Objective To investigate the etiology,clinical features and prognosis of neonatal gastric perforation.Methods The medical records of 18 patients with neonatal gastric perforation with respect to sex,age,birth-weight,course of disease,clinical presentations,and prognosis were retrospectively analyzed.Results There were 13 boys and 5 girls,8 of whom were full term infants and 10 preterm infants.The most common initial manifestations were poor activity,abdominal distension,and respiratory distress.All neonates were treated by surgical operation,the overall survival rate was 11/18.Prematurity,birth-weight,course of disease,were the statistically significant risk factors (P ＜ 0.05).Conclusions Neonatal gastric perfora tion is mainly caused by congenital gastric wall defect,and associated with high mortality,particularly in premature infants.It is necessary to early diagnosis,surgical operation in time,nurse intensively,and observe the condition closely.

Objective To evaluate the effect of curcumin pretreatment on c-Jun N-terminal kinase (JNK) signaling pathway during one-lung ventilation (OLV)-induced acute lung injury in mice.Methods Ninety SPF male C57BL/6J mice,aged 6-9 weeks,weighing 18-24 g,were randomly divided into 6 groups (n=15 each) using a random number table:two-lung ventilation (TLV) group;OLV group;curcumin 100,150,200 and 250 mg/kg groups (C100,C150,C200 and C250 groups).The corresponding doses of curcumin were administered intraperitoneally at 2 h before one-lung ventilation in C100,C150,C200 and C250 groups.The animals were tracheally intubated and mechanically ventilated in volume-controlled mode.The ventilator settings were adjusted to maintain the end-tidal pressure of carbon dioxide at 35-45 mmHg.In OLV,C100,C150,C200 and C250 groups,unilateral lung was ventilated for 1.5 h followed by 0.5 h of TLV.Bilateral lungs were ventilated for 2.0 h in group TLV.Peak airway pressure and airway pressure were recorded at 1.5 h of OLV and 0.5 h of TLV.At the end of mechanical ventilation,left lungs were removed for microscopic examination of the pathologic changes,and the index of quantitative assessment for alveolar damage (IQA) was recorded.Wet/dry lung weight ratio (W/D ratio) was determined,and the cell apoptosis in lung tissues was detected using TUNEL.The apoptosis index (AI) was calculated.The expression of JNK mRNA was determined using real-time polymerase chain reaction.The expression of JNK and phosphorylated JNK was determined by Western blot.The phosphorylation of JNK was calculated.Results Compared with group TLV,the IQA,W/D ratio,AI,expression of JNK mRNA and phosphorylation of JNK were significantly increased in group OLV (P＜0.05).Compared with group OLV,the IQA,W/D ratio,AI,expression ofJNK mRNA and phosphorylation of JNK were significantly decreased in C150,C200 and C250 groups,the parameters mentioned above were significantly decreased in sequence in C100,C150,C200 and C250 groups (P＜0.05),and no significant change was found in the parameters mentioned above in group C100 (P＞ 0.05).Compared with group OLV,the pathological changes were significantly attenuated in sequence in C150,C200 and C250 groups.Conclusion The mechanism by which curcumin pretreatment reduces cell apoptosis during OLV-induced acute lung injury is related to inhibition of JNK signaling pathway activation in mice.

Objective To improve the diagnostic quality of tumors composed of smooth muscles in alimentary tract.Methods The clinical and X-ray date of 40 cases of smooth musles tumors confirmed by surgery and pathological examination.Results Fifteen cases in the esophagus (including leiomyoma 14,leiomyosarcoma 1)showed the following X-ray appearence:filling defect,unfolding of mucous membrane,widening of the canal and pericanal mass 25 cases in the stomach and intestine(including leiomyoma 6,leiomyoplastoma 7,leiomyosarcoma 12)showed the following X-ray changes depending on the way of tumor,grows,cyst formation caused by tumor necrosis,compression of the stomach or intestine and adjacent organs,rarely seen calcification in tumor.The X-ray changes of small intermascular leiomyoma were short of specific X-ray signs.Conclusion X-ray exeamination is a reliable diagnostic method for smooth muscles tumors in alimentary tract.

AIM: To study the basic mechanism of transcriptional regulation,NKX3.1 gene promoter was cloned and its promoter activities in prostate cancer cell lines and other cancer cell lines were tested.METHODS:(1.04 kb)-promoter fragment of NKX3.1 gene was obtained by PCR and cloned into pGL_3-basic and pEGFP-1 that are promoter-less reporter vectors to examine its promoter activity driving the reporter gene transcription.The promoter activity was determined by dual-luciferase reporter assay and the expression of GFP reporter observed under fluorescence microscope.RESULTS: The sequence of the cloned(1.04 kb) promoter proved to be correct by DNA sequencing.The dual-luciferase reporter assay(M1/M2) showed that the promoter activity in LNCaP cell transfected with pGL_3-(1.04 kb) promoter was about 1.5-fold higher than that of pGL_3-control transfection and 50-fold higher than that of pGL_3-basic transfection.To investigate the 1.04 kb-promoter activity in different tumor cell lines,the constructed pGL_3-(1.04 kb) promoter and pEGFP-1.04 kb promoter were transfected into several cell lines,respectively.The results showed that the activity of(1.04 kb) promoter in LNCaP was highest among the tested cell lines.Multiple consensus sequence elements have been identified within the(1.04 kb) fragment using TRANSFAC database.Further experiments will be done to determine their founctions.CONCLUSION: Cloned 1.04 kb fragment upstream of NKX3.1 gene presented a strong promoter activity and its activity was highest in LNCaP cell among the tested tumor cell lines.

