The efficient production of L-glutamate is dependent on the product's rapid efflux, hence researchers have recently concentrated on artificially modifying its transport system and cell membrane wall structure. Considering the unique composition and structure of the cell wall of Corynebacterium glutamicum, we investigated the effects of CmpLs on L-glutamate synthesis and transport in SCgGC7, a constitutive L-glutamate efflux strain. First, the knockout strains of CmpLs were constructed, and it was confirmed that the deletion of CmpL1 and CmpL4 significantly improved the performance of L-glutamate producers. Next, temperature-sensitive L-glutamate fermentation with the CmpL1 and CmpL4 knockout strains were carried out in 5 L bioreactors, where the knockout strains showcased temperature-sensitive characteristics and enhanced capacities for L-glutamate production under high temperatures. Notably, the CmpL1 knockout strain outperformed the control strain in terms of L-glutamate production, showing production and yield increases of 69.2% and 55.3%, respectively. Finally, the intracellular and extracellular metabolites collected at the end of the fermentation process were analyzed. The modification of CmpLs greatly improved the L-glutamate excretion and metabolic flux for both L-glutamate production and transport. Additionally, the CmpL1 knockout strain showed decreased accumulation of downstream metabolites of L-glutamate and intermediate metabolites of tricarboxylic acid (TCA) cycle, which were consistent with its high L-glutamate biosynthesis capacity. In addition to offering an ideal target for improving the stability and performance of the industrial strains for L-glutamate production, the functional complementarity and redundancy of CmpLs provide a novel target and method for improving the transport of other metabolites by modification of the cell membrane and cell wall structures in C. glutamicum.
Corynebacterium glutamicum/genetics* ; Glutamic Acid/biosynthesis* ; Fermentation ; Metabolic Engineering ; Bacterial Proteins/metabolism* ; Bioreactors/microbiology* ; Gene Knockout Techniques

Zhigancao Tang （also known as Fumaitang） is a classic formula for treating "intermittent pulse and palpitations" and is widely used in clinical practice. Sanjia Fumaitang， included in the Catalogue of Ancient Classical Formulas （First Batch） published by the National Administration of Traditional Chinese Medicine of China in 2018， is derived from this formula. This paper employed bibliometric methods to comprehensively investigate and summarize the historical evolution， drug composition， herb origins and preparation， prescription meanings， and ancient and modern applications of Zhigancao Tang， analyzed the composition and usage of Zhigancao Tang， and discussed the reasons and applications of the "Fumaitang" variants created by Wu Jutong. A total of 47 valid pieces of data from 38 ancient texts were included. Results showe that Zhigancao Tang originates from the Treatise on Cold Damage （Shang Han Lun）， and the name "Fumaitang" is also recorded in the formula's description. Converted to modern measurements from the Han dynasty system， the recommended preparation for Zhigancao Tang includes 55.2 g of fried Glycyrrhizae Radix et Rhizoma， 41.4 g of Cinnamomi Ramulus， 27.6 g of Ginseng Radix et Rhizoma， 220 g of fresh Rehmannia glutinosa， 27.6 g of Asini Corii Colla， 53 g of Ophiopogonis Radix， 45 g of Cannabis Fructus， and 90 g of Jujubae Fructus. All herbs should be decocted with 1 400 mL of yellow rice wine and 1 600 mL of water until 600 mL. Once the Asini Corii Colla is fully dissolved， the decoction should be taken warm at a dosage of 200 mL， three times a day. Zhigancao Tang is effective for replenishing Qi， warming Yang， nourishing Yin， and nourishing blood and is primarily used to treat “intermittent pulse and palpitations” caused by deficiencies in heart Yin and Yang， as well as malnutrition of the heart meridian and conditions like lung atrophy. Modern applications mainly focus on cardiovascular and cerebrovascular diseases， including arrhythmias， coronary heart disease， and premature ventricular contractions. The findings from this research provide a reference for the further development of Zhigancao Tang.

