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 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.

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.

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 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.

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 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 study the changes of transforming growth factor-β (TGF-β),connective tissue growth factor (CTGF) and collagen 1 (COL1) in newborn rat's lung tissue and the expressions of 4EBP1 (eukaryotic promoter) and P7OS6K (mammalian target of rapamyein pathway downstream target protein) after rapamycin and hyperoxia intervention,and to study the influence of mammalian target of rapamyein (mTOR) pathway on hyperoxic lung injury and the possible intervention methods.Method A total of 48 21-day-old neonatal rats were assigned into 8 groups (n =6),including air control group,3 d group (3 days after hyperoxic exposure),7 d group (7 days after hyperoxic exposure),14 d group (14 days after hyperoxic exposure),air + RAPA group (air + rapamycin),3 d + RAPA group (3 days after hyperoxic exposure + rapamycin),7 d + RAPA group (7 days after hyperoxic exposure + rapamycin) and 14 d + RAPA group (14 days after hyperoxic exposure + rapamycin).In the hyperoxic group,newborn rats were exposed to 90％ oxygen for 3,7,14 days respectively.The rats in the hyperoxia + rapamycin intervention groups received intraperitoneal injection of rapamycin and inhaled high concentrations of oxygen for 3,7,14 days respectively.Air ± rapamycin group received intraperitoneal injection of rapamycin for 3 days.To study the pathological changes of lung tissures after hyperoxia and rapamycin intervention,we used ELISA to detect the changes of TGF-β,CTGF and COL1 and Western blot to detect the variations of mTORC1,P70S6K and 4EBP1 expression.Result TGF-β,CTGF,COL1 levels at 3 days,7 days and 14 days after hyperoxic exposure (TGF-[β:33.7±2.8 vs.58.6 ±3.1 vs.98.8 ±1.5 ng/mg,CTGF:50.1 ±1.8 vs.68.7 ± 2.2 vs.94.4 ±2.5 ng/mg,COL1:471.9 ±5.7 vs.529.7 ±7.0 vs.556.4 ±8.5 ng/mg) were significantly higher than the air control group (TGF-β:25.5 ± 1.9 ng/mg,CTGF:41.7 ± 1.4 ng/mg,COL1:414.4 ± 8.9 ng/mg) (P ＜ 0.01).While the levels in rapamycin intervention group were significantly lower than all the hyperoxia + rapamycin intervention groups (P ＜ 0.01).The lung tissue pathological grades in 3 d + RAPA group and 7 d + RAPAgroup were significantly lower than those in the 3 d group and 7 d group (3.5 ± 0.8 vs.6.3 ± 2.3 and 9.7 ± 2.0 vs.14.0 ± 2.4) (P ＜ 0.01).The mTORC1,P70S6K,4EBP1 expressions in 3 d + RAPA group were lower than 3 d group (mTORC1:0.26 ± 0.04 vs.0.29±0.08,P70S6K:0.29±0.01 vs.0.31 ±0.08,4EBP1:0.31 ±0.06 vs.0.33 ±0.06) (P＜0.05),while the expressions in 7 d + RAPA and 14 d + RAPA groups were significantly lower than 3 d + RAPAgroup (P ＜0.01).Conclusion mTOR signal pathway may be involved in the repairing process of hyperoxic-induced lung fibrosis.Rapamycin can reduce the levels of TGF-β,CTGF and COL1 and inhibit the expressions of mTOR pathway downstream target protein P70S6K and 4EBP1,thus reduce lung injury atearly stage.