Objective To observed the expression of matrix metalloproteinase-9 in the early onset of severe acute pancreatitis associated with acute lung injury in rats and investigate its effection in lung injury.Methods Thirty-two healthy adult male SD rats were randomly divided into two groups:Control group (n =8),Severe acute pancreatitis group(n =24).Severe acute pancreatitis model was induced by retrograde inject the 4％ sodium taurocholate sodium taurocholate into the biliopancreatic duct of rats.The severe acute pancreatitis group was detected the rate of lung water content、arterial blood gas.myeloperoxidase,matrix metalloproteinase-9,histopathology of the pancreas and lung injury score under the light microscope at 3 hours,6 hours and 12 hours.The matrix metalloproteinase-9 expression was detected by immunohistochemical and the results of immunohistochemical were analysed by the Image-Pro Plus image analysis system.Control group was detected the relevant indicators at 12 hours.Results Successfully modeling,the expression of matrix metalloproteinase-9 gradually increased beginning at 3 hours,at twelve hours up to the highest value(P ＜ 0.05).The degree of lung injury,lung water content,myeloperoxidase activity,PaCO2 gradually increased(P ＜ 0.05),PaO2 decreased significantly P ＜ 0.05).Conclusions The high expression of matrix metalloproteinase-9 is important to the pathogenesis of severe acute pancreatitis associated with acute lung injury.

Objective To investigate the relationship of point mutations in ERG11,TAC1,MRR1 and UPC2 genes to fluconazole resistance in Candida albicans.Methods This study included 11 fluconazole-resistant Candida albicans strains.DNA was extracted from these Candida isolates by urea pyrolysis method.PCR was carried out to amplify the full-length EGR11 gene,as well as the corresponding fragments of TAC1,MRR1 and UPC2 genes followed by DNA sequencing.Results Eight mutations were detected in the ERG11 gene,including D116E,Y132H,Y132F,K143Q,T229A,E266D,G448E and G464S.Among them,K143Q was a novel mutation and had not been reported.A G648D mutation was detected in the UpC2 gene.No mutations were detected in the TAC1 or MRR1 gene.Conclusions There is a definite association between fluconazole resistance and ERG11 mutations,while further research is warranted to clarify the relationship between the mutations of TAC1,MRR1,UPC2 genes and resistance to fluconazole.

Objective To localize the tibial attachment of the posterior cruciate ligament (PCL) on the magnetic resonance imaging (MRI) and provide parameters for clinical PCL reconstruction.Methods We retrospectively analyzed 524 patients with intact tibial PCL attachment who had undergone knee MRI from January 2010 to January 2016.They were 286 men and 238 women with an average age of 35 years (from 20 to 50 years).The size and positions of the tibial PCL attachment were measured on the sagittal and coronal MRI slices.The differences were analyzed between different genders.Results On the sagittal slices,the mean distance from the central tibial PCL attachment to the posterior edge of the tibial plateau was 17.9 ± 3.0 mm and the mean anteroposterior diameter of the tibial PCL attachment was 9.7 ± 2.4 mm,with those for males significantly larger than for females (P ＜ 0.05).The above mean values when expressed as a percentage of the posterior tibial slop were 79.9％ ±4.5％ and 43.7％ ± 9.6％,respectively,showing no significant differences between males and females (P ＞ 0.05).On the coronal slices,the distances from the central tibial PCL attachment to the medial and lateral edges of the tibial plateau were 33.5 ± 3.1 mm and 37.4 ±4.1 mm,respectively,and the mediolateral diameter of the tibial PCL attachment was 12.0 ± 1.6 mm,with those for males significantly larger than for females (P ＜ 0.05).The above mean values when expressed as a percentage of the mediolateral diameter of the tibial PCL attachment were 47.4％ ± 3.2％,52.7％ ±3.1％ and 16.9％ ± 1.7％,respectively,showing no significant differences between males and females (P ＞ 0.05).Conclusions On knee MRI images,the distance from the central tibial PCL attachment to the posterior edge of the tibial plateau is about 17.9 mm,the anteroposterior diameter of the tibial PCL attachment around 9.7 mm,and the mediolateral diameter of the tibial PCL attachment roughly 12.0 mm.These measurements for males are larger than for females.

