AIM: To establish a HSP90 highly expressing cell line and study the effect of high level of HSP90 on cell stress response. METHODS: The recombined plasimid pSmycHSP, which contains the full length DNA coding for human HSP90?, was introduced into mouse fibroblast cell line NIH-3T3 by electroporation after being subcloned, purified and identified by limited enzyme digestion. Screened by G418, the positive clones were selected and identified by immunofluorescence, immunocytochemistry and Western-blotting. NIH-3T3 cells transfected with empty plasmid served as control, hyperthermia(44 ℃, 20 min, 40 min)was used to simulate oxidative stress. The activity of lactate dehydrogenase (LDH) in supernatant and damage of DNA were detected by automatic biochemistry analyzer and flow cytometer separately to analyze the effect of high-level HSP90 on cell membrane and DNA injuries under stress condition. RESULTS: The rising level of HSP90 was shown by immunofluorescence, immunocytochemistry and Western-blotting in HSP90 overexpressing cell line. There was no difference in the leakage of LDH between HSP90 overexpressing cell line and control, but the damage of DNA was more severe at 44 ℃for 20 min in HSP90 overexpressing cell line than control. Compared with control, the above indices were relieved at 44 ℃ for 40 min in HSP90 overexpressing cell line. CONCLUSION: The NIH-3T3 derived cell line, which stably expressed high level of HSP90, was established. The effect of high-level HSP90 on cells is complex at different intensity of stress, and the protection may be shown at more severe stress circumstance.

AIM: To find the role of heat shock protein 90(HSP90) and tubulin in oxidative stress preconditioning in HepG2 cells.METHODS: The different doses of H2O2 were used to induce cell injury in HepG2 cells.MTT assay,Western blotting and confocal laser microscopy were also used.RESULTS: MTT colorimetry showed that preconditioning(50 mmo1/L H2O2) provided a temporary resistance against subsequent oxidative stress(500 mmol/L H2O2).Western blotting demonstrated that preconditioning increased the levels of HSP90 and tubulin in HepG2 cells,and lessen the declining of HSP90 and tubulin after stress.Tubulin and HSP90's colocalizations in cells with different doses of H2O2 were also observed under laser scanning confocal microscope.CONCLUSION: Tubulin might play important role in oxidative stress preconditioning in HepG2 cells by combining with HSP90.

AIM: To induce preconditioning and oxidative stress by H_2O_2 in HepG2 cells. METHODS: The different doses of H_2O_2 were used to induce apoptosis in HepG2 cells, which was estimated by AO/EB staining, MTT assay and flow cytometry. RESULTS: The different group of HepG2 cells stained with AO/EB showed different staining state. The high dose of H_2O_2 resulted in the increase in apoptosis rate of HepG2 cells and made MTT activity decreased. However, after pretreated with low dose of H_2O_2, the apoptosis rate was decreased and MTT activity was increased. CONCLUSION: The high dose H_2O_2 induced apoptosis in HepG2 cells and the low dose H_2O_2 protected HepG2 cells against the oxidative stress.

Objective:To explore the therapeutic effect of Chinese herbal medicine for prevention and treatment of acute radiation oropharyngeal inflammation of nasopharyngeal carcinoma.Methods:120 cases of nasopharyngeal carcinoma were randomly divided into an experiment group of 60 cases treated with Chinese herbal medicine and a control group of 60 cases with gargling of Dobell's solution.The incidence rate and the extent of radiation oropharyngeal inflammation,total therapeutic time and short-term therapeutic effect in both groups were investigated.Results:There was no oropharyngeal inflammation of grade 0 in both groups. The incidence rate of radiation oropharyngeal inflammation in the experiment group was significantly lower and the effective rate was significantly higher than those in the control group(P0.05).Conclusion:Chinese herbal medicine combined with radiotherapy can relieve acute radiotherapy oropharyngeal inflammation in the patient of nasopharyngeal carcinoma with no significant adverse reaction and do not influence the short-term therapeutic effect.

BACKGROUND: The special genetic law of short tandem repeat on chromosome X (X-STR) makes it incomparable with autosome markers in forensic identification. However, the population genetics data is far less than that of the autosome STR, and especially the haplotype data are rarely reported.OBJECTIVE: To study the genetic polymorphism of 12 X-STR loci in Shenzhen area by pedigree analysis, aiming to provide scientific and effective data for the application of X-STR in forensic medicine and genetics. METHODS: The blood samples of 118 families were taken to extract DNA by Chelex-100, followed by PCR amplification using Investigator Argus X-12 kit. The frequency of alleles of 231 unrelated individuals was counted by direct counting method and Excel software. Hardy-Weinberg equilibrium test was performed on 12 X-STR loci of female samples by chi-square test. Discrimination power and mean exclusion chance were calculated according to the formula. Pedigree analysis was done to identify haplotypes of female samples and the haplotype frequencies of 4 linkage groups in 111 fathers and 119 mothers were calculated using direct counting method and Excel software.RESULTS AND CONCLUSION: In this study, 349 haplotypes were obtained. There were 238, 139, 153 and 157 haplotypes in linkage groups X1-X4, respectively. The polymorphism of DXS10135 locus was the highest with 21 alleles,while the polymorphism of DXS7423 locus was the worst with only 4 alleles. The combined discrimination power was 0.99999999 in males and 0.99999999 in females. The combined mean exclusion chance was 0.99999999 in trio cases,and 0.99999811 in duo cases. These findings indicate that the X-12 detection system has high polymorphism in Shenzhen Han population, and has important application value in forensic individual identification and paternity testing.

