Objective:To analyze the multiple locus variable number tandem repeat analysis (MLVA) genotype polymorphism of Bacillus (B.) anthracis and establish a MLVA genotype database of B. anthracis in China. Methods:B. anthracis strains isolated from different sources in China since 1947 were collected. Genotype identification was carried out using the MLVA15 genotyping protocol based on 15 variable number tandem repeat loci. The genotypes were uniformly numbered and named. The distribution characteristics of the MLVA genotypes of strains were analyzed. Software Bionumerics was used to construct clustering diagrams to analyze the genetic relationships. Results:The MLVA15 clustering analysis subdivided the isolates into 4 major groups and 91 genotypes, 54 of which were unique to China. The genotypes from MLVA15-CHN1 to MLVA15-CHN6 were widely distributed throughout China and in all eras, while other genotypes were restricted to certain regions or eras.Conclusions:This study established a MLVA genotype database of B. anthracis, which provides basis for the understanding of MLVA genetic polymorphisms and the control and molecular source tracing of the anthrax outbreaks in China.

OBJECTIVE To establish the fingerprint of total saponins from Mussaenda pubescens， and to study the spectrum- effect relationship of its hepatoprotective activity. METHODS Ten batches of total saponins from M. pubescens from different origins were prepared using 75% ethanol as solvent. High-performance liquid chromatography （HPLC） and the Similarity Evaluation System for Traditional Chinese Medicine Chromatographic Fingerprints （2012 edition） were used to draw the fingerprints of 10 batches of total saponins from M. pubescens. The similarity evaluation and identification of common peaks were conducted. The same HPLC method was adopted to determine the contents of five triterpenoid saponins （mussaendoside H， mussaendoside U， mussaglaoside C， mussaendoside G and mussaendoside O）. The hepatoprotective effect of total saponins from M. pubescens was investigated by establishing carbon tetrachloride-induced acute liver injury model mice， and the spectrum-effect relationship was studied by using grey correlation analysis. RESULTS There were 11 common peaks in 10 batches of total saponins from M. pubescens， 5 of which were identified， i.e. mussaendoside H （peak 3）， mussaendoside U （peak 7）， mussaglaoside C （peak 8）， mussaendoside G （peak 9） and mussaendoside O （peak 11）； the similarities of 10 batches of samples ranged 0.940- 0.991. Average contents of mussaendoside H， mussaendoside U， mussaglaoside C， mussaendoside G， mussaendoside O were 0.01- 0.05， 0.10-0.21， 0.10-0.18， 0.03-0.08， 0.20-0.40 mg/g， respectively. Ten batches of total saponins from M. pubescens could generally reduce the contents of alanine aminotransferase （ALT） and aspartate aminotransferase （AST） in serum， and tumor necrosis factor-α （TNF-α）， interleukin-6 （IL-6） and IL-1β in liver tissue of model mice （P＜0.05 or P＜0.01）. The E-mail：13878195336@139.com correlation between the common peak areas and the contents of ALT， AST， TNF-α， IL-6 and IL-1β were 0.602-0.757， 0.585-0.833， 0.593-0.795， 0.618-0.820， 0.607-0.804， respectively； the peaks with high correlation were peaks 11， 9 and 8 in order. CONCLUSIONS Ten batches of total saponins from M. pubescens have similar components， and the average contents of mussaendoside H， mussaendoside U， mussaglaoside C， mussaendoside G and mussaendoside O are different. The batches of samples have a certain degree of hepatoprotective effect； mussaendoside O， mussaendoside G and mussaglaoside C may be its main active components.