AIM: To study the basic mechanism of transcriptional regulation, NKX3.1 gene promoter was cloned and its promoter activities in prostate cancer cell lines and other cancer cell lines were tested. METHODS: 1.04 kb - promoter fragment of NKX3. 1 gene was obtained by PCR and cloned into pGL3 - basic and pEGFP - 1 that are promoter - less reporter vectors to examine its promoter activity driving the reporter gene transcription. The promoter activity was determined by dual -luciferase reporter assay and the expression of GFP reporter observed under fluorescence micro scope. RESULTS: The sequence of the cloned 1.04 kb promoter proved to be correct by DNA sequencing. The dual - lu ciferase reporter assay (M1/M2) showed that the promoter activity in LNCaP cell transfected with pGL3 - 1.04 kb promoter was about 1.5 - fold higher than that of pGL3 - control transfection and 50 - fold higher than that of pGL3 - basic transfec tion. To investigate the 1.04 kb - promoter activity in different tumor cell lines, the constructed pGL3 - 1.04 kb promoter and pEGFP - 1.04 kb promoter were transfected into several cell lines, respectively. The results showed that the activity of 1.04 kb promoterin LNCaP was highest among the tested cell lines. Multiple consensus sequence elements have been iden tified within the 1.04 kb fragment using TRANSFAC database. Further experiments will be done to determine their founc tions. CONCLUSION: Cloned 1.04 kb fragment upstream of NKX3.1 gene presented a strong promoter activity and its activity was highest in LNCaP cell among the tested tumor cell lines.

AIM: To observe the effect of exogenous androgen responsive element decoy on the promoter of prostate specific antigen (PSA) and the growth of LNCaP cells for searching the possibility of gene therapy for prostate cancer. METHODS: Firstly, pGL3 - PSA luciferase expression vector containing 640bp - promoter fragment of PSA gene was constructed. Then, a 23 -mer phosphorothioated ARE decoy based on the deduced ARE sequence at the promoter region of PSA gene was synthesized. pGL3 - PSA and ARE decoy DNA were cotransfected into PC3 - M cell by lipofectamineTM 2000. Through detecting the activity of luciferase, the effect of ARE decoy on the promoter of PSA was studied. Then the ARE decoy DNA was transfected into LNCaP cells. The effect of decoy DNA on the proliferation of LNCaP cells was examined by using MTT assay. The effects of apoptosis were detected by phase contrast microscopy, DNA agrose gel electrophoresis and flow cytometry. Meanwhile, the nuclear extract was prepared from LNCaP cells and DNA - protein interactions were examined by electrophoretic mobility shift assay (EMSA). RESULTS: The reporter assay showed that the aetivity of luciferase was significantly reduced in the ARE decoy - transfected cells, bnt not in the cells transfected with the control decoy. EMSA demonstrated specific binding of the ARE decoy to androgen receptor. The growth of LNCaP was remarkably inhibited and apoptotic morphological changes as well as DNA fragmentation were observed in the ARE decoy- transfected cells. The rate of apoptosis was 22.4% detected by FCM. CONCLUSION: The ARE decoy is capable of inhibiting the promoter of PSA gene and inducing the apoptosis in prostate cancer cells. It may become a potential therapeutic tool for prostate cancers.

AIM: To observe the effect of E2F decoy DNA on proliferation and apoptosis of androgen-independent prostate cancer cell line PC-3M.METHODS: E2F decoy DNA,ARE decoy DNA and control decoy DNA were transfected into PC-3M cells with lipofectamine,respectively.Their effects on cell proliferation were detected by MTT assay.The changes of cell morphology were observed by inverted phase contrast microscope.The cell apoptotic rate was determined by flow cytometry(FCM) analysis and chromosome DNA ladder was detected by DNA gel electrophoresis.The expression of c-Myc mRNA and cyclin D1 mRNA was detected by RT-PCR.The protein levels of c-Myc and cyclin D1 were detected by Western blotting.RESULTS: The growth of PC-3M cells was inhibited after transfection.The transfected PC-3M cells displayed typical apoptotic morphological changes.The apoptotic rate was 26.35% and DNA ladder was observed after transfection.The expression of c-Myc and cyclin D1 were inhibited.CONCLUSION: These results indicate that E2F decoy DNA induces apoptosis of androgen-independent prostate cancer cell lines PC-3M and inhibits cell proliferation via inhibiting expression of c-Myc and cyclin D1.

