OBJECTIVE To investigate the distribution of multiresistant genes produced by Pseudomonas aeruginosa(PAE) isolated from different areas.METHODS Antimicrobial sensitivities of 104 strains of PAE to 13 kinds of antibiotics were determined by K-B method,of which 16 kinds of drug resistant genes were tested by polymerase chain reaction(PCR) and analyzed by clustering method of multi-molecular markers.RESULTS The resistant detective rates to imipenam in three areas were respectively 51.3%,80%,and 100% and that to the third generation cephalosporin were moving from 48.2% to 100%.The detective rate of oprD2 absent was 90-100%.There were similar phenomena of prevalence between some strains in these areas.oprD2 Absent was the main mechanism of prevailing strains in Shaoxing,however,clone strains in Huzhou were caused by TEM,oprD2 absent,aac(3)-Ⅱ,ant(3″)-Ⅰ and qacE△1.CONCLUSIONS There is a multiresistance to antibiotics in PAE isolated from different areas and the phenomena of prevalence are occurred in these areas,but the mechanisms of drug resistant are different.

Objective To explore the possibility of guiding clinical fast diagnosis and rational use of drugs by rapid detection of plasma digoxin concentration(PDC) with i4000.Methods The plasma samples of 132 patients were collected and digoxin concentration was detected by i4000,and the relationship between PDC and clinical effect was analyzed.Then,the regularities of distribution of digoxin concentration also analyzed according to the age and PDC.The patients whose PDC beyond the effective concentration 0.8 ～ 2.0 μg/L,were treated with adjusted dose respectively according to the circumstance and continuous monitoring of PDC.Results In 132 cases,PDC of 106 patients within the therapeutic dose,accounted for 80.30％,and the total effective rate was 86.36％ after treatment.The effective rates in ＜0.8μg/L group,0.8 ～2.0μg/L group and ＞2.0μg/L group were 10.91％,95.28％and 75.00％,respectively.After dosage adjusted for 1 1 cases with PDC ＜ 0.8μg/L and 4 cases ＞ 2.0μg/L,the PDC returned to the effective concentration.The PDC in over 60 years old group was higher than that in 50 ～ 60 years old group.Poisoning symptoms occurred in 7 cases,and symptoms disappeared through adjustment dosages.Conclusion The PDC detection by Abbott i4000 is rapid and easy to operate,and the result is accurate and reliable.

OBJECTIVETo obtain transgenic Pinellia ternata plants resistant to fungus by transfer Chitinase and beta-1,3-Glucanase gene from Trichoderma harzianum.

METHODUsing hygromycin phosphotransferase as the selection marker, the Chitinase gene (ech42), beta-1,3-Glucanase gene (gluc78) and both gene pCAMBIA(ech42 + gluc78) driven by CaMV35S promoter were transferred into P. ternata callus via Agrobacterium-mediated transformation.

RESULTPCR results confirmed that the regenerants were identified to be transgenic lines and the RT-PCR results confirmed that foreign genes construction were transfer to mRNA. Two foreign genes were inherited stably to T5 generation according to PCR results of the lines.

CONCLUSIONThe results showed that chitinase gene ech42 and beta-1, 3-glucanase gene gluc78 respectively or together introducing and co-integrating into P. ternata

Agrobacterium tumefaciens ; genetics ; metabolism ; Chitinases ; genetics ; metabolism ; Fungal Proteins ; genetics ; metabolism ; Gene Expression Regulation, Plant ; Gene Transfer Techniques ; Genetic Vectors ; genetics ; metabolism ; Glucan 1,3-beta-Glucosidase ; genetics ; metabolism ; Pinellia ; genetics ; metabolism ; Transformation, Genetic ; Trichoderma ; enzymology