Objective:To investigate the prediction value of observed to expected lung area to head circumference ratio (o/e LHR), measured at different gestational age with various methods, on indication for extracorporeal membrane oxygenation (ECMO) in fetuses with isolated left-sided congenital diaphragmatic hernia (CDH).Methods:Clinical data of 40 neonates who were diagnosed with left-sided CDH and treated in Guangzhou Women and Children's Medical Center were retrospectively collected from January 2017 to May 2021. The o/e LHRs were prenatally calculated using maximum diameter and tracing method at 22-24 and 31-33 weeks of gestation. According to whether the neonates had indications for ECMO after birth or not, they were divided into ECMO ( n=12) or non-ECMO group ( n=28). Differences in the o/e LHR and general situations between the two groups were analyzed using C hi-square test, independent sample t-test, and non-parametric Mann-Whitney U test. Binary logistic regression was used to analyze the influencing factors for ECMO requirement and receiver operating characteristic (ROC) curve was used to evaluate the value of o/e LHR in predicting the indication for ECMO. Results:Both maximum diameter and tracing method suggested that the o/e LHR at 31-33 gestational weeks was lower than that at 22-24 gestational weeks [maximun diameter method: 40.4 (32.9-51.5) vs 45.1 (36.3-53.4), Z=－2.48, P=0.013; tracing method: 38.6 (33.2-47.6) vs 44.1 (35.9-51.7), Z=－3.29, P=0.001]. There was no statistical difference in o/e LHR detected at the same gestational weeks between the two methods (both P>0.05). Binary logistic regression showed that o/e LHR measured at 31-33 gestational weeks using maximum diameter method was an independent protective factor for ECMO requirement ( OR=0.873, 95% CI: 0.790-0.965, P=0.008). ROC curve analysis showed that the area under the curve for evaluating the predictive value of o/e LHR for ECMO requirement was 0.830 with the sensitivity of 83.3% and the specificity of 71.4% when the cut-off value of o/e LHR at 31-33 gestational weeks was 38.195 measured by maximum diameter method. Conclusions:The o/e LHR measured at 31-33 weeks is lower than that at 22-24 weeks of gestation by both methods. The o/e LHR measured by maximum diameter method at 31-33 weeks of gestation may be useful for predicting the ECMO indication after birth but requiring comprehensive evaluation of clinical conditions due to its insufficient predicting power.

Objective:To evaluate the feasibility and accuracy of a multi-dimensional peripheral quick contrast sensitivity function (pqCSF) measurement established based on Bayesian probability estimation algorithm.Methods:A cross-sectional study was conducted.Nineteen eyes of 12 healthy emmetropic subjects in Zhongshan Ophthalmic Center of Sun Yat-sen University from September 2017 to March 2018 were included, with an average age of (22.92±2.91) years.The average spherical power and cylindrical power were (-0.34±0.52)D and (-0.30±0.42)D, respectively, and the average uncorrected vision acuity was≥1.0.Based on the Bayesian probability algorithm, the peak contrast sensitivity γ max, the peak spatial frequency ? max, the bandwidth β and the low contrast intercept δ were used to quickly describe the contrast sensitivity function (CSF) curve of the full spatial frequency through multi-dimensional pqCSF method.The 16 peripheral visual field positions of all subjects were tested at 6°, 12°, 18° and 24° eccentricity of the superior, inferior, the temporal and nasal visual field by the pqCSF method, but the 18° eccentricity of temporal field, which was near the physiological blind spot, was excluded.The area under Log CSF (AULCSF) of different peripheral visual fields and the Log CSF of 19 spatial frequencies (distributed at equal intervals in logarithmic units) were compared.This study followed the Declaration of Helsinki, and the study protocol was approved by an Ethics Committee of Zhongshan Ophthalmic Center of Sun Yat-sen University (No.2018KYPJ017). Written informed consent was obtained from each subject prior to any examination. Results:With the increase of eccentricity in different visual fields, the AULCSF decreased gradually, and there were significant differences in AULCSF between different eccentricities (all at P<0.05). The AULCSF of the nasal and temporal visual field at 6°, 12° and 24° eccentricity was significantly larger than that of the superior and inferior visual field (all at P<0.05). As the distance from the fovea was increased, the pqCSF, the AULCSF, and the high-frequency cutoff were all decreased, and the standard deviation of AULCSF was increased gradually. Conclusions:The pqCSF method can depict a relatively complete peripheral CSF curve of a wide peripheral visual field, and reflect the function quality of the peripheral vision comprehensively and accurately.

