, the frequency of crossing the original platform was increased, and the duration of swimming spent at the target quadrant was prolonged at 3-5 days, the expression of LC3 Ⅱ and Beclin-1 was up-regulated at T1-4 , and the ratio of LC3 Ⅱ/LC3Ⅰ was increased in group SR (P＜0.05).Conclusion Weakened autophagy in hippocampal neurons is involved in cognitive dysfunction caused by sevoflurane anesthesia in the juvenile rats.

ObjectiveTo detect the difference of cytokines and antibodies productions by immunologic system from mice and rabbits vaccinated with the M.RCAg-1 chimeric protein,expressed in E.coli,formulation with different adjuvants,including Freund's adjuvant and three clinically acceptable adjuvants,namely,Al(OH)3,Montanide ISA720 and Montanide ISA51.MethodsSix weeks female BALB/c mice were vaccinated with recombinant protein formulated with different adjuvants through intranasal.Serum were collected to detect specific antibodies of M.RCAg-1 and individual Epitope by ELISA ; natural parasite antigen was recognized by indirect immunofluorescence assay; mouse specific T lymphocyte activation was detected by enzyme-linked immunosorbent spot test (ELISPOT) ; Affinity assay between protein and immune IgG of rabbits with the biosensor,and the growth of Plasmodiumfalciparum in vitro to evaluate by growth inhibition assay(GIA).ResultsDifferent formulation can induce different levels of antibody titers,the effection of ISA51 adjuvant was most closely with Freund's adjuvant,and can induce a higher specific antibody of 11 epitopes within proteins,can effectively stimulate cellular immune response based on the IFN-γ,to avidity Montanide ISA51 adjuvant immune antibodies and M.RCAg-1 protein affinity than the other two adjuvants;and Montanide ISA720 adjuvants and Al (OH)3 adjuvant group in mice can't induce a significant IFN-γresponse(P＞0.05).On avidity assay,the Montanide ISA51 formulation group was better than the other two adjuvants; and Montanide ISA720 and Al (OH)3 adjuvant formulation group can't induce a significant IFN-γresponse in mice(P＞0.05) ; the inhibition rates were 60% and 100% in 3D7 and Dd2 Plasmodium falciparum at a concentration of 2 mg/ml IgG by Montanide ISA51 formulated protein,and IgG of Al( OH)3 formulation could not effectively inhibit the in vitro growth of Plasmodium falciparum( 10% ),while IgG of Montanide ISA720 formulation could not inhibit growth of parasite in vitro.ConclusionBy comparing three clinically acceptable adjuvants and Freund's adjuvant in BALB/c mice and New Zealand rabbit,Montanide ISA51 adjuvants can be acceptable formulated M.RCAg-1 protein induced humoral and cellular immune responses,can be used as one of the candidate adjuvants.

ObjectiveTo explore the effects of vector fusion peptides on the conformation and immune reactivity of recombinant polyepitope antigens,M.RCAg-1.MethodsWe subcloned polyepitope artificial antigen gene,M.RCAg-1,into prokaryotic expression vectors,pDS-ex,that contain different fusion tags at the N-terminus or no any tag by different restriction enzyme cutting site.Three recombinant proteins expressed by these vectors,named P312-1,P312-2,and P312-3,were purified and purity is greater than 95％.Then BALB/c mice were vaccinated with the three proteins formulated with Freund's adjuvant through intranasal.Serum were collected to detect specific antibodies of M.RCAg-1 and individual epitope by ELISA ; natural parasite antigen was recognized by indirect immunofluorescence assay; mouse specific T lymphocyte activation was detected by enzyme-linked immunosorbent spot test (ELISPOT) ; and the growth of Plasmodium falciparum in vitro to evaluate by growth inhibition assay(GIA),and analyze secondary and tertiary structures of recombinant proteins from different expression vectors by bioinformatics and circular dichroism technique.ResultsThe P312-1,P312-2 have almost the same amino acid sequence,and the three proteins have the same immunogenicity in animal models(P＞0.05),however,the different proteins elicited various T-cell responses,the rabbit antibody induced by these proteins showed diverse efficacy in malaria parasite growth inhibition assaysin vitro ( respectively,93.9％,14.7％,54.3％ ).The significant differences of secondary and tertiary structures were shown in recombinant proteins from different expression vectors,analyzed by bioinformatics and circular dichroism technique,which demonstrated the change of protein molecule spaces conformation,and the obviously change of some epitope locations.ConclusionThese results suggest that the expressed polyepitope artificial antigens originating from the different vector fusion peptides indeed affect the protein folding and,subsequently,the epitope exposure.Thus,these proteins are able to induce both distinct humoral and cellular immune responses in animal models,and they affect the efficacy of immune inhibition against the parasite,the enhancement or suppresses.

