ObjectiveTo explore the relationship between the mRNA and protein expressions of NIX and the pulmonary alveolus apoptosis following severe acute lung injury (ALI).Methods Rat models of severe collision injury on the chest were built.The mRNA and protein expressions of NIX in the alveolar cells at 6,12,24,48,72 and 96 hours after injury were detected using immunohistochemistry,immunoblotting and RT-PCR.Meanwhile,apoptosis of the alveolar cells was checked at different time points with Tunel assay.ResultsThe protein expression of NIX in the alveolar cells was observed both in experimental and control groups,which increased at 6 h post injury,peaked at 48 h and then declined till approaching the pre-injury level at 96 h.In the meantime,NIX showed a high expression both in the vascular endothelial cells (VECs) and the renal interstitial fibroblasts.The apoptosis of alveolar cells mainly presented in bronchi,blood vessel endothelium (BVE) and alveolar epithelium at 24 h post injury.The post-injury apoptosis rate of the alveolar cells was significantly higher than the pre-injury rate ( P ＜ 0.01 ),which reached the peak at 72 h and then decreased gradually.The changes of NIX protein in lung tissue showed a positive correlation with the apoptosis rate of alveolar cells after injury (r =0.303,P ＜ 0.01 ).ConclusionsThe up-regulated expression of NIX takes part in the pathophysiological process of apoptosis of the alveolar cells and shows consistency with the apoptosis rate change of the alveolar cells,as may be the molecular basis for apoptosis of the alveolar cells after ALI.

Objective To study the expression of IL-6 and IL-8 in the lung injuries induced by blunt chest trauma under the closure and open state of glottis.Methods The expressions of IL-6 and IL-8 mRNA were detected by RT-PCR in injuried lung tis-sues under the closed and open glottis states snd the control lung tissues at designed different time points.Results The expressions of IL-6 and IL-8 mRNA at 4,8,12h in the closed glottis group were obviously higher than those in the open glttis group (P <0.05) and the expressions at designed various time points in the injured lung tissue were higher than those in the control lung tissues (P <0.05).The change rule of IL-6 and IL-8 mRNA expression in the control group,closed glottis group and open glottis group was similar,the expression was up-regulated.Conclusion IL-6 and IL-8 might play an important role in the occurrence and develop-ment of functional lesion in the injured lung by blunt chest trauma.

ObjectiveTo observe the effect and safety of postoperative patient-controlled epidural analgesia with dezocine plus ropivacaine in patients underwent hysterectomy.Methods60 patients,ASA Ⅰ ～ Ⅱ,undergoing hysterectomy operation.were randomly divided into two groups.0.225％ ropivacaine with 0.005％ dezocine ( DR group) or 0.225 ％ ropivacaine with 0.003％ morphine (MR group) were given epidurally after surgery,respectively,with an initial loading dose 5m l,basal dose of 2ml/h,PCA dose of 0.5ml and lockout time of 15 min.Visual analogue score(VAS),Bruggemann comfort scale(BCS),Ramsay sedation score(RSS),PCEA effective compression of the times and adverse effects were determined and compared at 1 h,4h,8h,12h,24h 36h and 48h after operation.PCEA effective compression of the number and incidence of adverse reactions were observed.ResultsThere were no significant differences in VAS,BCS,RSS and PCEA effective compression of the times between two groups( all P ＞ 0.05 ).The percentage of nausea,vomiting and pruritus were significantly lower in group DR than that in group MR( all P ＜0.05).ConclusionPatient-controlled epidural analgesia with dezocine plus ropivacaine in patients underwent hysterectomy had analgesic effect and safety,with less advers eeffects.

Objective To explore the effect of CYP2D6 gene polymorphism on blood concentration,therapeutic efficacy and adverse effects of anti-depression drug venlafaxine.Methods 69 cases of patients with depression were selected randomly from southern region of FuJian,China.The blood concentration of venlafaxine was detected by HPLC,and the CYP2D6 genotype was determined by sequencing with the amplified PCR products from peripheral blood DNA.The therapeutic efficacy and adverse effects of venlafaxine were evaluated by Hamilton depression scale(HAMD) and treatment emergent symptom scale(TESS) respectively.Results Subjects was divided into CC,CT,TT three groups based on rs16947 locus genotyping.The blood concentrations of venlafaxine were (157.35±15.63) ng/ml,(70.17±5.11) ng/ml,(115.72± 10.2) ng/ml respectively and there was no significant difference (F=1.257,P=0.301).The dose-corrected concentrations and normalized concentrations were not significantly different (F=1.683,1.547,P＞ 0.05).Similarly,the reduction rate of HAMD (CC:(40.6 ± 7.23) ％,CT:(51.7±7.09)％,TT:(42.8±14.1)％) and TESS scores (CC:1.3±0.21,CT:1.3±0.36,TT:1.2±0.28) were not significantly different either (P＞0.05).In 69 samples in this study,genotyping of rs3892097 locus only found GG type,and no further examination was performed.Genotyping divided rs1065852 locus into CC,CT,TT three groups.The blood concentrations of venlafaxine were (42.87±9.9) ng/ml,(64.25 ± 13.59) ng/ml,(181.56± 14.15) ng/ml respectively,and there was significant difference among the three groups (F=4.893,P=0.016).The dose-corrected concentrations and normalized concentrations were also significantly different (F=3.985,3.648,P＜0.05).The reduction rate of HAMD (CC:(42.6±8.23) ％,CT:(48.8± 10.8) ％,TT:(63.4±9.15) ％) was not significantly different (F=2.961,P=0.07).The difference of TESS scores was similar to that of HAMD reduction rate.Conclusion Among the 3 loci studied,only rs1065852 locus within CYP2D6 gene can affect its enzyme activity and then influence the therapeutic efficacy and adverse effects of venlafaxine.