Objective To understand the epidemic trend and drug susceptibility of pathogenic bacteria in children with lower re-spiratory tract infection to provide the basis for clinical rational drug use.Methods The sputum culture specimens in these two hospitals during 2013 were performed the pathogenic bacterial culture and the drug susceptibility test respectively.The results were conducted the comparative analysis.Results 6124 strains of bacteria were isolated from the children lower respiratory tract sputum specimens in our hospital during 2013,including 5 121 strains of Gram negative bacteria,accounting for 62.7%,which were domina-ted by Haemophilus influenzae,Klebsiella pneumoniae,Escherichia coli,Bauman Acinetobacter and Pseudomonas aeruginosa;Gram positive bacteria were 2734 strains,accounting for 33.5%,the top two were Streptococcus pneumoniae and Staphylococcus aureus;311 strains of fungi,accounting for 3.8%,and Candida albicans was predominant.1600 strains of bacteria were isolated from the lower respiratory tract sputum specimens in Nanjing Brain Hospital during 2013,including 1 134 strains of Gram negative bacteria, accounting for 70.9%,which were dominated by Klebsiella pneumoniae,Escherichia coli,Pseudomonas aeruginosa and Acinetobact-er Bauman;296 strains of Gram positive bacteria,accounting for 18.5%,Staphylococcus aureus was predominant;170 strains of fungi,accounting for 10.6%,Candida albicans was predominant.The sensitive drugs to Gram-negative bacilli were piperacillin /tazobactam and imipenem,while the sensitive drugs to Gram-positive bacilli were linezolid and vancomycin.Conclusion The Gram negative bacteria are the main pathogens of lower respiratory infection in children,and strengthening bacterial culture and drug re-sistance monitoring is necessary for rational use of antibacterial drugs.

Objective To analyze the characteristics of diagnosis and therapeutic effect of malignant pleural effusion with lung cancer,and explore the prognostic factors and effective diagnosis and treatment plans.Methods A retrospective analysis in-cludes 728 cases of definite pleural effusion with lung cancer from October 2009 to December 2013 in our hospital,which was fol-lowed-up to September 30,2014,and 438 cases were available analyzed.The main outcome measures were incidence,efficiency,pro-gression-free survival and overall survival.Results The overall median progression-free survival and the median survival of malig-nant pleural effusions with lung cancer was respectively 4 months and 8 months.males,small cell lung cancer,massive pleural effu-sion,and right pleural effusion may be the independent factors of local unmanageable malignant pleural effusion by multivariable Lo-gistic regression.The overall survival of pleural effusion with locally control(less than four weeks )was better than those with un-manageable(mOS:9 month vs.5 month,P<0.001).It was no significant difference of the control rate for overall survival prognosis between pleural effusion with two weeks and those with four weeks.histological type,partial remission time,the number of cycles of chemotherapy and TKI therapy may be the independent development risk by progression with Multivariate Cox regression analysis. the progress risk of patients with squamous cell carcinoma and other histological types were lower than that of adeno carcinoma (P=0.007).The progress risk of development of patients with 4 weeks of pleural effusion was significantly lower than that of the non remission (P=0.004),the progress risk of complete chemotherapy cycle number > 2 cycles and takingTKI treatment were significantly reduced (P<0.001;P=0.026).Gender,histological type,pericardial effusion,partial remission time,cycles of chemo-therapy and TKI were the independent prognostic factors for overall survival.The overall survival prognosis of patients with Fe-male,squamous cell carcinoma,no pericardial effusion,and over three cycles of chemotherapy,TKI therapy,and local controlled in 4 weeks was better.Conclusion Male,massive pleural effusion and right pleural effusion are independent predictive factors of local unmanageable malignant pleural effusion.The overall survival of pleural effusion with locally control was better than the patients with refractory control.Histological type,controllable relief time of pleural effusion,cycles of chemotherapy and TKI therapy were the independent predictive factors of progression and overall survival.

