The chelating reaction between glutathione peptides and divalent cadmium ion was used as a typical model for investigating the coordination chemistry of SH-containing peptides and heavy metal ions, which was essential to understand the mechanism of intracellular cadmium detoxification.Here, the mutant (CM113R)7 αHL protein nanopore equipped with a first synthesized per-6-quaternary ammonium-β-cyclodextrin (p-QABCD) was used as nanoreactor and detector to investigate the single-molecule reaction of GSH molecules and Cd2+ ions.Different reaction pathways, intermediates, and products could be recognized.Cd2+-GSH reaction was highly dependent on the solution pH.The Cd(GSH)2 was formed at pH 7.4, while the Cd(GSH)2 and Cd2(GSH)2 were formed at pH 9.0.Cd2(GSH)2 was formed by two possible pathways: (1) A Cd2+ ion primarily coordinated with the thiol group of two GSH molecules to form Cd(GSH)2, and then the second Cd2+ ion quickly incorporated with the deprotonated amino group of Cd(GSH)2 to form Cd2(GSH)2;(2) Two Cd2+ ions separately coordinated with the thiol and deprotonated amino group of one GSH molecule to form Cd2(GSH)1, and the second GSH molecule quickly bounded to Cd2+ ions to form Cd2(GSH)2.This work studied the chelating reaction between metal ions and bioactive glutathione at single-molecule level without labeling and chemical modification, and would be important to further understand intracellular mechanisms of detoxification of heavy metals and greatly expand the research field of nanopore single-molecule technique.

ObjectiveTo observe the effect of combined acupuncture-medication anesthesia on visceral function of patients undergone laparoscopic cholecystectomy, and to furtherexplore its clinical and theoretical significance.MethodSixty patients who were going to receive laparoscopic cholecystectomy were enrolled and randomized into an observation group (combined acupuncture-medication anesthesia) and a control group (conventional general anesthesia) by the random number table, 30 subjects in each group. The total bilirubin (TBIL), alanine aminotransferase (ALT), aspertate aminotransferase (AST), urea nitrogen, creatinine, and Mini-Mental State Examination (MMSE) were observed to evaluate the effect of combined acupuncture-medication anesthesia on visceral function of patients undergone laparoscopic cholecystectomy.ResultBefore operation, there were no significant differences in TBIL, ALT, urea nitrogen, creatinine, and MMSE between the observation group and the control group (P>0.05), while the AST level of the observation group was significantly higher than that of the control group before operation(P<0.05). On post-operation day 1, there were no significant differencesin comparing ALT and AST levels between the two groups (P>0.05); the TBIL level in the observation group was significantly lower than that in the control group (P<0.01); there were no significant differences in comparing the urea nitrogen and creatinine levels between the two groups (P>0.05). The MMSE score of the observation group was significantly higher than that of the control group 6 h after the operation (P<0.01). on post-operation day 1, there was no significant difference in comparing MMSE score between the two groups (P>0.05).ConclusionThe combined acupuncture-medication anesthesia and conventional general anesthesia play equal effects on liver and kidney functions in laparoscopic cholecystectomy, thus the two anesthesia methods are both safe andeffective. The combined acupuncture-medication anesthesia has certain protective effects on brain function in early stage, and possibly produces certain protective effectson liver function.

Objective To investigate the patterns for the recurrent and metastatic gastric cancer after curative resection and to indicate the strategy of treatment. Methods 162 patients who had received radical resection and presented post-operation failure during recent 9 years were analyzed. The failure patterns were confirmed by type-B ultrasonic or CT / MRI imaging. 15 of 34 patients with abdominal dropsy were diagnosed by adenocarcinoma cells in the abdominal dropsy. All superficial lymphadens and abdominal wall metastasis were diagnosed via punctuation. 31 patients with gastric remnant and (or) anastomoses recurrence were diagnosed via biopsy. Results Of 162 patients, 63 presented the recurrence or metastasis in multiple sites,including abdominal lymph node (LN) metastases in 46.9 % (76/162), peritoneum metastases in 21.0 % (34/162),gastric remnant and (or) anastomoses recurrence in 19.1% (31/162), liver metastases in 19.1% (31/162), the incidence rates of other parts were all ＜10 %. Meanwhile, out of 76 patients with abdominal lymph node metastases, 48.7 % (37/76) patients with peri-gastric LNs metastases, 31.6 % (24/76) with peri-pancreatic LNs metastases, 19.7 % (15/76) with para-aortic LNs metastases. The incidence of LN metastases was 57.7 % (56/97) in cancer arising from gastric fundus/cardia and 60.4% (29/48) in gastric body and 64.7 % (11/17) in pylorus antrum. Conclusion The regional failure sites for gastric cancer patients with radical resection were dominantly found in the gastric stump/stoma, the peritoneum and pelvic cavity implantation and abdominal cavity LN metastases, especially in the peri-gastric, peri-pancreatic and/or para-aortic LN metastases. The distant place failure sites were mainly in the liver, lung, brain, spondylus, cervical part LN and mediastinal LN metastases. Therefore, we should take chemotherapy, abdominal cavity chemotherapy and regional radiotherapy to prevent the regional district recurrences and distant metastasis after the gastric cancer patients with radical resection. Radiotherapy fields should focus on the gastric stump/stoma and the peri-gastric, peri-pancreatic and para-aortic LN regions.