Objective To detect biofilm formation and biofilm-associated genes of methicillin-resistant Staphylococcus aureus (MRSA) in clinical isolates. Methods The biofilm were determined by microtiter plate assay (MPA) and congo red agar (CRA) and the biofilm-associated genes icaA,sarA,fnbA,fnbB were detected by PCR in 33 strains of MRSA in clinical isolates. Results Of the 33 MRSA isolates, 29(87.9%) were MPA positive, 16(48.5%) were CRA positive; The icaA gene was present in 39.4% of isolated strains. Furthermore, 69.7% of strains harboured the sarA gene, 39.4% were fnbA positive and 75.8% were fnbB positive. As many as 87.9% strains had the ability to form biofilm in vitro. 44.8% of MRSA formed biofilm in ica-dependent mechanism and 55.2% of MRSA isolates formed biofilm in ica-independent mechanism. Of the biofilm positive MRSA, 75.9% were sarA positive, 37.9% were fnbA positive and 79.3% were fnbB positive. Conclusion Most of the MRSA strains formed biofilm in ica-independent mechanism. fnbB and sarA gene shows higher frequency among the biofilm-associated genes of MRSA, it may infer that most of the MRSA strains biofilm formation are fnbB-mediated. Meanwhile, sarA may be a positive regulator of fnbB, and thus drives the biofilm formation.

Objective To investigate the lipoteichoic acid(LTA) induced apoptosis and the expression of inflammatory cytokines in human alveolar macrophage (AM) and the anti-apoptotic and anti-inflamatory effect of moxifloxacin (MXF).Methods Obtained human AM from bronchoalveolar lavage and used MTT assay to observe the effects of LTA and MXF on cell activity,optical microscope to investigate the change of the cell morphology,flow cytometry to assess cell apoptosis,RT-PCR to detect the mRNA levels of TLR2,IL-1 β,IL-8 and TNF-α,ELISA for the production of IL-8 to exam RT-PCR.Results LTA showed cytotoxicity on AM in a dose-dependent manner ( P＜0.05 ) ; MXF inhibited the effect of LTA without cytotoxicicy ( P＜0.05 ).LTA promoted apoptosis ( P＜0.05 ) and the mRNA expressions of TRL2,IL-1 β,IL-8 and TNF-α significantly in AM (P＜0.05),the peaks and peak time ofthe above factors were (3.56±0.03) at 12 h,(46.63±7.06) at 6 h,(28.07±1.24) at 12 h and (2.34 ±0.50) at 3 h respectively and increased the release of IL-8 protein level at 24 h (P＜0.05).MXF inhibited the cell apoptosis and the above mRNA expression at 12h ( P＜0.05 ),and inhibited the IL-8 protein level at 24 h( P＜0.05 ).Conclusion LTA showed cytotoxicity on AM,induced AM apoptosis and increased the expression of TLR2,IL-I β,IL-8 and TNF-α of AM ; MXF could protect AM through inhibiting of the above effects and may play a key role beside bactericidal effect in gram-positive bacteria pneumonia.

AIM:Panton-Valentine leukocidin(PVL)is a pore-forming toxin secreted by Staphylococcus aureus epidemiologieally associated with the often-lethal necrotizing pneumonia.Until now,the mechanisms of pathogene-sis of PVL leading to the fatal pulmonia remains undefined and also acquired plenty of the toxins is difficult.In the present study，we obtain recombinant staphylococcal F and S components of the Panton-Valentine leukocidin by gene engineering and evaluate its biological activity in vitro,which provides an experimental basis for the further studies of its biological func-tion and its toxicity in pneumonia.METHODS:The full-length of F and S components of PVL gene amplified from the strain of Staphylococcus aureus DNA by hiSh-fidelity PCR was cloned into prokaryotic expression vector pET22b(+),and the vector was transformed into BL21(DE3)plysS to construct a prokaryotie expression system.The integrity of the opening-reading frame of each construct was verified by DNA sequencing.The recombinant PVL(rPVL)was induced by1.0 mmol/L IPTG.The expressed products were identified by SDS-PAGE and the fusion proteins(6His-LukS-PV and 6His-LukF-PV)were purified from lysates of transfeeted E.coli cells by affinity chromatography on nitrilotriacetic acid columns.The eytolytie activity was tested by incubation of rPVL with human polymorphonuclear neutrophils(PMNs)in vitro.RESULTS:The nueleotide sequence of the cloned PVL gene was the same as that of reported in GenBank.E coli BL21(DE3)plysS containing recombinant vectors grow at 37℃causes some proteins to accumulate as inclusion bodies.while incubation at 30℃led to a significant amount of soluble active proteins which accounted for about 31.7％ of the total bacterial protein.The relative molecular weight showed on SDS-PAGE profile was consistent with the expected value which the LukS-PV protein was about 34 kD.and the LukF-PV protein was about 35 kD.The purified rPVL was obtained and its cytolytic activity to PMNs was demonstrated.CONCLUSION:The genes of 1ukS-PV and lukF-PV are successfully cloned into plasmid pET22b(+)and expressed in E coli respectively.which provide a basis for analyzing the toxicity related to the diseases and further studies about the pathogenesis of PVL.

