ObjectiveTo observe the effect of donor liver pretreated by breviscapine on liver transplantation ischemia/reperfusion injury in rats. Methods SD rats served as liver donors and recipients (n =48 each).The recipients were divided into four groups by random number table.The donors in groups A and C were not pretreated with breviscapine,but those in groups B and D were pretreated with 20 mg/L Breviscapine.The cold ischemia time in donor livers of groups A and B was 30-40 min,and that in groups C and D was 12 h. Clotting function, liver function, serum thrombomodulin,caspase3,and relative activity of NF-kB after liver transplantation were assessed,and the pathological changes and TUNEL apoptosis staining were observed.ResultsThe mortality in groups C and D was 40.0％ (8/20) and 29.4％ (5/17),respectively (P＞0.05).There were no significant changes in coagulation function in all groups after operation. The liver function was improved,pathological lesions were alleviated,and apoptosis rate,serum TM,caspase3 expression and activity of NF-kB in the liver tissues of group D were significantly decreased as compared with group C at 3rd day after operation (P＜0.01),but all these parameters in group B had no significant change compared to group A.ConclusionPretreatment of donor livers with breviscapine can reduce the ischemia/reperfusion injury and apoptosis after liver transplantation in rats probably by inhibiting the apoptosis-related pathway and alleviating the damage to the endothelial cells of the liver microcirculation.

Objective To explore the application of a three-dimensional simulation system in extended liver resections.Methods A three-dimensional surgical simulation system was used for preoperative assessment and for computer simulation to estimate the resected liver volume,the residual liver volume,and the surgical resection margins.The software virtual liver resection was used to choose the most optimal surgical option.The actual resection,the postoperative liver volume and the actual surgical resection margins were compared.Results 1 patient was diagnosed by ultrasound to develop ascites on post-operative day 2.Another patient was diagnosed by transthoracic ultrasound to have moderate pleural effusion on postoperative day 1.Postoperative liver failure developed in 1 patient.There was no patient who developed bile leakage.The average length of stay was 6 ～ 85 days (average 22 d).There was no recurrence within 1 month after surgery and there was no postoperative deaths.Condusions The three-dimensional surgery simulation system was effective.It gave an accurate evaluation and simulation of the liver surgery.In extended liver resection,it gave good guidance and help.

Acute gastrointestinal dysfunction is a common and important complication of sepsis. As no exiting formal definition and classification of gastrointestinal dysfunction, most of the treatment strategies for gastrointestinal dysfunction are not based on clinical evidence, but on their own clinical experience. Experts of traditional Chinese medicine, integrated traditional Chinese and Western medicine and Western medicine from various disciplines in Shanghai are organized by the Shanghai Society of Integrated Traditional Chinese and Western Medicine and the Emergency Department Branch of Shanghai Physicians Association. After repeated discussion, literature search and formulation of the outline, we developed consensus on gastrointestinal dysfunction secondary to sepsis with integrating Traditional Chinese Medicine and Western medicine by consulting extensively on clinical experts in the fields of emergency medicine, gastroenterology, general surgery, infectious medicine and traditional Chinese medicine, and holding several expert forums and consultation meetings. This clinical expert consensus focused on acute gastrointestinal injury (AGI) classification and inducer of sepsis. In this consensus, the common symptoms, diagnosis, classifications, treatment strategies and suggestions of acute gastrointestinal injury or dysfunction secondary to sepsis were explored from the aspect of both Traditional Chinese Medicine and Western medicine.

Objective To investigate a anatomy research and clinic application of defect of thumb by reverse dorsoradial thumb flap of different rotation point.Methods The origin,course,distribution and vascular chain of the first metacarpal dorsoradial artery of thumb from 11 adult cadaveric hand specimens perfused by red latex were explored from September,2012 to December,2016.There were 3 different rotation points:the proximal of the metacarpophalangeal joint,proximal basal of the first proximal phalanx and distal of the first proximal phalanx.Each could be used as reverse flow flap to repair the defect of thumb.Results The first metacarpal dorsoradial artery of thumb originated from the radial artery and the initial diameter was (0.68±0.26) mm,diagonally across the extensor pollicis brevis tendon and then along the radialis part and terminated in the proximal of the first proximal phalanx of the vascular chain.There was a constant communicating branch among the proximal metacarpophalangeal joint,proximal basal of the first proximal phalanx and digital arterial dorsal branch.All the proximal of the metacarpophalangeal joint 4.3 mm to 10.2 mm and proximal basal of the first proximal phalanx 4.9 mm to 7.2 mm could be used as the rotation point of the flap.The flap of the first promixal phalanx blood supply was based on the vascular chain of neurocutaneous.There was a constant dorsal branch of the pollical artery,which was 8.6 mm to 10.3 mm far from the interphalangeal joint,could be used as the rotation point of the flap.Twenty-four cases with soft tissue defects of thumb were repaired by reverse dorsoradial flap.The flap size ranged from 2.0 cm×1.5 cm to 3.5 cm×2.8 cm.The follow-up period was 3 months to 12 months,protective sensations were restored,and skin flap two-point discrimination were 9.0 mm to 12.0 mm.The appearance of the thumbs was satisfactory.Conclusion Different rotation point of reverse dorsoradial flap can successfully repair the defect of thumb.The operation has advantages of simple,reliable blood supply,high success rate and is an ideal option for reconstruction the defect of thumb.