Objective To evaluate the effect of dezocine on the c-fos expression in neurons in the midbrain periaqueductal gray in a rat model of incisional pain.Methods Thirty-six pathogen-free healthy adult male Wistar rats,weighing 250-300 g,were divided into 3 groups (n =12 each) using a random number table:control group (group C),incisional pain group (group I) and dezocine group (group D).A 1 cm longitudinal incision was made through skin,fascia and muscle of the plantar aspect of the right hind paw in sevoflurane-anesthetized rats.In group C,the rats were only anesthetized and underwent no operation.In group I,0.9％ sodium chloride solution 2 ml was injected via the caudal vein at 15 min before the model was established.In group D,dezocine 1 mg/kg (diluted to 2 ml in 0.9％ sodium chloride solution) was injected via the caudal vein at 15 min before the model was established.At 24 h before operation (T0) and 2,6 and 24 h after operation (T1-3),the mechanical paw withdrawal threshold (MWT) and cumulative pain score were measured.After measurement of the pain threshold at T3,the whole brain was removed for determination of the c-fos expression in neurons in the midbrain periaqueductal gray by immunohistochemistry.Results Compared with group C,the MWT was significantly decreased,cumulative pain scores were increased,and the expression of c-fos in neurons in the midbrain periaqueductal gray was upregulated at T1-3 in I and D groups (P＜0.05).Compared with group I,the MWT was significantly increased,the cumulative pain score was decreased,and the expression of c-fos protein in neurons in the midbrain periaqueductal gray was down-regulated at T1.3 in group D (P＜0.05).Conclusion Dezocine mitigates incisional pain through inhibiting the expression of c-fos in neurons in the midbrain periaqueductal gray of rats.

AIM: To investigate the effects of protein C activator (PCA) from Agkistrondon acutus venom ( AAV) on the tension of thoracic aorta rings isolated from the rats with sepsis.METHODS:The model of sepsis was es-tablished by intraperitoneal injection of lipopolysaccharide ( LPS) .SD rats were randomly divided to 6 groups ( n=6 ):sham group, LPS group, PCA intervention group (LPS+PCA, PCA at doses of 0.1 mg/kg, 0.3 mg/kg and 0.6 mg/kg) and LPS+polymyxin B (at dose of 0.2 mg/kg) group.Using perfusion experiment in vitro, the tension of the aortic rings was measured by biological signal analytical system.RESULTS:The values of MABP, HR, LVDP and ±dp/dtmax were significantly lower in LPS group than those in sham group and LPS+PCA groups.Compared with sham group, the relaxa-tion response to acetylcholine ( ACh) and the contractile response of aorta rings induced by phenylephrine ( Phe) were sig-nificantly decreased in LPS group, which were increased significantly in PCA intervention group ( especially at dose of 0.6 mg/kg) compared with LPS group.The dose-response curve of aorta contraction with denuded endothelium induced by Phe shifted down significantly in LPS group compared with sham group, and no significant difference between LPS group and PCA intervention group was observed.Also no statistical difference was found in non-endothelium dependent relaxation of aortic rings induced by sodium nitroprusside among the groups.Pretreatment of N-nitro-L-arginine methl ester and methyl-ene blue increased the contraction amplitude of aortic rings induced by Phe.CONCLUSION:PCA from AAV effectively reverses the hypoergia of the vessels in rats with sepsis through protecting vascular endothelium, the mechanism of which may be mediated by inhibiting NO-GC-cGMP signal transduction pathway.

Objective To investigate the effect of hyperbaric oxygen therapy on serum procalcitonin (PCT), white blood cells(WBC) and high-sensitivity C-reactive protein (hs-CRP) levels in children with necrotising fasciitis (NF) and its efficacy, and explore the diagnosis value of above indicators.Methods From March 2011 to June 2014,50 cases children with acute necrotic fasciitis treated in Children's Hospital of Hebei Province as study group,which were randomly divided into hyperbaric oxygen group (n =25) and routine group (n =25) .The routine group received the routine therapy of incision and drainage to clear the lesion, hyperbaric oxygen group received hyperbaric oxygen therapy on the basis of routine group,while 50 healthy children were selected as control group.The serum PCT, WBC, hs-CRP levels, efficacy, complications, death and hospitalization time were observed and compared.Results The serum PCT, WBC and hs-CRP levels pre-treatment in study group were higher than those in control group(P<0.05).The area under the ROC curve of PCT and hs-CRP was 1.000,respectively, and WBC was 0.804, there were significant difference between PCT and WBC (Z=5.250,P=0.000), between hs-CRP and WBC (Z=5.037,P=0.000).After treatment, the wounds of 23 case patients (92.00%) were cured in hyperbaric oxygen group, and 21 cases in routine group (84.00%) , there were no significant difference in cure rate between two groups.There were six cases(24.00%) of complications and one case (4.00%) of death in hyperbaric oxygen group,while nine cases (36.00%) of complications and two cases (8.00%) of death, there were no significant difference in complications rate and death rate between two groups.The hospitalization time in hyperbaric oxygen group was (39.17 ±6.73) d, which was significantly lower than (52.13 ±4.28) d in routine group(P<0.05).Conclusion PCT and hs-CRP have certain value in diagnosis of children with acute necrotizing fasciitis; incision and drainage combined with hyperbaric oxygen therapy has a better clinical effect in the treatment of children with acute necrotizing fasciitis.