Objective To characterize hypersensitivity reactions to chemotherapy with carboplatin in patients with gynecologic malignancies and serve use of carboplatin.Methods We retrospectively analyzed the clinical features,management,or outcome of carboplatin-related hypersensitivity reactions in 13 patients with gynecologic malignancies from 1983 to 2008.Results Twenty times hypersensitivity reactions happened in thirteen women with carboplatin hypersensitivity reactions.The earliest one was at the 5th cycle,the last one was at the 28th cycle;the average cycle was 11.6.The accumulative dosage of carboplatin was 1 900-11 400 mg.The average dose was 4840 mg,2500-7200 mg were the main dose range.More than 5 cycles and (or) more than 2500-7200 mg of carboplatin administration significantly increased the incidence of hypersensitivity reactions in the twelve patients.Beactions were generally occurred at the first 5-10 minutes during intravenous infusion.The average time was 7.6 minutes.Symptoms included mild-to-moderate reactions and severe reactions.Thirteen patients experienced earboplatin hypersensitivity.Two out of 13 cases exhibited severe hypersensitivity reaction at the first time.The first hypersensitivity reactions was mild-to-moderate in 11 cases.When retreated with carboplatin,4 exhibited no more reactions,5 exhibited mild-to-moderate hypersensitiviry reactions,2 exhibited severe reactions.Mildto-moderate reactions were resolved by temporary interruption of carboplatin infusion,and (or) using steroid,while severe hypersensitivity reactions were resolved by more medicines.Conclusions The hypersensitivity reactions in the patients receiving carboplatin are increased after multiple doses of the agent.The possible of retreat with the carboplatin for the mild-to-moderate reactions may be considered.Hypersensitivity reactions should be treated actively.The following chemotherapy should be planed individually.The primary chemotherapy protocol for the patients with severe hypersensitivity reactions should not be reconsidered.

Objective To study the clinical characteristic, the optimal treatments and the prognosis for the recurrence and failure of primary treatment in malignant ovarian germ cell tumors (MOGCT).Methods The clinical data of 17 recurrent and failure of primary treatment in MOGCT cases treated in Pecking Union Medical College Hospital from January 1983 to May 2008 were analyzed retrospectively to evaluate failure of primary treatment and second treatment. Results Only the 4 eases of recurrent and failure of primary treatment of MOGCT were underwent comprehensive surgical staging. After primary surgery in 1 -8 months, 16 cases received the non-standard chemotherapy were found the lesion again. The secondary debulking surgery was done for the 15 cases and also received the standard chemotherapy. Among of them, 8 cases were survival during follow up, 5 cases gave up the treatment and 4 patients were lost following up during the treatment. Conclusions The standard primary treatment is the most important for the MOGCT. Even for the recurrence and failure of primary treatment of MOGCT, the satisfied cytoreduetive surgery plus the standard chemotherapy also show the significant impact on the prognosis.

Objective To compare the effect between intra-arterial and intra-venous neoadjuvant chemotherapy(NACT)in stage Ⅰb2-Ⅱ b cervical carcinoma.Methods A retrospective analysis Was done on 52 cases of intra-venous NACT and 95 eases of intm-arterial NACT for stage Ⅰ b2-Ⅱ b cervical carcinoma treatad in Peking Union Medical College Hospital from 1999.ResulIs The response rate of intraveHous NACT and intra-arterial NACT was 88%(46/52)and 79%(75/95).and the operative rate after NACT Was 81%(42/52)and 72%(68/95)respectively(P>0.05).There were no significant differences in surgery time,blood loss and pest-operative morbidity between these two groups.Pathological parametrial positive rate after NACT in arterial group(6%)Was significantly lower than that of venous group (50%,P>0.05).The venous group had very similar recurrence rates(13%vs 17%)and death rates (9%VS 12%)when compared with the arterial group(P>0.05).Conclusions The intra-arterial and intra-venous NACT for stage Ⅰ b2-Ⅱb cervical carcinoma show similar response rate.operative rate and surgical difficulties.Arterial NACT shows a better effect on parametrial infiltration.