Objective To compare the clinical efficacy of modified multi-side hole nasobiliary drainage(MHND)via the percuta-neous transhepatic cholangiography(PTC)route with traditional percutaneous transhepatic cholangial drainage(PTCD)for palliative treatment of patients with advanced obstructive cholangiocarcinoma.Methods A retrospective analysis was conducted on the data from 66 patients with advanced cholangiocarcinoma who underwent biliary drainage.Results Both groups normalize temperature and alleviate symptoms of acute cholangitis within 24 h post-puncture.There was no statistically significant difference in laboratory indicators such as white blood cell(WBC),total bilirubin(TBiL),alanine transaminase(ALT),aspartic transaminase(AST)at 48 h post-operation,and in the incidence of bile leakage,biliary peritonitis,and cholangitis during the postoperative hospital stay(P>0.05).However,the incidence of postoperative electrolyte disorders,gastrointestinal symptoms,and the recurrence rate of gastrointestinal symptoms during the follow-up period were significantly higher in the PTCD group compared to the MHND group,while the recur-rence rate of biliary tract infections was slightly higher in the MHND group compared to the PTCD group.The differences between the two groups were statistically significant(P<0.05).Conclusion Modified MHND shows better clinical efficacy in the treatment of patients with advanced tumor jaundice.Compared with traditional PTCD,it not only effectively reduces jaundice and relieves acute cholangitis but also significantly reduces gastrointestinal symptoms during the postoperative period,thereby improving the quality of life for patients.However,it is noteworthy that it may also increase the risk of biliary tract infections.

Background and Aims:Complex choledocholithiasis often coexists with stenosis of the ampulla of Vater,which increases the difficulty and complexity of treatment.If only the stones in the bile duct are removed without addressing the ampullary stenosis,the disease is prone to recurrence.Previously,most treatments involved the use of endoscopic retrograde cholangiopancreatography(ERCP)to guide the wire for sphincterotomy and stone extraction,followed by laparoscopic cholecystectomy.However,ERCP has limitations in handling complex cases.In response,our team pioneered a new method of treating choledocholithiasis combined with stenosis of the ampulla of Vater via a transabdominal approach.This study was performed to investigate the feasibility and efficacy of this method,aiming to provide a new therapeutic option for clinical practice.Methods:A randomized controlled study was conducted with 120 patients treated at Chengdu Second People's Hospital from 2021 to 2023 for gallbladder stones and choledocholithiasis with stenosis of the ampulla of Vater.Patients were divided into an observation group and a control group,with 60 cases in each group.The observation group underwent laparoscopic cholecystectomy with choledochotomy for stone extraction,followed by retrograde guidance of duodenal papillary sphincterotomy through the opened bile duct,simultaneously treating gallbladder,bile duct stones,and stenosis of the ampulla of Vater.The control group underwent traditional ERCP approach for sphincterotomy,stone extraction,and laparoscopic cholecystectomy.Perioperative variables were collected for both groups and the surgical outcomes were compared.Results:Among the 120 patients,54 were male and 66 were female.There were no statistically significant differences between the two groups in terms of stone extraction success rate,intraoperative blood loss,postoperative 24-h total bilirubin,direct bilirubin,transaminases,white blood cell count,jaundice relief time,or incidence rates of bile leakage,retroperitoneal bleeding/infection,and severe pancreatitis(all P>0.05).The observation group had significantly shorter average operative time and postoperative hospital stay compared to the control group(98.67 min vs.110.8 min,P<0.05;3.81 d vs.5.61 d,P<0.05).Additionally,the observation group had a significantly lower incidence of postoperative hyperamylasemia and/or hyperlipasemia and mild pancreatitis(1.67%vs.25.00%,P<0.001;0 vs.10%,P=0.027).Conclusion:The novel transabdominal approach is superior to the ERCP approach in terms of reducing surgery time and hospitalization time,and it carries a lower risk of postoperative mild pancreatitis and hyperamylasemia and/or hyperlipasemia.The stone extraction success rate is comparable to that of ERCP,making it a viable alternative treatment option.