An improvement of chemiluminescent system for determination of peroxynitrite anion (ONOO－) has been made. In this system, the effect of some antioxidants for scavenging ONOO－ was tested. The constitute of this system and the program of starting were followed as: The ozone (O3) was bubbled through a glass-frit into 10 ml 0.01 mol/L solution of sodium azide in CB(pH 10.5) to generate ONOO－. The 800 μl ozonized solution of azide was injected into a glass tude in situ which contains 100 μl sample and 100 μl luminol solutions to initiate chemiluminescence (CL). The pluses / 6 seconds (CP6S) were determined immediately and continually for 10～30 times. A certain CL intensity (CP6S) was chosen as evaluation index to compare the activity of antioxidants. This chemiluminescent system is sensible, simple and stable. The determination limit was 8.74 μmol/L ONOO－. The linear rang was 8.74～74.04 μmol/L ONOO－. The intra batch and inter batch variation coefficient (CV%) of the analysis were 3.35%(n=10) and 5.52%(n=10) respectively. It was tested that Vit.C, teapolyphenol, procyanidin and thiourea all have effects on scavenging ONOO－.

Objective To determine whether changes in mitral annular spatial conformation and left atrial(LA)volume are closely related to atrial functional mitral regurgitation(AMR)in patients with atrial fibrillation and before or after the sinus rhythm recovery using real-time three-dimensional transesophageal echocardiography(RT-3D TEE).Methods Fifty-five patients with AMR of at least moderate severity who completed one-year of clinical follow-up after ablation were included in this study.Before ablation and after being followed for one year,intercommissural(IC) and anteroposterior(AP) diameter,annular height (AH)and area(MVA),tenting height(TH)and volume(TV),annular spherical index(ASI),fractional area change of MVA(MVA-FAC),and coaptation index(CPI)were defined and measured by mitral valve quantification software(MVQ).Left atrial volume(LAV),left atrial volume index(LAVI),left atrial area (LAA),left atrial diameter(LAD)and effective regurgitant orifice area(EROA)were also recorded. Results After one year of follow-up,AMR decreased significantly in patients with sinus rhythm[EROA:(0.27±0.03)cm2vs(0.21 ±0.04)cm2,P <0.001].LAA,LAV and LAVI were improved in patients with sinus rhythm[LAA:(22.70±3.34)cm2vs(18.80±3.45)cm2,LAV:(52.77±5.41)cm2vs(45.22± 6.49)cm2,LAVI:26.30±3.12 vs 22.98±3.03,all P <0.001].CPI(β=-0.549,P <0.05),MVA-FAC (β=-0.309,P <0.05)and LAV(β=1.712,P <0.05),MVA-FAC)were independently associated with the reduction in EROA.Conclusions AMR can be decreased through the sinus rhythm recovery and maintenance after ablation,which are caused by improvement of left atrial volume load and leaflets coaptation capability.RT-3D TEE may dynamically assess the changes in leaflets/annular configuration during the AMR follow-up.

Objective:To explore the preventive effects of pancreatic duct stent combined with rectal administration of indomethacin suppository for post-endoscopic retrograde cholangiopancreatography pancreatitis (PEP) after difficult bile duct intubation during endoscopic retrograde cholangiopancreatography (ERCP).Methods:From January 2019 to December 2021, patients with biliary and pancreatic diseases undergoing ERCP in Hangzhou Hospital Affiliated to Nanjing Medical University were given 100 mg indomethacin suppository to anal canal 30 minutes before the operation. And those with difficult bile duct intubation during the operation ( n=204) were included in this study. According to the random number table, they were divided into the combination group (implanted with pancreatic duct stent during the operation, n=104) and the indomethacin group (not implanted with stent, n=100). The incidences of hyperamylasemia and PEP were compared between the two groups. Results:The incidences of postoperative hyperamylasemia [21.2% (22/104) VS 34.0% (34/100), χ2=4.22, P=0.040] and PEP [14.4% (15/104) VS 32.0% (32/100), χ2=8.88, P=0.003] in the combination group were significantly lower than those in the indomethacin group. There was no significant difference in the incidence of severe PEP between the two groups [1.0% (1/104) VS 1.0% (1/100), χ2=0.001, P=0.978]. Conclusion:Compared with rectal administration of indomethacin suppository alone, the incidences of hyperamylasemia and PEP after difficult bile duct intubation during ERCP can be further reduced when it is combined with pancreatic duct stent placement.

