This studyobtained the proteins of virulence factors EsxA and EsxB from Mycobacterium haemophilum and explored their interactions both in vitro and in vivo.The aim was to lay a foundation for further analysis of the functional mechanisms of EsxA and EsxB,and to further the development of drugs targeting Mycobacterium haemophilum.On the basis of bioinformatics analysis of the virulence factors EsxA and EsxB from Mycobacterium haemophilum,we performed molecular cloning,using Mycobacterium haemophi-lum as a template to construct recombinant plasmids EsxA-pGl01,EsxB-pGl01,and EsxB-pET-29b.The proteins of EsxA,EsxB,and their complex were expressed with a prokaryotic system,then purified through nickel-affinity chromatography and gel filtration chromatography.Intracellular colocalization was determined through fluorescence colocalization.Prokaryotic expression systems for EsxA,EsxB,and their complex were successfully constructed,and purified proteins were obtained.Fluorescence colocalization indi-cated that EsxA and EsxB interacted in vivo.In vivo fluorescence colocalization and in vitro complex formation suggested that EsxA and EsxB interacted both intracellularly and extracellularly.Our findings lay a foundation for studying the pathogenic mechanisms of the virulence factors EsxA and EsxB in Mycobacterium haemophilum,and performing structural analysis of their complex.

This studyobtained the proteins of virulence factors EsxA and EsxB from Mycobacterium haemophilum and explored their interactions both in vitro and in vivo.The aim was to lay a foundation for further analysis of the functional mechanisms of EsxA and EsxB,and to further the development of drugs targeting Mycobacterium haemophilum.On the basis of bioinformatics analysis of the virulence factors EsxA and EsxB from Mycobacterium haemophilum,we performed molecular cloning,using Mycobacterium haemophi-lum as a template to construct recombinant plasmids EsxA-pGl01,EsxB-pGl01,and EsxB-pET-29b.The proteins of EsxA,EsxB,and their complex were expressed with a prokaryotic system,then purified through nickel-affinity chromatography and gel filtration chromatography.Intracellular colocalization was determined through fluorescence colocalization.Prokaryotic expression systems for EsxA,EsxB,and their complex were successfully constructed,and purified proteins were obtained.Fluorescence colocalization indi-cated that EsxA and EsxB interacted in vivo.In vivo fluorescence colocalization and in vitro complex formation suggested that EsxA and EsxB interacted both intracellularly and extracellularly.Our findings lay a foundation for studying the pathogenic mechanisms of the virulence factors EsxA and EsxB in Mycobacterium haemophilum,and performing structural analysis of their complex.

Objective To investigate the effect of dihydroartemisinin(DHA)for enhancing the inhibitory effect of cisplatin(DDP)on DDP-resistant nasopharyngeal carcinoma cell line HNE1/DDP and explore the mechanism.Methods CCK-8 method was used to assess the survival rate of HNE1/DDP cells treated with DHA(0,5,10,20,40,80,and 160 μmol/L)and DDP(0,4,8,16,32,64,128 μmol/L)for 24 or 48 h,and the combination index of DHA and DDP was calculated using Compusyn software.HNE1/DDP cells treated with DHA,DDP,or their combination for 24 h were examined for cell viability,proliferation and colony formation ability using CCK-8,EdU and colony-forming assays.Flow cytometry was used to detect cell apoptosis and intracellular reactive oxygen species(ROS).The expression levels of apoptosis-related proteins cleaved PARP,cleaved caspase-9 and cleaved caspase-3 were detected by Western blotting.The effects of N-acetyl-cysteine(a ROS inhibitor)on proliferation and apoptosis of HNE1/DDP cells with combined treatment with DHA and DDP were analyzed.Results Different concentrations of DHA and DDP alone both significantly inhibited the viability of HNE1/DDP cells.The combination index of DHA(5 μmol/L)combined with DDP(8,16,32,64,128 μmol/L)were all below 1.Compared with DHA or DDP alone,their combined treatment more potently decreased the cell viability,colony-forming ability and the number of EdU-positive cells,and significantly increased the apoptotic rate,intracellular ROS level,and the expression levels of cleaved PARP,cleaved caspase-9 and cleaved caspase-3 in HNE1/DDP cells.N-acetyl-cysteine pretreatment obviously attenuated the inhibitory effect on proliferation and apoptosis-inducing effect of DHA combined with DDP in HNE1/DDP cells(P<0.01).Conclusion DHA enhances the growth-inhibitory and apoptosis-inducing effect of DDP on HNE1/DDP cells possibly by promoting accumulation of intracellular ROS.