Objective To construct rat lysine-specific histone demethylase 1 (LSD1) overexpression plasmids and LSD 1 demethylation fragment disfunction plasmids,and to evaluate their expression levels in HEK293T cells.Methods LSD1 fragments were amplified by PCR,and LSD1 demethylation disfunction fragments were amplified by overlap PCR.Rat-specific LSD1 overexpression plasmids and LSD1 demethylation disfunction plasmids were constructed and verified by agarose gel electrophoresis and sequencing.HEK293T cells were infected using the validated recombinant plasmids and blank vectors,and the stably expressed cells were selected by puromycin.The expression of LSD1 in the stably expressed HEK293T was detected by Western Blot and real-time quantitative PCR.Results The results of agarose gel electrophoresis and sequencing showed that LSD1 overexpression plasmids and LSD1 demethylation disfunction plasmids were successfully constructed.The Western Blot and real-time quantitative PCR results showed that compared with the blank group,the relative expression of LSD1 and mRNA in the LSD1 overexpression group and the LSD1 demethylation disfunction group were up-regulated,and the differences were statistically significant(all P＜0.01).Conclusions The constructed LSD1 overexpression plasmids and LSD1 demethylationi disfunction plasmids can achieve overexpression of LSD1 gene in HEK293T cells.This paper lays a foundation for further study of the relationship between LSD 1 gene and related diseases and demethylation of LSD 1.

Objective:To investigate the clinical difference between primary suture and T tube drainage in laparoscopic choledocholithotomy.Methods:The clinical data of 124 patients treated by laparoscopic choledocholithotomy in Suzhou Municipal Hospital from December 2018 to February 2020 were retrospectively studied. The patients were divided into the primary suture group (71 cases) and the T tube drainage group (53 cases) according to the different surgical methods, and the differences in the relevant treatment indicators were compared between the two groups.Results:There were no statistically significant differences between the two groups in gender, hypertension, diabetes mellitus, preoperative aspartate aminotransferase, preoperative alanine aminotransferase, preoperative total bilirubin, preoperative common bile duct diameter, postoperative length of stay, total cost of hospitalization, postoperative exhaust time, or postoperative biliary leakage, et al. Compared with the T tube drainage group, the primary suture group had more single choledocholithiasis before operation (33 cases vs. 15 cases), shorter operation time: (100.14 ± 38.90) h vs. (140.45 ± 54.17) h, less intraoperative bleeding: (35.70 ± 30.17) ml vs. (49.53 ± 34.58) ml, and later extraction time of Winslow hole drainage tube after operation: (7.15 ± 2.61) d vs. (5.45 ± 3.35) d, and the differences were statistically significant ( P<0.05). Conclusions:Under the condition of strictly controlling the indications of primary suture and being operated by general surgeons who can skillfully operate laparoscope and choledochoscope, laparoscopic choledocholithotomy for primary suture has better curative effect than T tube drainage, and has higher clinical application value.

Objective To construct a human renal epithelial cell line HEK293T by CRISPR-Cas9-based site-directed knock-in of vascular endothelial growth factor 165 (VEGF165) gene, and avoid the off-target effect caused by lentivirus infection. Methods The VEGF165 expression vector with homologous arm (pUCm-T-VEGF165 plasmid) and the sgRNA expression vector [pSpCas9(BB)-2A-Puro-sgRNA plasmid] were designed and constructed based on the DNA sequence of the EZH2 gene, and then co-transfected into HEK293T cells. The expression of VEGF165 mRNA was detected by qPCR and the expressions of VEGF165 proteins were detected by Western Blot. Results The qPCR and Western Blot results showed that, comparing with the control, the pUCm-T-VEGF165 plasmid and pSpCas9(BB)-2A-Puro-sgRNA plasmid, the expression of the co-transfection plasmid were significantly increased, i.e. 3.42±0.30 vs. 1.02±0.21, 1.13±0.16 and 0.98±0.18 for the VEGF165 mRNA level (all P<0.01), and 1.13±0.16 vs. 1.02±0.06, 0.88±0.03 and 0.80±0.05 for the VEGF165 protein level (all P<0.01), respectively. Besides, the expression of EZH2 was significantly down-regulated, i.e. 0.14±0.06 vs. 1.08±0.11, 1.02±0.12 and 1.13±0.16 for the EZH2 mRNA level (all P<0.01), and 0.23±0.03 vs. 1.05±0.13, 0.91±0.04 and 0.81±0.06 for the EZH2 protein level (all P<0.01), respectively. This result showed that the VEGF165 was successfully inserted into the EZH2 genome, interfering the EZH2 expression. Conclusions VEGF165 gene can be successfully knocked into HEK293T cells by CRISPR/Cas9 system.