Objective: To investigate the differentially expressed genes in osteosarcoma cell lines with various metastatic potentialities, and to screen for new candidate genes related to metastasis of osteosarcomas. Methods: The total RNAs of a lowly metastatic and a highly metastatic osteosarcoma cell lines (M6 and M8) were extracted. Differentially expressed genes in the two osteosarcoma cell lines were studied by cDNA microarray. The hybridization signals were scanned with a Generation Ⅲ array scanner and analyzed by Imagequant 5.0 software. Typical differentially expressed genes were further verified by real-time quantitative PCR. Results: There were 330 differentially expressed genes between M6 and M8 cells. In the high-metastasis M8 cells, 178 genes were up-regulated and 152 genes were down-regulated compared to the low-metastasis M6 cells, with 43 extremely up-regulated and 49 extremely down-regulated. The differentially expressed genes were mainly associated with cell proliferation, indicating these genes might be related to the inhibition of M6 cells. Other differentially expressed genes included those associated with the regulation of gene expression and signal transduction, indicating these genes might be correlated with tumor metastasis. Conclusion: cDNA microarray shows an advantage in identifying genes associated with metastasis of osteosarcoma. In M8 subset of MG63 osteosarcoma cells,43 genes are up-regulated and 49 genes are down-regulated, which may be related with metastasis of osteosarcoma.

Objective Virus-specific RNA interference (RNAi) is a powerful inhibitor of gene expression and replication of HBV. It is known to have high efficiency, specificity, and few side effects. We wanted to evaluate the effects of siRNA silencing HBV replication on the growth of hepatocellular carcinomatic(HCC) cells to find out an ideal method for treatment of HCC. Methods We transfected siRNA into HepG2.2. 15 cells (HCC cell inserting HBV gene) and detected the HBsAg and HBV DNA copies for evaluating the inhibitory effects of siRNA. Then we evaluated cell growth and self-renewal ability after transfection of siRNA by MTT. Results The HBsAg level and HBV DNA copies were reduced after the transfection of siRNA, the highest inhibition rate was 83.9%,while the inhibition rate of HBV DNA copies reached 73. 4%. The siRNA group's growth ability and self-renewal rate were lower than the control group in 5 days. Conclusion siRNA can effectively inhibit HBV replication and expression in HepG2.2.15 cells and silencing HBV replication can inhibit HepG2.2.15 cell's growth and self-renewal.

Objective: To investigate the effects of hyperbaric oxygen(HB0_2) on black pigmented bacteroides groups(BPB), obligately and facultatively anaerobes of periodontitis. Methods: 60 periodontitis cases were divided into 2 groups. The HB0_2 group(gingival scaling was done at the first visit)was exposed in 0.25 MPa pure oxygen 90 minutes each day, and treated 10 days continuously. The control group received gingival scaling at the first visit. The clinical indices of each group were measured. The microorganisms in periodontal pocket were incubated and analyzed. The number of subgingival anaerobes as well as the number of BPB,obligately and facultatively anaerobes were measured and counted by routine anaerobic culture. Results: Highly significant differences in Gingival Indices (GI), Probing Depth (PD), Attachment Loss (AL), and also significant differences in Plaque Index (PLI),and teeth mobility were seen between the HB0_2 group and the control group. There were significant difference of PLI, GI,PD,AL, teeth mobility in HB0_2 group pre-therapy and post-therapy. There were significant difference of subgingival anaerobes amounts, obligately and facultatively anaerobes sorts between pre-therapy and post-therapy. The positive rate of BPB of group HB0_2 post-therapy were reduced more than that of pre-therapy(P<0.01). Conclusion: HB0_2 has beneficial therapeutic effects on chronic and aggressive periodontitis. The mechanism of the HB0_2 therapy might inhibit the growth of subgingival obligately and facultatively anaerobes and BPB of human periodontitis, especially the obligately anaerobes and BPB in periodontal pockets.

Objective: Asphyxia and ventricular fibrillation are the two most prevalent causes of cardiac arrest. The study investigated the differences in brain damage after cardiac arrest between asphyxial and ventricular fibrillation cardiac arrests in rats. Methods: Male healthy Sprague-Dawley rats were randomly assigned to the asphyxial group (cardiac arrest of 6 min, n=15), ventricular fibrillation group (cardiac arrest of 6 min, n=15) and sham group (n=5). Neurologic deficit scores and tape removal test were evaluated at 1, 3 and 7 days after cardiopulmonary resuscitation from three groups. Serum S-100B and brain histopathologic damage scores were also examined. Results: There were no differences in neurologic performance at 1, 3 and 7 days after cardiopulmonary resuscitation between the asphyxial group and ventricular fibrillation group (P>0.05, respectively). Serum S-100B level was higher in the asphyxial group at 1, 3 and 7 days, compared with the ventricular fibrillation group (P<0.05, respectively). There were significantly higher histopathologic damage scores at 1, 3 and 7 days in the asphyxial group compared with the ventricular fibrillation group (P<0.05, respectively). Conclusion: Asphyxial cardiac arrest has worse morphologic brain damage compared with ventricular fibrillation cardiac arrest, but the functional brain damage caused by asphyxial cardiac arrest is similar to that caused by ventricular fibrillation cardiac arrest.