Objective To investigate the effect of piR-9994 on the biological behavior of gastric cancer cells and its possible mechanism. Methods The expression of piR-9994 in gastric cancer cell lines (MGC803 and AGS) and normal gastric epithelial cells (GES-1) were detected by qRT-PCR. MGC803 cell line with piR-9994 overexpression and knockdown were constructed. The effects of piR-9994 expression changes on cell proliferation were detected by MTT and clone formation assay. The scratch wound healing assay and Transwell invasion assay were used to detect cell migration and invasion abilities. qRT-PCR and Western blot were used to detect cell proliferation and EMT-related genes expression. Results The expression level of piR-9994 in MGC803 cells was significantly higher than that in normal gastric epithelial cell line GES-1 (P < 0.05). The overexpression of piR-9994 promoted the proliferation, invasion and metastasis of gastric cancer cells, while piR-9994 knockdown had the opposite effect. piR-9994 expression was closely related to cell cycle, especially in S phase. The overexpression of piR-9994 promoted the expression of proliferation-related genes PCNA, CCND1 and Bcl-2, inhibited the expression of apoptosis gene C-PARP, and promoted the expression of EMT-related genes N-cadherin, MMP7, Twist and Vimentin; while piR-9994 knockdown had the opposite effect. Conclusion Abnormal expression of piR-9994 affects the proliferation, invasion, metastasis and EMT process of gastric cancer cells. piR-9994 may be a new biomarker and therapeutic target for gastric cancer.

Objective: To evaluate the efficacy and safety of laparoscopic surgery in the treatment of gallstones and common bile duct stones. Methods: Eighty-seven patients with gallstones complicated with common bile duct stones who underwent concurrent laparoscopic surgery at Zhoushan Hospital from December 2015 to December 2017 were enrolled.The patients were divided into A group and B group according to the digital table.A group(38 cases) underwent laparoscopic cholecystectomy (LC) combined with laparoscopic common bile duct exploration (LCBDE), and B group(49 cases) underwent endoscopic retrograde cholangiopancreatography (ERCP) and endoscopic sphincterotomy (EST) combined with laparoscopic cholecystectomy (LC). The curative effect of the two groups was observed.The operation time, the success rate of the operation and the rate of laparotomy were recorded in the two groups.The corresponding hospitalization time and cost were compared.The safety of the two different procedures was compared after surgery, and the complications of the two groups were recorded. Results: In A group, the average diameter of common bile duct stones was (1.02±0.25)cm, the average diameter of common bile duct diameter was (1.15±0.25)cm.In B group, the mean diameter of common bile duct stones was (0.99±0.26)cm, and the average diameter of common bile duct was (1.13±0.26) cm.The differences between the two groups were not statistically significant (t=0.513, 0.437, 0.367, P=2.083, 1.533, 1.095). The successful operation rate of A group was 92.11%(35/38), which in B group was 91.84%(45/49), the difference was not statistically significant between the two groups(χ²=0.006, P=0.974). The incidence rate of complications in B group was 20.41%, which was significantly higher than that in A group, the difference was statistically significant(χ²=3.654, P=0.019). The hospitalization time, hospitalization expenses in A group were (10.6±2.6)d, (26 649.8±3 478.6)CNY, respectively, which were significantly better than those in B group (t=21.971, 17.168, all P<0.05). Conclusion The efficacy of LC combined with LCBDE for patients with gallstones complicated with common bile duct stones is better than ERCP/EST combined with LC surgery, and the safety of the former is higher than the latter.

Objective To investigate the effect of preoperative preplacement of nasobiliary drainage in radiofrequency ablation of small hepatocellular carcinoma at hepatic portal. Methods 7 patients with small hepatocellular carcinoma who were admitted to our hospital from June 2014 to June 2017 were selected. All patients were the cases who were prone to injure the bile duct at hepatic portal upon the radiofrequency electrode needle entering liver cancer in the small hepatocellular carcinoma at hepatic portal. The nasal bile duct was pre-placed preoperatively, and infusion of water into nasobiliary bile duct was conducted to expand the bile duct for intraoperative identification and avoiding false puncture. In some cases, after intraoperative bile duct injury, postoperative nasobiliary drainage was given to avoid obstructive jaundice or the occurrence of bile tumor at hepatic portal. Results The treatment effective rate in the patients was 100％ (7/7). Blood biochemistry, routine blood test, liver function were re-examined after surgery. Liver CT or MRI was reexamined 30 to 60 d after surgery. After the preplacement of nasal biliary, there was one case of pancreatitis, with the incidence rate of complications of 14. 3％. The long-term treatment effect was favorable, without tumor recur-rence. Conclusion For small hepatocellular carcinoma at hepatic portal, preoperative preplacment of nasobiliary drainage is used, and the direction of the bile duct during radiofrequency ablation can be clearly observed, so as to effectively avoid the bile duct injury.