Objective:To observe the efficacy the application of endoscope assisted temporoparietal fascia flap harvest for the second-stage operation in auricular reconstruction of Nagata’s technique for microtia.Method:In this retrospective study, the clinical data were collected from the patients who received microtia reconstruction with autologous rib cartilage at the Department of Burns and Plastic Surgery, General Hospital of Northern Theater Command from January 2015 to January 2022. According to the surgical procedure, patients were divided into endoscopic group and open surgery group. In endoscopic group, endoscope-assisted temporoparietal fascia harvest were performed for the second-stage operation in auricular reconstruction of Nagata’s technique. In open surgery group, temporoparietal fascia flaps were harvested in open surgery for the second-stage operation in auricular reconstruction of Nagata’s technique. Regular follow-up was conducted to observe the survival of the fascia flaps, complications, patient satisfaction, and surgical scars. The patient satisfaction questionnaire for auricular reconstruction was used to assess patient satisfaction, and the patient and observer scar assessment scale (POSAS) was used to evaluate scar formation in the surgical area. Data analysis was performed using SPSS 26.0 statistical software. The measurement data were expressed by Mean ± SD, and the counting data were expressed as cases (%). The T-test was used to compare the age difference, length of hospital stay, intraoperative blood loss, scar length, patient satisfaction, and POSAS scores between the two groups. Chi-square test was used to compare the gender composition and incidence of complications between the two groups. P<0.05 was considered statistically significant. Results:A total of 51 patients were included, with 26 in the endoscopic group (14 men and 12 women) and 25 in the open surgery group (12 men and 13 women). The age of the patients in the endoscopic group was (9.8±2.9) years (ranging from 7 to 17 years), while in the open surgery group was (10.3±3.8) years (ranging from 7 to 17 years). The postoperative follow-up period was (15.4±3.4) months (1 to 2 years), and all fascia flaps survived without any severe complications. There were no statistically significant differences between the two groups in terms of age difference, length of hospital stay, intraoperative blood loss, postoperative satisfaction, sex composition ratio, and postoperative complications ( P>0.05). The scar quality in the endoscopy group was superior to that in the open surgery group, and POSAS scores of endoscopic group were lower than those in the open surgery group, and the difference was statistically significant ( P<0.05). Conclusion:Endoscope assisted temporoparietal fascia flap harvest for the second-stage operation in auricular reconstruction of Nagata’s technique for microtia can minimize scarring, improve the postoperative appearance and is not statistically associated with the appearance of reconstructed auricles or complications.

Objective:To observe the efficacy the application of endoscope assisted temporoparietal fascia flap harvest for the second-stage operation in auricular reconstruction of Nagata’s technique for microtia.Method:In this retrospective study, the clinical data were collected from the patients who received microtia reconstruction with autologous rib cartilage at the Department of Burns and Plastic Surgery, General Hospital of Northern Theater Command from January 2015 to January 2022. According to the surgical procedure, patients were divided into endoscopic group and open surgery group. In endoscopic group, endoscope-assisted temporoparietal fascia harvest were performed for the second-stage operation in auricular reconstruction of Nagata’s technique. In open surgery group, temporoparietal fascia flaps were harvested in open surgery for the second-stage operation in auricular reconstruction of Nagata’s technique. Regular follow-up was conducted to observe the survival of the fascia flaps, complications, patient satisfaction, and surgical scars. The patient satisfaction questionnaire for auricular reconstruction was used to assess patient satisfaction, and the patient and observer scar assessment scale (POSAS) was used to evaluate scar formation in the surgical area. Data analysis was performed using SPSS 26.0 statistical software. The measurement data were expressed by Mean ± SD, and the counting data were expressed as cases (%). The T-test was used to compare the age difference, length of hospital stay, intraoperative blood loss, scar length, patient satisfaction, and POSAS scores between the two groups. Chi-square test was used to compare the gender composition and incidence of complications between the two groups. P<0.05 was considered statistically significant. Results:A total of 51 patients were included, with 26 in the endoscopic group (14 men and 12 women) and 25 in the open surgery group (12 men and 13 women). The age of the patients in the endoscopic group was (9.8±2.9) years (ranging from 7 to 17 years), while in the open surgery group was (10.3±3.8) years (ranging from 7 to 17 years). The postoperative follow-up period was (15.4±3.4) months (1 to 2 years), and all fascia flaps survived without any severe complications. There were no statistically significant differences between the two groups in terms of age difference, length of hospital stay, intraoperative blood loss, postoperative satisfaction, sex composition ratio, and postoperative complications ( P>0.05). The scar quality in the endoscopy group was superior to that in the open surgery group, and POSAS scores of endoscopic group were lower than those in the open surgery group, and the difference was statistically significant ( P<0.05). Conclusion:Endoscope assisted temporoparietal fascia flap harvest for the second-stage operation in auricular reconstruction of Nagata’s technique for microtia can minimize scarring, improve the postoperative appearance and is not statistically associated with the appearance of reconstructed auricles or complications.

OBJECTIVETo explore the molecular pathogenesis for a pedigree affected with coagulation factor Ⅴ (FⅤ) deficiency.