AIM:To investigate the effect of Clostridium butyricum on the intestinal barrier of mice with dex-tran sodium sulfate(DSS)-induced colitis,and to explore its mechanism related to the regulation of endogenous hydrogen sulfide enzymes.METHODS:Forty male C57BL/6J mice(ten per group)were randomly divided into control group,DSS-induced colitis group,and two groups with different doses(1×1010 CFU/L and 1×1011 CFU/L)of Clostridium butyri-cum.To establish the colitis model,mice were given distilled water containing 3.5%DSS.The mice in Clostridium butyri-cum groups received a 10 mL/kg Clostridium butyricum solution by gastric intubation once per day for 14 d.At the end of the experiment,the disease activity index(DAI)and colon length of the mice from each group were recorded.The colon tissue morphology was determined using HE staining,and qRT-PCR was used to measure the mRNA expression levels of two endogenous hydrogen sulfide enzymes,cystathionine β-synthase(CBS)and cystathionine γ-lyase(CSE),and the mu-cus marker LY6/PLAUR domain containing 8(Lypd8).The protein levels of CSE,CBS and macrophage migration inhibi-tory factor(MIF)in the colon were measured by Western blot.Finally,the expression of tight junction proteins,zonula occludens-1(ZO-1)and occludin,in the colon were evaluated by immunohistochemical staining.RESULTS:Compared with control group,the mice in DSS-induced colitis group had a significant increase in DAI and a decrease in colon length(P＜0.05).Furthermore,the DSS-induced colitis mice exhibited loss of colonic mucosal epithelium,disordered crypt ar-rangement,and extensive infiltration of inflammatory cells.Compared with DSS-induced colitis group,both DAI and co-lon shortening were significantly attenuated in the groups with different doses of Clostridium butyricum(P＜0.05).The mice in Clostridium butyricum groups also exhibited relatively intact colonic mucosal epithelial structure with reduced in-flammatory cell infiltration and reversed crypt disorder.The mRNA expression of CSE and CBS was significantly de-creased,and the mRNA expression of Lypd8 was significantly increased in the colon(P＜0.05).Western blot exhibited down-regulation in the protein expression of CSE,CBS and MIF,whereas immunohistochemical staining showed higher ex-pression of ZO-1 and occludin.CONCLUSION:Clostridium butyricum attenuated DSS-induced colitis in mice,mainly through down-regulating the expression of the endogenous hydrogen sulfide enzymes,CSE and CBS,to promote the intesti-nal mucus barrier and protect the integrity of the intestinal epithelium.

Objective To compare the clinical efficacy of modified multi-side hole nasobiliary drainage(MHND)via the percuta-neous transhepatic cholangiography(PTC)route with traditional percutaneous transhepatic cholangial drainage(PTCD)for palliative treatment of patients with advanced obstructive cholangiocarcinoma.Methods A retrospective analysis was conducted on the data from 66 patients with advanced cholangiocarcinoma who underwent biliary drainage.Results Both groups normalize temperature and alleviate symptoms of acute cholangitis within 24 h post-puncture.There was no statistically significant difference in laboratory indicators such as white blood cell(WBC),total bilirubin(TBiL),alanine transaminase(ALT),aspartic transaminase(AST)at 48 h post-operation,and in the incidence of bile leakage,biliary peritonitis,and cholangitis during the postoperative hospital stay(P>0.05).However,the incidence of postoperative electrolyte disorders,gastrointestinal symptoms,and the recurrence rate of gastrointestinal symptoms during the follow-up period were significantly higher in the PTCD group compared to the MHND group,while the recur-rence rate of biliary tract infections was slightly higher in the MHND group compared to the PTCD group.The differences between the two groups were statistically significant(P<0.05).Conclusion Modified MHND shows better clinical efficacy in the treatment of patients with advanced tumor jaundice.Compared with traditional PTCD,it not only effectively reduces jaundice and relieves acute cholangitis but also significantly reduces gastrointestinal symptoms during the postoperative period,thereby improving the quality of life for patients.However,it is noteworthy that it may also increase the risk of biliary tract infections.