Objective To understand the homology of sequence type 562 (ST562) strains of Burkholderia pseudomallei which circulated in two separate continents (Asia and Australia) at different times.Methods Spe Ⅰ restriction fragments and 4-locus multiple locus variable number tandem repeat analysis (MLVA-4) profiles were extracted from MSHR5858 (ST562 Australia strain) and 350105 (ST562 historical strain of Hainan) genomes respectively by in silico analysis and then compared with the PFGE and MLVA-4 results of five ST562 clinical isolates from Hainan to test their homology.Synteny and homology between MSHR5858 and 350105 genomes were evaluated with bioinformatics methods.Results Five ST562 clinical strains from Hainan shared same PFGE pattern (similarity ＞97％) and this pattern coincided to the map of Spe Ⅰ restriction fragments of Australian strain MSHR5858.The amounts of genomic restriction fragments (Spe Ⅰ) for MSHR5858 and 350105 were 31 and 34 respectively,with 31 of them matched by each other.Five ST562 clinical strains of Hainan were distinct by MLVA-4 profiles,among which HPPH43 (MLVA-4 profile:10,8,10,8) was close to Australia strain MSHR5858 (10,8,8,6),containing identical repeat numbers at VNTR loci 2341k and 1788k;while HK003 (11,8,15,7) and HK061 (11,8,17,7) similar to Hainan historical strain 350105 (11,8,11,8),with same repeat numbers at loci 2341k and 1788k also.High-degree synteny and consistency on genomic contents were observed between 350105 and MSHR5858,indicating a similar origin for the 2 strains.Conclusion All inter-continental and historical ST562 strains ofB.pseudomallei had similar genomic characteristics,supporting the assumption that they had a common origin.Also,it is possible that Hainan historical strain 350105 is the ancestor of all circulating ST562 strains.

Objective: To investigate the clinical effect of total cystectomy and W-shaped orthotopic ileal neobladder for bladder cancer. Methods: Total cystectomy and W-shaped orthotopic ileal neobladder were performed on 29 male patients and 3 female patients. Atfer the cystectomy, 35-40 cm terminal ileal segment was isolated to make W-shaped orthotopic ileal neobladder. Bilateral uretero-ileal anastomoses were constructed by nipple technique, urethral stump was sutured to the lowest portion of the ileal pouch. Results: All surgeries were completed successfully. Patients were able to void by increasing the abdominal pressure, daytime continence rate was 87.5% and nighttime continence rate was 78.1%. The mean neobladder capacity was 410.6 mL and the mean postvoid residual urine was22.7 mL at 6 months. After the surgery, ileal neobladder leakage occurred in 3 (9.4%) patients, mild ileus occurred in 2 (6.3%) patients, ileo-urethral anastomosis stricture developed in 1(3.1%) patient, and dysuria occurred in 3 (9.4%) patients because of the mucus.HTe complications were treated properly and satisfactory results were obtained. Erection was preserved in 5 of the 9 male patients with normal erectile function, and normal sexual function was preserved in 1 female patient who underwent uterus-sparing cystectomy. HTe mean follow-up was 15 months.One patient died of distant metastasis and pneumonia, while other patients survived without tumor. Conclusion: Total cystectomy and W-shaped orthotopic ileal neobladder have the advantages of voiding through the native urethra, satisfactory continence, low complication rates of upper urinary tract damage and electrolyte imbalance. W-shaped orthotopic ileal neobladder may be considered as the ifrst option of urinary diversion atfer cystectomy. Postoperative management should focus on complications and follow-up.