Objective:To explore the current status of surgery for portal hypertension to grasp current status and future development of surgery in China.Methods:This study is jointly sponsored by China Hepatobiliary & Pancreatic Specialist Alliance & Portal Hypertension Alliance in China (CHESS).Comprehensive surveying is conducted for basic domestic situations of surgery for portal hypertension, including case load, surgical approaches, management of postoperative complications, primary effects, existing confusion and obstacles, liver transplantation(LT), laparoscopic procedures and transjugular intrahepatic portosystemic shunt(TIPS), etc.Results:A total of 8 512 cases of portal hypertension surgery are performed at 378 hospitals nationwide in 2021.Splenectomy plus devascularization predominated(53.0%)and laparoscopy accounted for 76.1%.Primary goal is preventing rebleeding(67.0%) and 72.8% of hospitals used preventive anticoagulants after conventional surgery.And 80.7% of teams believe that the formation of postoperative portal vein thrombosis is a surgical dilemma and 65.3% of hospitals practiced both laparoscopy and TIPS.The major reasons for patients with portal hypertension not receiving LT are due to a lack of qualifications for LT(69.3%)and economic factors(69.0%).Conclusions:Surgery is an integral part of management of portal hypertension in China.However, it is imperative to further standardize the grasp of surgical indications, the handling of surgical operation and the management of postoperative complications.Moreover, prospective, multi-center randomized controlled clinical studies should be performed.

OBJECTIVE To systematically evaluate the efficacy and safety of vonoprazan in the treatment of gastroesophageal reflux disease， and to provide evidence-based reference for clinical drug use. METHODS Randomized controlled trials （RCTs） about vonoprazan （trial group） versus placebo or proton pump inhibitor （control group） were searched in PubMed， the Cochrane Library， Web of Science， CNKI， Wanfang， VIP and CBM databases from the inception to June， 2022. After literature screening and data extraction， the qualities of included literature were evaluated with bias assessment tool recommended by Cochrane system evaluator manual 5.1.0. Meta-analysis， sensitivity analysis and publication bias analysis were conducted by using RevMan 5.4 software. RESULTS A total of 9 RCTs were included， involving 1 882 patients. The results of meta-analysis showed that： total response rate [OR＝1.94，95%CI（1.45，2.58），P＜0.000 01]， cure rate [OR＝2.27，95%CI（1.33，3.86），P＝0.003] and remission rate [OR＝1.81，95%CI（1.28， 2.55）， P＝0.000 7] of trial group were significantly higher than control group； there was no significant difference in the incidence of adverse drug events， diarrhea， nasopharyngitis， upper respiratory tract infection and alkaline phosphatase elevation between two groups （P＞0.05）. The results of subgroup analysis showed that cure rate of trial group was significantly higher than control group at 2 weeks of treatment （P＜0.05）； at 4 and 8 weeks of treatment， there was no significant difference in the cure rate between two groups （P＞0.05）. There was no statistically significant difference in the cure rate between two groups at 2， 4 and 8 weeks of treatment among the patients with Los Angeles grade A/B （P＞0.05）； among the patients with Los Angeles grade C/D， the cure rate of patients in the trial group was significantly higher than control group at 2， 4 and 8 weeks of treatment （P＜0.05）. The results of sensitivity analysis and publication bias analysis showed that the results of this study were robust and the possibility of publication bias was small. CONCLUSIONS Vonoprazan has a considerable effectiveness and safety in the treatment of gastroesophageal reflux disease.