Background and Aims:Complex choledocholithiasis often coexists with stenosis of the ampulla of Vater,which increases the difficulty and complexity of treatment.If only the stones in the bile duct are removed without addressing the ampullary stenosis,the disease is prone to recurrence.Previously,most treatments involved the use of endoscopic retrograde cholangiopancreatography(ERCP)to guide the wire for sphincterotomy and stone extraction,followed by laparoscopic cholecystectomy.However,ERCP has limitations in handling complex cases.In response,our team pioneered a new method of treating choledocholithiasis combined with stenosis of the ampulla of Vater via a transabdominal approach.This study was performed to investigate the feasibility and efficacy of this method,aiming to provide a new therapeutic option for clinical practice.Methods:A randomized controlled study was conducted with 120 patients treated at Chengdu Second People's Hospital from 2021 to 2023 for gallbladder stones and choledocholithiasis with stenosis of the ampulla of Vater.Patients were divided into an observation group and a control group,with 60 cases in each group.The observation group underwent laparoscopic cholecystectomy with choledochotomy for stone extraction,followed by retrograde guidance of duodenal papillary sphincterotomy through the opened bile duct,simultaneously treating gallbladder,bile duct stones,and stenosis of the ampulla of Vater.The control group underwent traditional ERCP approach for sphincterotomy,stone extraction,and laparoscopic cholecystectomy.Perioperative variables were collected for both groups and the surgical outcomes were compared.Results:Among the 120 patients,54 were male and 66 were female.There were no statistically significant differences between the two groups in terms of stone extraction success rate,intraoperative blood loss,postoperative 24-h total bilirubin,direct bilirubin,transaminases,white blood cell count,jaundice relief time,or incidence rates of bile leakage,retroperitoneal bleeding/infection,and severe pancreatitis(all P>0.05).The observation group had significantly shorter average operative time and postoperative hospital stay compared to the control group(98.67 min vs.110.8 min,P<0.05;3.81 d vs.5.61 d,P<0.05).Additionally,the observation group had a significantly lower incidence of postoperative hyperamylasemia and/or hyperlipasemia and mild pancreatitis(1.67%vs.25.00%,P<0.001;0 vs.10%,P=0.027).Conclusion:The novel transabdominal approach is superior to the ERCP approach in terms of reducing surgery time and hospitalization time,and it carries a lower risk of postoperative mild pancreatitis and hyperamylasemia and/or hyperlipasemia.The stone extraction success rate is comparable to that of ERCP,making it a viable alternative treatment option.

AIM:To investigate the effect of Clostridium butyricum on the intestinal barrier of mice with dex-tran sodium sulfate(DSS)-induced colitis,and to explore its mechanism related to the regulation of endogenous hydrogen sulfide enzymes.METHODS:Forty male C57BL/6J mice(ten per group)were randomly divided into control group,DSS-induced colitis group,and two groups with different doses(1×1010 CFU/L and 1×1011 CFU/L)of Clostridium butyri-cum.To establish the colitis model,mice were given distilled water containing 3.5%DSS.The mice in Clostridium butyri-cum groups received a 10 mL/kg Clostridium butyricum solution by gastric intubation once per day for 14 d.At the end of the experiment,the disease activity index(DAI)and colon length of the mice from each group were recorded.The colon tissue morphology was determined using HE staining,and qRT-PCR was used to measure the mRNA expression levels of two endogenous hydrogen sulfide enzymes,cystathionine β-synthase(CBS)and cystathionine γ-lyase(CSE),and the mu-cus marker LY6/PLAUR domain containing 8(Lypd8).The protein levels of CSE,CBS and macrophage migration inhibi-tory factor(MIF)in the colon were measured by Western blot.Finally,the expression of tight junction proteins,zonula occludens-1(ZO-1)and occludin,in the colon were evaluated by immunohistochemical staining.RESULTS:Compared with control group,the mice in DSS-induced colitis group had a significant increase in DAI and a decrease in colon length(P＜0.05).Furthermore,the DSS-induced colitis mice exhibited loss of colonic mucosal epithelium,disordered crypt ar-rangement,and extensive infiltration of inflammatory cells.Compared with DSS-induced colitis group,both DAI and co-lon shortening were significantly attenuated in the groups with different doses of Clostridium butyricum(P＜0.05).The mice in Clostridium butyricum groups also exhibited relatively intact colonic mucosal epithelial structure with reduced in-flammatory cell infiltration and reversed crypt disorder.The mRNA expression of CSE and CBS was significantly de-creased,and the mRNA expression of Lypd8 was significantly increased in the colon(P＜0.05).Western blot exhibited down-regulation in the protein expression of CSE,CBS and MIF,whereas immunohistochemical staining showed higher ex-pression of ZO-1 and occludin.CONCLUSION:Clostridium butyricum attenuated DSS-induced colitis in mice,mainly through down-regulating the expression of the endogenous hydrogen sulfide enzymes,CSE and CBS,to promote the intesti-nal mucus barrier and protect the integrity of the intestinal epithelium.