Objective:To analyze the intraoperative hemodynamic management by hypotension prediction index(HPI)in one patient underwent robot-assisted laparoscopic cystectomy,and to provide the reference for anesthesia monitoring and hemodynamic management in the similar major surgery.Methods:The clinical data,intraoperative hemodynamic data,usage and dosage of vasoactive drugs,and clinical outcomes of one patient underwent robot-assisted laparoscopic cystectomy with HPI-guided intraoperative hemodynamic management were retrospectively analyzed,and the relevant literatures were reviewed.Results:The patient,a 72-year-old female,was admitted due to macroscopic hematuria for 5 months accompanied by dysuria for 3 months.The cystoscope results showed a 7 cm× 7 cm× 5 cm mass on the right side of the bladder trigone and a 4 cm × 3 cm × 3 cm mass near the bladder neck.The positron emission tomography/computed tomography(PET/CT)results showed thickening of the right posterior bladder wall with high metabolism,and the preliminary diagnosis was bladder malignancy.After preoperative anesthesia evaluation,the robot-assisted laparoscopic cystectomy was planned.After entering the operating room,the routine monitoring was conducted,and the monitor equipped with HPI software was used to guide intraoperative hemodynamic management.After routine anesthesia induction,the tracheal intubation was performed by video laryngoscope.The patient experienced intraoperative hypotension(IOH)for six times,the cumulative time of mean arterial pressure(MAP)＜65 mmHg was 13.7 min,accounting for 4.40％of the anesthesia duration,and the time-weighted average of MAP＜65 mmHg was 0.28 mmHg.The time range with HPI≥85 roughly overlapped with and included the period of MAP＜65 mmHg.At 146 time points with HPI≥85,the MAP remained greater than 65 mmHg at 68.5％(100/146)of the points.At 47 time points with MAP＜65 mmHg,HPI≥85 occurred at 97.9％(46/47)of the points.On the first postoperative day,the patient's hypersensitive cardiac troponin I was＜0.01 μg·L-1,and no perioperative adverse events occurred.The patient was discharged on the eighth day.Conclusion:HPI can promptly and accurately predict the occurrence of IOH in the patients undergoing robot-assisted laparoscopic cystectomy.The use of HPI-based hypotension correction strategies during surgery can maintain the time-weighted average of MAP＜65 mmHg at a lower level.

Pancreatic cancer is a highly malignant digestive tract tumor with hidden symptoms,limited treatment options and rapid progression.With an increasing incidence rate year by year,pancreatic cancer has increasingly become a prominent issue endangering public health,causing a huge social burden.Although there was no significant improvement in survival rates for pancreatic cancer patients in the past two decades,recent progress in epidemiology,basic research and clinical research of pancreatic cancer has accelerated significantly compared to the past.Some findings have already enabled a small proportion of pancreatic cancer patients to achieve better survival.This article provided a review of the significant progress made in research,diagnosis and treatment of pancreatic cancer in 2023.

Objective To investigate the effect of dihydroartemisinin(DHA)for enhancing the inhibitory effect of cisplatin(DDP)on DDP-resistant nasopharyngeal carcinoma cell line HNE1/DDP and explore the mechanism.Methods CCK-8 method was used to assess the survival rate of HNE1/DDP cells treated with DHA(0,5,10,20,40,80,and 160 μmol/L)and DDP(0,4,8,16,32,64,128 μmol/L)for 24 or 48 h,and the combination index of DHA and DDP was calculated using Compusyn software.HNE1/DDP cells treated with DHA,DDP,or their combination for 24 h were examined for cell viability,proliferation and colony formation ability using CCK-8,EdU and colony-forming assays.Flow cytometry was used to detect cell apoptosis and intracellular reactive oxygen species(ROS).The expression levels of apoptosis-related proteins cleaved PARP,cleaved caspase-9 and cleaved caspase-3 were detected by Western blotting.The effects of N-acetyl-cysteine(a ROS inhibitor)on proliferation and apoptosis of HNE1/DDP cells with combined treatment with DHA and DDP were analyzed.Results Different concentrations of DHA and DDP alone both significantly inhibited the viability of HNE1/DDP cells.The combination index of DHA(5 μmol/L)combined with DDP(8,16,32,64,128 μmol/L)were all below 1.Compared with DHA or DDP alone,their combined treatment more potently decreased the cell viability,colony-forming ability and the number of EdU-positive cells,and significantly increased the apoptotic rate,intracellular ROS level,and the expression levels of cleaved PARP,cleaved caspase-9 and cleaved caspase-3 in HNE1/DDP cells.N-acetyl-cysteine pretreatment obviously attenuated the inhibitory effect on proliferation and apoptosis-inducing effect of DHA combined with DDP in HNE1/DDP cells(P<0.01).Conclusion DHA enhances the growth-inhibitory and apoptosis-inducing effect of DDP on HNE1/DDP cells possibly by promoting accumulation of intracellular ROS.