Objective:To study the characteristics and influencing factors of vaginal microbiota in normal pregnant women.Methods:This study was based on a cohort of pregnant women established in Anqing Municipal Hospital Affiliated to Anhui Medical University from February 2018 to February 2020. Vaginal samples of normal pregnant women who met the inclusion and exclusion criteria were ordered by the gestational weeks at sampling. Five samples were randomly selected from each gestational week group and if the samples were less than five, all samples were included. Sequencing of the V3-V4 region of the 16S rRNA gene was performed. Dominant species were analyzed by MicrobiomeAnalyst. Alpha diversity was measured with Chao1, Observed Features, Shannon diversity, Simpson diversity, Faith_pd and Pielou′s Evenness. The dominant status of Lactobacillus was also described and compared. Multiple linear regression and logistic regression were used to analyze the factors influencing vaginal microbiota. Analysis of variance and Kruskal Wallis test were used for statistical analysis of continuous variables, and Chi-square test and Fisher′s exact test were used for categorical data. The differences were considered statistically significant when the P value was less than 0.05. Results:This study enrolled 91 pregnant women (91 vaginal samples) with an average age of (27.37±3.60) years. There were 18, 56 and 17 vaginal samples collected at the median gestational age of 11.93 weeks (the first trimester), 19.43 weeks (the second trimester) and 38.29 weeks (the third trimester), respectively. The relative abundance of Firmicutes and Lactobacillus was 91.30% and 87.67%, respectively. Lactobacillus iners and Lactobacillus crispatus had a relative abundance of 43.95% and 36.33%, respectively. Moreover, Lactobacillus iners-dominated vaginal microbiota was detected in all trimesters. The number of samples with high relative abundance of Lactobacillus iners gradually decreased with gestational age. Lactobacillus crispatus-dominated vaginal microbiota was found in the second and third trimesters and the number of samples with high relative abundance gradually increased during pregnancy. The Alpha diversity of vaginal microbiota had a decreasing trend during the gestation. There were significant differences in Pielou′s Evenness diversity index of vaginal microbiota between different smoking groups ( P<0.05) and in Shannon diversity index between different drinking groups ( P<0.05). There were significant differences in Chao1, Observed Features and Faith_pd diversity index of vaginal microbiota between pregnant women with different education ( P<0.05) and in Shannon and Simpson diversity index between different income groups ( P<0.05). Conclusions:Vaginal microbiota was dominated by Lactobacillus in normal pregnant women. The dominance of Lactobacillus iners gradually decreased, while that of Lactobacillus crispatus increased during gestation. In normal pregnant women, the Alpha diversity of vaginal microbiota was correlated with smoking, drinking, education and family annual income. Smoking cessation and drinking before pregnancy were related to lower Alpha diversity of vaginal microbiota in pregnant women, while lower education and higher family income were associated with higher Alpha diversity.

OBJECTIVE To establish the method for monitoring the dynamic changes of odor components in Cornus officinalis during processing . METHODS The decoction pieces of C. officinalis with different processing time were prepared by the wine steaming method . The dynamic changes of odor components were obtained by using ultra -fast gas electronic nose ；odor components were identified by comparing with AroChemBase database ；the dynamic changes of odor compounds were analyzed in combination with peak area ，and the chemical pattern recognition analysis were carried out . RESULTS A total of 12 common peaks of odor components were identified in the fingerprints of raw C. officinalis，and 21 in the fingerprints of decoction pieces of C. officinalis. Eight odor components with the high proportion of peak area during processing were ethanol ， isopropyl alcohol ， 2- methylpropylaldehyde，ethyl acetate ，2-methylbutanal，isoamyl alcohol ，2-hexanol and furfural ，among which ，the peak areas of ethanol，isoamyl alcohol and 2-hexanol showed a trend of first increasing and then decreasing ；at 24 h of processing ，their peak areas were still higher than those of raw products . The peak areas of ethyl acetate ，2-methylbutanal and furfural nearly increased with the increase of processing time . Variable importance in projection of above eight odor components were all greater than 1. CONCLUSIONS The method is established for monitoring the dynamic changes of odor components of C. officinalis during processing. Eight odor components such as ethanol can be used as monitoring indicators of C. officinalis dring processing .

Objective To establish bovine corneal opacity and permeability （BCOP） test， and determine its predictive ability for the eye irritation evaluation of cosmetics. Methods A total of ten reference chemicals were selected to establish the BCOP test. Then eye irritation of 16 routinely collected cosmetics in our laboratory was predicted. In vitro scores were calculated by the change in the opacity and sodium fluorescein permeability after exposure to the testing cosmetics， and subsequently compared with the historical data by Draize test. Results Reference chemicals with known irritation classification were correctly classified by the BCOP test， which was consistent with the classification of UN globally harmonized system of classification and labeling of chemicals. Moreover， the specificity of the BCOP test for the classification of non-irritating cosmetics samples was 80.0% （8/10）， and the sensitivity for weak to mild irritating cosmetics samples was 83.3% （5/6）. The BCOP test demonstrated an overall classification consistency of 81.3% （13/16） with in vivo test. Conclusion BCOP test may be independently used to identify chemicals with potential eye irritation and serious eye damage， suggesting it is significant for in vitro integrated test strategy for predicting eye irritation due to cosmetics.