OBJECTIVE To prepare cinnamaldehyde （CA） loaded liposomes bilayer-modified by bovine serum albumin （BSA）/chitosan （CTS）（BSA/CTS-Lip-CA） in order to improve the sustained-release effect and storage stability of the nanoparticles. METHODS Firstly，cinnamaldehyde loaded liposomes （Lip-CA）and blank liposomes （Lip-Blank）were prepared by thin film dispersion method. Then chitosan modified cinnamaldehyde loaded liposome （CTS-Lip-CA）and BSA/CTS-Lip-CA were obtained by electrostatic adsorption. Finally ， the prepared liposomes were characterized ， and their in vitro release characteristics and storage stability were investigated. RESULTS The particle size of BSA/CTS-Lip-CA was （177.8±4.0）nm and the Zeta potential was （－15.6±1.5）mV；they were in spherical shape ；FTIR analysis showed that the modification of BSA and CTS had no effect on the internal structure of liposomes. The results of in vitro drug release characteristics showed that the cumulative release of Lip-CA ，CTS-Lip-CA and BSA/CTS-Lip-CA within 10 hours were 82.9％，74.1％ and 72.9％ respectively. The results of storage stability showed that after 30 days of storage ，the particle sizes of Lip-CA ，CTS-Lip-CA and BSA/ CTS-Lip-CA were （134.2±2.1），（151.7±0.4），（164.8±1.5）nm；the retention rates of model drug CA were 65.4％，82.5％ and 90.2％ respectively. CONCLUSIONS BSA/CTS-Lip-CA is successfully prepared. It has a certain sustained-release effect and can improve the storage stability of the drug to a certain extent.

OBJECTIVETo evaluate the quality and clinical applicability of pyrosequencing assay kit for detecting hepatitis B virus resistance (HBV DRT).

METHODSSerial dilutions of the International Standard for HBV DNA were used to test the detection limit of the PCR for HBV DRT. Plasmids containing the either a wild-type (WT) copy or one of 10 mutant (MT) copies of the HBV RT gene were used to prepare a series of samples with various mutation ratios. To construct the linear relationship between the true mutation rate and the detected mutation rate, each sample was repeated at least 10 times. A total of 102 clinical samples were analyzed by Sanger sequencing and retested by the PCR for HBV DRT to determine the concordance of these two methods.

RESULTSThe lower detection limit of the PCR for HBV DRT was 50 IU/ml. Except for the RT236 MT, the correlation between the true mutation rate and the detected mutation rate for the other nine resistance-related mutation sites were excellent, with R² more than 0.98 (P less than 0.001). Among the 102 clinical samples, four were not amplified successfully by PCR. The results were significantly different between the PCR for HBV DRT method and the Sanger sequencing method (x² = 71.2, P less than 0.001), and concordance was observed for 897/969 (92.6%) amino acid positions in 98 samples. Concordant results were achieved in 46/98 (46.9%) samples at all 10 mutation sites. For detection of a single mutation site, concordance rates ranged from 71.5% to 100% at the 10 mutation sites, respectively. Analysis of discordant samples showed that in 87.5% (63/72), Sanger sequencing detected WT and the PCR for HBV DRT detected WT/MT. In 5.6% (4/72) of samples, Sanger sequencing detected WT/MT and the PCR for HBV DRT detected WT. In the remaining 6.9% (5/72) of samples, Sanger sequencing detected WT but PCR for HBV DRT detected MT.

CONCLUSIONThe PCR for HBV DRT showed high sensitivity and accuracy in detecting antiviral drug-resistant mutations. The method is superior to Sanger sequencing for detecting minor mutations and can be used for early detection of a resistance mutation.

Antiviral Agents ; Base Sequence ; Drug Resistance, Viral ; Hepatitis B virus ; Humans ; Mutation ; Plasmids ; Polymerase Chain Reaction ; Sequence Analysis, DNA

OBJECTIVETo evaluate the renal function in treatment-naive patients with hepatitis B virus (HBV) related cirrhosis and to identify the risk factors for renal impairment.

METHODSWe collected the data of 860 HBV-related cirrhosis patients hospitalized in our unit between Jan 1, 2011 and Dec 31, 2011. Liver function of the patients was assessed with Child-Pugh score system, and the renal function with estimated glomerular filtration rate (eGFR) calculated by Modification of Diet in Renal Disease (MDRD) equation recommended by Kidney Disease Outcomes Quality Initiative (K/DOQI). We investigated the prevalence of renal impairment (eGFR>60 ml/min/1.73 m(2)) among these patients and explored the risk factors for renal impairment.