Objective To investigate the inhibitory effect of IN5 from chlamydiaphage phiCPG1 capsid protein Vp1 on Chlamydia trachomatis (Ct).Methods PCR was used to amplify IN5 gene from Vp1 DNA of phiCPG1, then the recombinant plasmid pET28a/IN5 was constructed.After transformation, the fusion protein IN5 was induced,identified and purified.Ct was incubated with the purified IN5 protein or Vp1 protein.After 48 h of incubation, the inclusion bodies were counted with iodine staining and indirect immunofluorescence.One-way ANOVA was used to compare the difference of inclusion bodies among groups.If the difference among the groups was statistically significant, the Bonferroni method was used to compare any two mean values.Finally, the inhibitory rate of IN5 protein and Vp1 protein to Ct was calculated.Results IN5 protein from chlamydiaphage phiCPG1 capsid protein Vp1 was successfully obtained.At the same concentration of 53μg/mL,the inhibitory rates of Ct growth in IN5 and in Vp1 groups were 52.42% and 78.04%, respectively.Conclusion IN5 protein has inhibitory effect on the growth of Ct,but the inhibitory rate is lower than that of Vp1, which provides a preliminary clue for searching the dominant region of Vp1 protein inhibiting the growth of Ct.

Objective To clone and express the polymorphic membrane protein I(PmpI)gene of Chlamydia trachomatis(Ct), and to assess the immunogenicity and biological characteristics of PmpI. Methods A bioinformatic software was used to analyze the sequence of the PmpI gene of Ct, and to predict B cell epitopes in PmpI. With Ct serovar D DNA as the template, PCR was performed to amplify the N?terminal region(from position 90 to 1464)of the PmpI gene, which was cloned into a prokaryotic expression vector pET28a to express the recombinant protein PmpI. A Ni?ion affinity chromatography column was used to purify the recombinant protein, which was used to immunize New Zealand rabbits for preparation of polyclonal antibodies. Western blot analysis was conducted to evaluate the immunogenicity of this protein. Results A comprehensive analysis was carried out on the secondary structure, flexible regions, hydrophilicity plot, antigenic index and surface probability plot of the protein, which suggested that PmpI had 8 dominant B?cell epitopes. The product of PCR targeting the PmpI gene of Ct serovar D showed a total length of 1 375 bp. The recombinant prokaryotic expression vector pET28a?PmpI was successfully constructed. A recombi?nant protein with a relative molecular mass of approximately 50 000 was successfully expressed after isopropylβ?d?1?thiogalactopyranoside (IPTG) induction, and purified by affinity chromatography. Polyclonal antibodies against the recombinant protein were successfully prepared. Conclusion The N?PmpI protein of Ct serovar D is cloned and expressed successfully, laying a foundation for further studies on its biological functions.

AIM: To investigate the role of NF-?B/I?B signal pathway in the regulation of (cyclooxygenase-2) (COX-2) expression in human mesangial cells (HMC). METHODS: The PGE_2 concentration in supernatants of HMC was measured by radioimmunoassay. COX-2 mRNA and protein expression were determined by RT-PCR and Western blot. Electrophoretic mobility shift assay (EMSA) and Western blot were used to detect the activity of NF-?B and degradation of I?B. RESULTS: IL-1? significantly upregulated COX-2 expression and PGE_2 production in HMC. Significant up-regulation of NF-?B activation, nuclear translocation of p65 subunit, and degradation of I?B ? and I?B ? were observed in IL-1?-induced HMC. CONCLUSION: Expression of COX-2 in IL-1?-induced HMC is mediated by NF-?B/I?B signal pathway. [