Circular RNAs (circRNAs), covalently closed continuous RNA loops, are generated from cognate linear RNAs through back splicing events, and alternative splicing events may gener-ate different circRNA isoforms at the same locus. However, the challenges of reconstruction and quantification of alternatively spliced full-length circRNAs remain unresolved. On the basis of the internal structural characteristics of circRNAs, we developed CircAST, a tool to assemble alter-natively spliced circRNA transcripts and estimate their expression by using multiple splice graphs.Simulation studies showed that CircAST correctly assembled the full sequences of circRNAs with a sensitivity of 85.63％-94.32％and a precision of 81.96％-87.55％. By assigning reads to specific iso-forms, CircAST quantified the expression of circRNA isoforms with correlation coefficients of 0.85-0.99 between theoretical and estimated values. We evaluated CircAST on an in-house mouse testis RNA-seq dataset with RNase R treatment for enriching circRNAs and identified 380 cir-cRNAs with full-length sequences different from those of their corresponding cognate linear RNAs. RT-PCR and Sanger sequencing analyses validated 32 out of 37 randomly selected isoforms, thus further indicating the good performance of CircAST, especially for isoforms with low abundance. We also applied CircAST to published experimental data and observed substantial diversity in circular transcripts across samples, thus suggesting that circRNA expression is highly regulated. CircAST can be accessed freely at https://github.com/xiaofengsong/CircAST.

Objective:To investigate outcomes and safety of doxycycline-moxifloxacin sequential regimen in the treatment of Mycoplasma genitalium urethritis/cervicitis. Methods:From June 2019 to December 2020, patients with Mycoplasma genitalium urethritis/cervicitis confirmed by nucleic acid amplification testing were successively recruited at Department of Sexually Transmitted Diseases, Hospital of Dermatology, Chinese Academy of Medical Sciences, and received sequential therapy with oral doxycycline for 7 days followed by oral moxifloxacin for 7 days. Clinical and/or etiological assessment was conducted 2 to 3 weeks after the end of treatment. Fisher′s exact test was used to analyze factors influencing the treatment outcome. Results:Totally, 36 eligible subjects were enrolled, including 30 males and 6 females. Among them, 18 (50%) patients completed post-treatment etiological assessment, which showed that 12 achieved microbiological cure, and treatment failures occurred in 6; another 18 patients achieved clinical cure. The overall response rate to doxycycline-moxifloacin sequential therapy was 83.3% (30/36, 95% confidence interval[ CI]: 70.5%, 96.1%) . The treatment outcome showed no significant association with the patients′ age, gender, marital status, number of sexual partners in the past 1 month, history of sexually transmitted diseases, history of antibiotic use in the past 1 month, or co-infections (all P > 0.05) . Conclusion:The efficacy of doxycycline-moxifloacin sequential regimen is limited in the treatment of Mycoplasma genitalium infections in Nanjing area, and clinicians should be alerted to the possibility of treatment failure in clinical practice.

OBJECTIVE To evaluate the effectiveness， safety， economy， innovation， suitability and accessibility of recombinant Mycobacterium tuberculosis fusion protein （EC）， and to provide evidence for selecting skin detection methods for tuberculosis infection diagnosis and auxiliary diagnosis of tuberculosis. METHODS The effectiveness and safety of EC compared with purified protein derivative of tuberculin （TB-PPD） were analyzed by the method of systematic review. Cost minimization analysis， cost-effectiveness analysis and cost-utility analysis were used to evaluate the short-term economy of EC compared with TB-PPD， and cost-utility analysis was used to evaluate the long-term economy. The evaluation dimensions of innovation， suitability and accessibility were determined by systematic review and improved Delphi expert consultation， and the comprehensive score of EC and TB-PPD in each dimension were calculated by the weight of each indicator. RESULTS The scores of effectiveness， safety， economy， innovation and suitability of EC were all higher than those of TB-PPD. The affordability scores of the two drugs were consistent， while the availability score of EC was lower than those of TB-PPD. After considering dimensions and index weight， the scores of effectiveness， safety， economy， innovation， suitability， accessibility and the comprehensive score of EC were all higher than those of TB-PPD. CONCLUSIONS Compared with TB-PPD， EC performs better in all dimensions of effectiveness， safety， economy， innovation， suitability and accessibility. However， it is worth noting that EC should further improve its availability in the dimension of accessibility.