RESULTSOf the 860 patients, 296 had complete clinical data and were included in our analysis. The overall incidence of renal impairment among the enrolled patients was 8.45% (25/296). Patients with Child-Pugh stage C showed a significantly higher incidence of renal impairment than those with stages B and A (17.17% [17/99] vs 6.67%[7/105] vs 1.09% [1/92], P<0.001). Age, history of hyperuricemia, and Child-Pugh score were identified as the risk factors for renal impairment in these patients.

CONCLUSIONIn patients with HBV-related liver cirrhosis, the incidence of renal impairment increases significantly with deterioration of the liver function, and renal function should be regularly monitored in these patients for appropriate antiviral treatment.

Adult ; Female ; Glomerular Filtration Rate ; Hepatitis B virus ; Hepatitis B, Chronic ; physiopathology ; Humans ; Incidence ; Kidney ; physiopathology ; Liver Cirrhosis ; physiopathology ; virology ; Male ; Middle Aged ; Retrospective Studies ; Risk Factors

Objective To evaluate the renal function in treatment-na?ve patients with hepatitis B virus (HBV) related cirrhosis and to identify the risk factors for renal impairment. Methods We collected the data of 860 HBV-related cirrhosis patients hospitalized in our unit between Jan 1, 2011 and Dec 31, 2011. Liver function of the patients was assessed with Child-Pugh score system, and the renal function with estimated glomerular filtration rate (eGFR) calculated by Modification of Diet in Renal Disease (MDRD) equation recommended by Kidney Disease Outcomes Quality Initiative (K/DOQI). We investigated the prevalence of renal impairment (eGFR<60 ml/min/1.73 m2) among these patients and explored the risk factors for renal impairment. Results Of the 860 patients, 296 had complete clinical data and were included in our analysis. The overall incidence of renal impairment among the enrolled patients was 8.45% (25/296). Patients with Child-Pugh stage C showed a significantly higher incidence of renal impairment than those with stages B and A (17.17%[17/99] vs 6.67%[7/105] vs 1.09%[1/92], P<0.001). Age, history of hyperuricemia, and Child-Pugh score were identified as the risk factors for renal impairment in these patients. Conclusion In patients with HBV- related liver cirrhosis, the incidence of renal impairment increases significantly with deterioration of the liver function, and renal function should be regularly monitored in these patients for appropriate antiviral treatment.

Objective To evaluate the renal function in treatment-na?ve patients with hepatitis B virus (HBV) related cirrhosis and to identify the risk factors for renal impairment. Methods We collected the data of 860 HBV-related cirrhosis patients hospitalized in our unit between Jan 1, 2011 and Dec 31, 2011. Liver function of the patients was assessed with Child-Pugh score system, and the renal function with estimated glomerular filtration rate (eGFR) calculated by Modification of Diet in Renal Disease (MDRD) equation recommended by Kidney Disease Outcomes Quality Initiative (K/DOQI). We investigated the prevalence of renal impairment (eGFR<60 ml/min/1.73 m2) among these patients and explored the risk factors for renal impairment. Results Of the 860 patients, 296 had complete clinical data and were included in our analysis. The overall incidence of renal impairment among the enrolled patients was 8.45% (25/296). Patients with Child-Pugh stage C showed a significantly higher incidence of renal impairment than those with stages B and A (17.17%[17/99] vs 6.67%[7/105] vs 1.09%[1/92], P<0.001). Age, history of hyperuricemia, and Child-Pugh score were identified as the risk factors for renal impairment in these patients. Conclusion In patients with HBV- related liver cirrhosis, the incidence of renal impairment increases significantly with deterioration of the liver function, and renal function should be regularly monitored in these patients for appropriate antiviral treatment.