Objective To compare the biomechanical stability of 4 internal fixations in treatment of acetabular fracture of the lower anterior column through finite element analysis.Methods One normal adult male pelvis was subjected to 0.7mm thin-section CT scanning and 379 CT pictures were obtained.Finite element modeling software was used to establish internal fixation models for acetabular fracture of the lower anterior column,including lag screws (A),anterior column reconstruction plate (B),subcutaneous plate not crossing the pubic symphysis (C) and subcutaneous plate crossing the pubic symphysis (D).Finite element analysis was carried out to compare the biomechanical differences among the 4 internal fixation models which were subjected to the same loading conditions at both standing and sitting positions.Results At standing and sitting positions,the maximum displacement and the mean node displacement of fracture lines were the greatest in group A (0.558 mm and 0.462 ±0.092 mm at standing;0.634 mm and 0.473 ±0.108 mm at sitting),the smallest in group D (0.512 mm and 0.425 ±0.083 mm at standing;0.031 mm and 0.025 ± 0.004 mm at sitting),and in between in group B (0.513 mm and 0.432 ±0.085 mm at standing;0.630 mm and 0.466 ± 0.109 mm at sitting) and in group C (0.514 mm and 0.433 ± 0.085 mm at standing;0.627 mm and 0.464 ± 0.107 mm at sitting).At both standing and sitting positions,the maximum stress at the fracture line was the greatest in group D (10.519 MPa and 24.879 MPa),the smallest in group A (3.254 MPa and 8.954 MPa),and in between in group B (4.873 MPa and 9.431 MPa) and in group C (4.384 MPa and 10.128 MPa).Conclusions In treatment of acetabular fracture of the lower anterior column,subcutaneous plate crossing the pubic symphysis may result in the greatest biomechanical stability,lag screws the smallest biomechanical stability,and anterior column reconstruction plate and subcutaneous plate not crossing the pubic symphysis the moderate biomechanical stability.

Objective To evaluate inhibitory effects of the Chlamydiaphage phiCPG1 capsid protein Vp1 on Chlamydia psittaci strain guinea pig inclusion conjunctivitis (GPIC) and Chlamydia trachomatis (Ct) serovar E,and to provide new ideas for the treatment of Ct infection.Methods The Chlamydiaphage phiCPG1 capsid protein Vp1 was expressed in Escherichia coli BL21 transfected with the recombinant plasmid Vp1-pET30a (+),identified by Western blot analysis and purified by using dialysis bags.Bicinchonininc acid (BCA) assay was performed to determine the concentration of Vp1 protein.GPIC and Ct serovar E strains were both classified into 4 groups to be firstly incubated with Vp1 protein (Vp1 group),Tris-glycine solution (Tris group),S protein (S group) or Dulbecco's Modified Eagle Medium (DMEM,DMEM group) at room temperature for 3 hours,then were used to infect Hela cells followed by 72-hour (GPIC) or 48-hour (Ct serovar E) culture with the presence of Vp 1 protein (Vp 1 group),Tris-glycine solution (Tris group),S protein (S group) or DMEM (DMEM group).Subsequently,immunofluorescence staining was conducted to observe and count chlamydial inclusions.Results The number of GPIC inclusions was significantly different between the 4 groups after 72-hour culture (F=476.632,P＜ 0.05),and lower in the Vp1 group (5.0 ± 1.5) than in the Tris group (24 ± 1.2,P＜ 0.05),S group (25 ± 1.7,P＜ 0.05) and DMEM group (25 ± 1.5,P＜ 0.05),but insignificantly different between the latter 3 groups (P ＞ 0.05).Compared with the DMEM group,the Vp1 group showed a significant decrease of 80.2％ ± 3.99％ and 77.2％ ± 1.79％ in the number of GPIC and Ct serovar E inclusions respectively,with no significant difference in the inhibitory effect of Vp1 on GPIC versus Ct serovar E (t =2.057,P ＞ 0.05).Conclusion The phiCPG1 capsid protein Vp1 can obviously inhibit GPIC and Ct serovar E infections to a similar degree.