ObjectiveTo establish an ischemia-reperfusion model in cynomolgus macaques and to analyse the effects of edaravone intervention. MethodsA total of fifteen adult male cynomolgus macaques were randomly divided into three groups: sham operation (Sham group, n=3), ischemia-reperfusion model (Model group, n=6) and edaravone treatment (Edaravone group, n=6). Ischemic-reperfusion model of cynomolgus macaques was established by clamping the M1 branch of the left cerebral artery for 1 h. After 2 h of reperfusion, the animals in Edaravone group were injected with 0.5 mg/kg edaravone intravenously for intervention treatment, while the animals in Sham and Model groups were injected with an equal volume of normal saline intravenously, twice a day, from the 2nd to 7th day. The behavioral video recordings, clinical observations and neurological deficit scores of cynomolgus macaques were obtained, and brain edema volume and cerebral ischemia volume were statistically analyzed. ResultsCompared with the Sham group, the animals in Model group showed typical symptoms of ischemic stroke, with a significant increase in the neurological deficit score， the volumes of edema and infarct of brain tissue (all P＜0.01). Compared with Model group, the neurological deficit score, the volumes of edema and infarct of brain tissue were significantly reduced in Edaravone group (all P＜0.05). ConclusionAn animal model of ischemia-reperfusion in cynomolgus macaques was successfully established, and edaravone was confirmed to alleviate the damage caused by ischemia-reperfusion.

In order to develop a simple and efficient site-directed mutagenesis solution, the Gibson assembly technique was used to clone the cyclin dependent kinase 4 gene with single or double site mutations, with the aim to simplify the overlap extension PCR. The gene fragments containing site mutations were amplified using a strategy similar to overlap extension PCR. Meanwhile, an empty plasmid was digested by double restriction endonucleases to generate a linearized vector with a short adaptor overlapping with the targeted gene fragments. The gene fragments were directly spliced with the linearized vector by Gibson assembly in an isothermal, single-reaction, creating a recombinant plasmid. After the recombinant plasmids were transformed into competent Escherichia coli DH5α, several clones were screened from each group. Through restriction analysis and DNA sequencing, it was found that the randomly selected clones were 100% target mutants. Since there was neither tedious multiple-round PCR amplification nor frequent DNA extraction operation, and there was no need to digest the original plasmid, this protocol circumvents many factors that may interfere with the conventional site-directed mutagenesis. Hence, genes with single or multiple mutations could be cloned easily and efficiently. In summary, the major defects associated with overlap extension PCR and rolling circle amplification were circumvented in this protocol, making it a good solution for site-directed mutagenesis.
Clone Cells ; Mutagenesis, Site-Directed ; Mutation ; Plasmids/genetics* ; Polymerase Chain Reaction/methods*

A 75-year-old male patient received an IV infusion of lappaconitine hydrobromide for injection 4 mg dissolved into 5% glucose injection 250 ml once daily due to bone metastases of lung cancer. After a single time of lappaconitine treatment, the patient developed muscle tremor mainly in the limbs, which was obvious in the sitting and standing positions and disappeared after lying flat. After 2 times of lappaconitine treatment, the muscle tremor was aggravated and he was unable to take care of himself due to tremor, showing that his hands could not hold things and walking was difficult. No other abnormal signs were found except the increase of muscle tension. The extrapyramidal symptoms (Parkinson-like symptoms) induced by lappaconitine was considered. Then lappaconitine was stopped and other treatments continued. About 24 hours of lappaconitine withdrawal, the patient′s muscle tremor disappeared. After 8 days of observation, his symptoms did not recur.

A 56-year-old male patient with advanced hepatocellular carcinoma received targeted therapy with lenvatinib 12 mg once daily. After 30 days of medication, the patient developed epistaxis without predisposing factors in the morning, which occurred once every half a month and could be stopped after compression. The drug was not stopped. After 97 days of medication, the patient developed abdominal pain. Physical examination showed abdominal tenderness, especially in the upper and middle abdomen, rebound pain, and muscle tension. Abdominal CT examination showed pneumoperitoneum, hydrops, and perforation of hollow organs of abdomen. Lenvatinib was withdrawn and the conservative treatments with fasting, gastrointestinal decompression, anti-infection, acid suppression, and rehydration were given. After 22 days of treatments, his abdominal pain was relieved and the fluid diet was restored. After 26 days of treatments, the patient had exhaust and defecation. After that, the patient took lenvatinib with the same dose as before by himself again for one week, and epistaxis recurred. There was no epistaxis after discontinuation of lenvatinib again. It was considered that epistaxis and perforation of hollow organs might be related to lenvatinib.