Objective:To investigate changes in the circum-maxillary sutures during alternate maxillary expansions and constrictions in a rat model.Methods:Twenty-two male Sprague-Dawley rats (6 weeks old)were used and divided into three groups.In maxillary expansion (ME)group (n=9),an expander was activated for 5 d,and then sacrificed.In alternate maxillary expansions and constrictions (Alt-MEC) group (9 animals),an alternate expansion and constriction protocol (5 d expansion and 5 d constriction for one cycle)was conducted for 2.5 cycles (25 d total),and then sacrificed.The control group com-prised 4 animals with no appliances used,each of two sacrificed on day 5 and day 25 ,respectively.Cir-cum-maxillary sutures (mid-palatal,maxillopalatine,premaxillary,zygomaticotemporal,and frontonasal suture)in each group were characterized histologically.Results:Histological findings of circum-maxil-lary sutures:in control group,the normal sutures were divided into 5 zones,the center was polygon mesenchyme zone,the lateral was mature chondrocyte zone,the next lateral was bone tissue.In ME group,the mid-palatal suture was expanded after ME,the fibre and connective tissue were extracted and invaded into cartilage zone.Osteoblast hyperplasia and new bone formation occurred.The orientation of the new bone trabecula was consistent with force.The osteoclast appeared in some area.Findings of other sutures were similar with those of control group,osteogenesis was active in some area without obvious re-construction in Alt-MEC group,mid-palatal suture and other sutures in some area were widened after Alt-MEC with much more osteoblast hyperplasia and new bone formation.Fibre and connective tissue were extracted,distorted or even broken.Sutures in some areas were narrowed with osteoblast cytopenia and osteoclast hyperplasia.Fibre and connective tissue compressed because of the different force and orienta-tion.Osteoclast count results showed that compared with the control group,the number of the osteoclast was increased only in the palatal suture in ME group (P<0.05 ).Other sutures did not show obvious changes (P>0.05).In Alt-MEC group,the numbers of the osteoclast in circum-maxillary sutures were the most,and had statistical significances (P<0.05 ).Conclusion:These results suggested that cir-cum-maxillary sutures were actively reconstructed after Alt-MEC.while only midpalatal suture had active reaction after ME.

Objective To investigate the analgesia effect and the possible mechanism of intravenous administration with different concentrations of ozonized saline (OS) by observing changes in behavior,plasma tumor necrosis alpha (TNF-α),and interleukin-6 (IL-6) levels after rat plantar incision.Methods Fifty-four health adult male Sprague-Dawley rats were used in the investigation.Twenty four rats were randomly divided into four groups,6 in each group.The rats in group A1 were intravenously administered with 5ml/kg oxygen saline,10min before the incision.The rats in groups B1,C1,and D1 were intravenously administered with 20 μg/ml,30 μg/ml,and 40 μg/ml OS 5 ml/kg.An 1-cm incision was made in the right plantar surface from the heel to the toes according to the method described by Brennan under sevoflurane anesthesia.The 50％ paw withdrawal mechanical threshold (50％ PWMT) and cumulative pain score (CPS) at the times of 24 h before and 2,6,24,48 h after surgery were underwent.Thirty rats were randomly divided into five groups,6 in each group.Groups A2,B2,C2,and D2 processed as well as group 1.All groups except group E2 were made the model of incisional pain.2 ml blood was taken out of the right ventricular 2 h after the operation,then the levels of plasma TNF-α and IL-6 were detected by using enzyme linked immunosorbent assay (ELISA).Results There were no significant differences in the 50％PWMT between group A1 and group B1 at each time point after surgery (P ＞ 0.05).The 50％ PWMT in groups C1 and D1 were higher than those in group A1 at each time point after surgery (P ＜ 0.05).The CPS in groups B1,C1,and D1 were lower than those in group A1 after surgery (P ＜ 0.01).Compared to group E2,the levels of plasma TNF-α 2 h after the operation in group B2 and D2 were not statistically different (P ＞ 0.05).The levels of plasma TNF-α in groups C2 and A2 were higher than those in group E2 (P ＜ 0.05).The levels of plasma IL-6 2 h after the operation between group A2 and group E2 showed no difference (P ＞ 0.05).The levels of plasma IL-6 in groups B2,C2,and D2 were higher than those in group E2 (P ＜ 0.05).Concltsions Intravenous administration of ozonized saline can inhibit the incisional pain in rats.The analgesia effect of ozonized saline was dose-dependent.

Objective To explore the effect of non-invasive labor analgesia on postnatal depression among primipara. Methods Three hundred primipara of spontaneous labor in our hospital from February 2014 to April 2014 were assigned as control group and another 300 primiparas of spontaneous labor from May 2014 to August 2014 as experiment group. In the control group, one-to-one responsibility system was conducted for the delivery with an accompanying midwife and in the experiment group, non-invasive labor analgesia was done apart from one-to-one responsibility system. The two groups were compared in terms of analgesic effect at three stages of labor, postnatal depression scores and incidence of postnatal depression. Result The analgesic effect of the experiment group was better than that of the control group during the three stages of labor (all P<0.01), and postnatal depression scores and the incidence of postnatal depression in the experiment group were significantly lower than those in the control group (both P<0.05). Conclusion Non-invasive labor analgesia can relieve the labor pains and decrease the incidence of postnatal depression among primiparas.