Objective:To explore independent predictors of postoperative delirium in elderly patients by PRe-Operative Prediction of postoperative DElirium by appropriate SCreening (PROPDESC) and Mayo Delirium Prediction (MDP) , and analyze the predictive power of the two models.Methods:This study was a prospective Cohort study. Using the convenient sampling method, a total of 636 elderly surgical patients admitted to the Orthopedics, Gastroenterology, Cardiothoracic and Oncology Departments of Shantou Central Hospital from May to August 2022 were selected as the research objects. PROPDESC and MDP were used to predict postoperative delirium in elderly patients. The area under the receiver operating characteristic ( AUC) and diagnostic characteristics of the two predictive models were compared, and the single factor analysis and Logistic regression analysis were performed on 19 predictive factors of the two models to determine the independent influencing factors of postoperative delirium. Results:The AUC for external verification of the PROPDESC and MDP were 0.87 (95% CI: 0.84-0.90) and 0.89 (95% CI: 0.86-0.92) , respectively. The sensitivity of 71.79% and 80.34%, and specificity of 85.16% and 81.12%, respectively. Logistic regression analysis showed that emergency admission, age, sentence repetition and sequence subtraction in Montreal Cognitive Assessment (MoCA) were independent influencing factors for postoperative delirium ( P<0.05) . Conclusions:The predictive ability of PROPDESC and MDP models to predict postoperative delirium in elderly patients is satisfactory. On this basis, delirium risk assessment tools suitable for different surgical elderly populations in China can be constructed.

Objective To construct active phages against Chlamydia trachomatis,and to evaluate its effect on Chlamydia trachomatis.Methods The M13 phage was recombined with the IN5 sequences encoding the capsid protein VP1 of chlamydiophage phiCPG1,and then the recombinant M13-IN5 phage was obtained.PCR amplification,enzyme digestion and sequencing were performed to verify whether the target fragment was inserted into the phage successfully.The viability of the phage was evaluated by plaque formation assay.Cell counting kit-8 (CCK8) assay was conducted to evaluate the effect of M13 phage and recombinant M13-IN5 phage at the titer of 1011 plaque-forming units (PFU)/ml on the proliferation of Hela cells,and Hela cells uninfected with chlamydia served as the blank control group.Western blot analysis was performed to determine the expression of the IN5 loop protein in the recombinant M13-IN5 phage,M13 phage and Escherichia coli ER2738 at exponential growth phase.Cultured standard Chlamydia trachomatis serovar E strain was treated with M13 phage and recombinant M13-IN5 phage at the titer of 1011 PFU/ml separately,and chlamydia control group without the treatment with phages was set up.After 36-hour infection,confocal microscopy was performed to detect the location of the M13 phage and the recombinant M13-IN5 phage.Moreover,iodine staining was conducted to count inclusion bodies at 36,48,60 and 72 hours separately after infection.Statistical analysis was carried out by a two-sample t-test for comparisons between two groups,one-way analysis of variance (ANOVA) for intergroup comparison,and Bonferroni test for multiple comparisons.Results The bioactive recombinant M13 phage containing the IN5 loop gene was constructed successfully,and Western blot analysis confirmed that the recombinant phage expressed IN5 loop/p Ⅲ fusion protein with a high titer of 3.05 × 1011 PFU/ml.As CCK8 assay showed,there was no significant difference in proliferation of Hela cells among the blank control group,M 13 phage group and recombinant M13-IN5 phage group (A450 values:3.63 ± 0.01,3.55 ± 0.02,3.70 ± 0.01,respectively,F =12.0,P ＞ 0.05).Confocal microscopy showed overlap between the phage fluorescence and chlamydial inclusion body fluorescence.The M13-IN5 phage group and M13 phage group both showed significantly decreased number of inclusion bodies compared with the control group (both P ＜ 0.05) at 36 and 72 hours after chlamydial infection,and the number of inclusion bodies was significantly lower in the M 13-IN5 phage group than in the M13 phage group (P ＞ 0.05).After 48,and 60 hours of chlamydial infection,the number of inclusion bodies did not differ among the M13 phage group,M13-IN5 phage group and control group (both P ＞ 0.05).Conclusions The recombinant M13-IN5 phage was bioactive and could successfully express the IN5 loop protein.In the in vitro experiments,the recombinant phage could enter into chlamydia inclusion bodies,and markedly inhibited the infection of Chlamydia trachomatis.