Background: Postoperative pain management is crucial for patients undergoing total knee arthroplasty (TKA). There have been many recent clinical trials on post-TKA peripheral nerve block; however, they have reported inconsistent findings. In this meta-analysis, we aimed to comprehensively analyze studies on post-TKA analgesia to provide evidence-based clinical suggestions. Methods: We performed a computer-based query of PubMed, Embase, the Cochrane Library, and the Web of Science to retrieve related articles using neurothe following search terms: nerve block, nerve blockade, chemodenervation, chemical neurolysis, peridural block, epidural anesthesia, extradural anesthesia, total knee arthroplasty, total knee replacement, partial knee replacement, and others. After quality evaluation and data extraction, we analyzed the complications, visual analogue scale (VAS) score, patient satisfaction, perioperative opioid dosage, and rehabilitation indices. Evidence was rated using the Grading of Recommendations Assessment, Development, and Evaluation approach. Results: We included 16 randomized controlled trials involving 981 patients (511 receiving peripheral nerve block and 470 receiving epidural block) in the final analysis. Compared with an epidural block, a peripheral nerve block significantly reduced complications. There were no significant between-group differences in the postoperative VAS score, patient satisfaction, perioperative opioid dosage, and rehabilitation indices. Conclusions Our findings demonstrate that the peripheral nerve block is superior to the epidural block in reducing complications without compromising the analgesic effect and patient satisfaction. Therefore, a peripheral nerve block is a safe and effective postoperative analgesic method with encouraging clinical prospects.

Background: Postoperative pain management is crucial for patients undergoing total knee arthroplasty (TKA). There have been many recent clinical trials on post-TKA peripheral nerve block; however, they have reported inconsistent findings. In this meta-analysis, we aimed to comprehensively analyze studies on post-TKA analgesia to provide evidence-based clinical suggestions. Methods: We performed a computer-based query of PubMed, Embase, the Cochrane Library, and the Web of Science to retrieve related articles using neurothe following search terms: nerve block, nerve blockade, chemodenervation, chemical neurolysis, peridural block, epidural anesthesia, extradural anesthesia, total knee arthroplasty, total knee replacement, partial knee replacement, and others. After quality evaluation and data extraction, we analyzed the complications, visual analogue scale (VAS) score, patient satisfaction, perioperative opioid dosage, and rehabilitation indices. Evidence was rated using the Grading of Recommendations Assessment, Development, and Evaluation approach. Results: We included 16 randomized controlled trials involving 981 patients (511 receiving peripheral nerve block and 470 receiving epidural block) in the final analysis. Compared with an epidural block, a peripheral nerve block significantly reduced complications. There were no significant between-group differences in the postoperative VAS score, patient satisfaction, perioperative opioid dosage, and rehabilitation indices. Conclusions Our findings demonstrate that the peripheral nerve block is superior to the epidural block in reducing complications without compromising the analgesic effect and patient satisfaction. Therefore, a peripheral nerve block is a safe and effective postoperative analgesic method with encouraging clinical prospects.

Objective To evaluate the efficacy of anterior cutaneous branch of the intercostal nerve block combined with pectoral nerves (Pecs) block type Ⅱ for early postoperative analgesia by comparing with Pecs block type Ⅱ in the patients undergoing modified radical mastectomy.Methods Sixty-eight patients,aged 18-64 yr,with American Society of Anesthesiologists physical status Ⅰ-Ⅲ,scheduled for elective modified radical mastectomy under general anesthesia,were divided into 2 groups (n=34 each) using a random number table method:Pecs block type Ⅱ plus anterior cutaneous branch of intercostal nerve block group (P+A group) and Pecs block type Ⅱ group (P group).Anesthesia was induced with fentanyl,propofol and cisatracurium besilate,the patients were then tracheally intubated,and anesthesia was maintained with sevoflurane combined with nitrous oxide in both groups.In both groups,0.25％ levobupivacaine 10 ml was injected into the space between pectoralis major and pectoralis minor under ultrasound guidance,and then 0.25％ levobupivacaine 10 ml was injected into the surface of the serratus anterior muscle at the level of 3rd rib for Pecs block type Ⅱ before operation.In group P+A,0.25％ ropivacaine 10 ml (20 ml in total) was injected into the interspace between the transverse thoracic and intercostal muscles in the junction area at the level of 4th and 5th ribs to perform anterior cutaneous branch of the intercostal nerve block.The equal volume of normal saline was given instead in group P.Morphine was given for analgesia when visual analogue scale score＞3 or when the patients required.The cumulative amount of morphine administered at 24 h after surgery was recorded.The development of postoperative nausea and vomiting was observed.Results Compared with group P,the cumulative amount of morphine administered at 24 h after surgery was significantly decreased,and the postoperative analgesia time was prolonged in group P+A (P＜0.05).There was no significant difference in the incidence of postoperative nausea and vomiting between the two groups (P＞0.05).Conclusion Combination of anterior cutaneous branch of the intercostal nerve block and Pecs block type Ⅱ provides better analgesic efficacy for early postoperative analgesia than Pecs block type Ⅱ alone in the patients undergoing modified radical mastectomy.