METHODSProthrombin time (PT), activated partial thromboplastin time (APTT), fibrinogen (FIB), coagulation factor Ⅱ activity (FⅡ: C), FⅤ activity (FⅤ: C), coagulation factor Ⅶ activity (FⅦ: C), and coagulation factor Ⅹ activity (FⅩ: C) were determined with a STAGO automatic coagulometer. FⅤ antigen (FⅤ: Ag) was detected with enzyme linked immunosorbent assay (ELISA). All exons and their flanking regions, and 5' and 3' untranslated regions of the F5 gene were analyzed by direct sequencing. Suspected mutation was verified by reverse sequencing as well as testing of family members. ClustalX software was used to analyze the conservative property of the mutation sites. PROVEAN and MutationTaster online software was used to predict the effect of the mutation on the protein function. Swiss-pdbViewer was used to analyze the protein model and interaction of amino acids.

RESULTSThe PT and APTT of the proband were slightly prolonged to 15.2 s and 41.8 s, respectively. And the FⅤ: C and FⅤ: Ag measured 55% and 62%, respectively. The FⅤ: C and FⅤ: Ag of his father and son were decreased to various extent (60%, 65% and 31%, 40%, respectively). A c.911G>A heterozygous mutation (Gly276Glu) was detected in exon 6 of the proband, for which her father and son were heterozygotes. The same mutation was not found in her mother, brother and husband. Conservation analysis showed that the Gly276 is highly conserved across various species. By bioinformatic analysis, the PROVEAN (scored -6.214) indicated Gly276Glu was harmful, and MutationTaster (scored 0.976) suggested that it is pathogenic. Model analysis suggested there are two hydrogen bonds between Gly276 and Ile298 in the wild type protein. When Gly276 was replaced by Glu276, the original hydrogen bond did not change, but the side chain of Glu was extended, which added steric hindrance with the surrounding amino acids, which resulted in decreased protein stability.

CONCLUSIONThe heterozygous c.911G>A (Gly276Glu) mutation of the F5 gene probably underlies the decreased level of FⅤin the proband.

Adult ; Computational Biology ; Factor V ; chemistry ; genetics ; Factor V Deficiency ; genetics ; Female ; Humans ; Male ; Middle Aged ; Mutation ; Pedigree ; Phenotype

Objective：To investigate the effect of long-chain non-coding RNATTTY10 (lncRNATTTY10) on the migration and invasion of cervical cancer cells, and to explore its regulatory effect on miR-490-3p and HMGB1 (high mobility group box 1) signaling pathways. Methods: Fourteen paris of cervical cancer tissues and corresponding paracancerous tissues resected at the Department of Obstetrics and Gynecology,Affiliated Wuhan Central Hospital of Tongji Medical College fromAugust 2013 to December 2014 were collected for this study. The expression of TTTY10 in cervical cancer tissue and different cervical cancer cell lines were detected by qPCR. Plasmids encoding TTTY10-siRNA or empty plasmids were transfected into cervical cancer CasKicells, and the transfection efficiency was detected by qPCR. Transwell migration assay and Transwell invasion assay were used to detect the migration and invasion abilities of cervical cancer cells after TTTY10 silencing. qPCR was used to detect the expression of miR-490-3p and HMGB1 mRNA after TTTY10 silencing. Dual luciferase reporter assay validated the interaction between miR-490-3p and HMGB1. Western blotting was used to detect the expression of HMGB1 signaling pathway related proteins after TTTY10 silencing. Results: The expression of TTTY10 in cervical cancer tissues was significantly higher than that in paracancerous tissues (P<0.01), the expression of TTTY10 in cervical cancer cell lines was significantly higher than that in cervical epithelial cells (P<0.01). TTTY10-siRNAplasmids could efficiently transfectCasKicells to knockdown TTTY10 expression (P<0.01). Silencing of TTTY10 inhibited the migration and invasion of cervical cancer CasKi cells (P<0.05), promoted the expression of miR-490-3p (P<0.01) and inhibited the expression of HMGB1 mRNAin cervical cancer (P<0.05 or P<0.01). miR-490-3p could specifically bind to the 3'-UTR of HMGB1 mRNA(P<0.01). HMGB1 signaling pathway related proteins were down-regulated after TTTY10 silencing. Conclusion: TTTY10 can target regulate the expression of miR-490-3p and affect the migration and invasion ability of cervical cancer CasKi cells through the HMGB1 signaling pathway; TTTY10 can be used as a diagnostic marker and potential treatment target of cervical cancer.