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

【Objective】 To investigate the protective effects of aflexible sleeve penile protection device on reducing postoperative pain and wound edema in patients after circumcision. 【Methods】 A total of 54 patients who underwent circumcision at Yan’an Branch of Peking University Third Hospital during Feb.1 and May 31, 2023 were enrolled.The patients were randomly divided into the experimental group and control group, with 27 patients in either groups.Patients in the experimental group were treated with a flexible sleeve penis protection device after surgery, and patients in the control group were treated with traditional gauze bandage after surgery.Postoperative pain, wound edema and complications were compared between the two groups. 【Results】 In terms of pain, the visual analogue scale of the experimental group was significantly lower at 6 hours [(1.7±0.9) vs.(3.3±1.9), P<0.001] and 2 days [(2.0±1.3) vs.(3.3±1.3), P<0.001] after surgery than that of the control group, but there were no statistically significant differences between the two groups on the 4th and 7th postoperative days (P>0.05).In terms of edema, the edema score of the experimental group was significantly lower than that of the control group on the 2nd postoperative day [(2.0±1.0) vs.(4.0±0.8), P<0.001] , the 4th postoperative day [(1.5±1.2) vs.(2.6±0.9), P<0.001] , and the 7th postoperative day [(0.9±1.3) vs.(2.3±1.5), P<0.001] .There was no statistically significant difference in the incidence of complications between the two groups (P>0.05). 【Conclusion】 The flexible sleeve penile protection device has significant effects of reducing early postoperative pain and reducing edema in patients undergoing circumcision.

Aim To investigate the mechanism and search for potential biomarkers of ovalbumin ( OVA ) -induced asthma in mice base on lipidomics. Methods A BALB/c mouse model of asthma was prepared by OVA. TNF-α, IL-4, IL-10, IFN-γ levels in BALF and IgE level in serum were measured by ELISA. The inflammatory changes in mouse lung tissue were observed using HE staining. Lipid mediators ( LMs) in lung tissue and serum were quantified with UPLC-MS/ MS strategy. Results IgE level in serum and TNF-α, IFN-γ levels in BALF were higher (P <0.05) of asthmatic mice.Typical inflammatory manifestations were seen in lung tissue of asthmatic mice. A total of 57 lipid mediators were quantified with UPLC-MRM. LMs metabolic profiles differed significantly in serum and lung tissue between asthmatic and normal mice, 17 significantly different LMs were found in lung tissue and 6 LMs were found in serum, and the differential metabolites were produced through the cyclooxygenase (COX), lipoxygenase (LOX) and cytochrome P450 oxidase (P450) metabolic pathways. Conclusions OVA-induced allergic asthma can cause disorder of lip-id mediators, LMs and cytokines are involved in the occurrence and development of asthma. The differential LMs have potential research value as biomarkers for the development of allergic asthma.

Prostate cancer is the most common male malignant tumor in the world, and its death toll is second only to lung cancer. Androgen deprivation therapy (ADT) is a major treatment for prostate cancer besides radical surgery. ADT treatment will lead to the inevitable progression of prostate cancer patients to castration-resistant prostate cancer (CRPC). The current research results have confirmed that the transformation from androgen deprivation prostate cancer (ADPC) to CRP is related to the reactivation of the androgen receptor signal pathway. In this review, the research progress on the mechanism of the androgen receptor signaling pathway in CRPC was reviewed in order to provide a scientific basis and new ideas for the diagnosis and treatment of CRP.

Objective: To investigate the mutation of P4HA2 gene in Tujia high myopia patients. Methods: Clinical data and genomic DNA were collected from 288 Tujia patients with high myopia, whose spherical error ≥-6.00 diopters and axial length≥26 mm.All coding exons regions of P4HA2 were screened in patients to detect causative mutation by Sanger sequencing.The detected mutation was further screened in 192 normal control chromosomes in the same district.The pathogenicity of genetic mutations was predicted through bioinformatics analysis.This study followed the Declaration of Helsinki.All patients or their guardians signed informed consent. Results: Four variations of P4HA2 gene were found in 288 patients, including one missense mutations(c.145C>A), two in-containing mutations (c.1306-62C>T, c.82+ 22C>T) and one insertion mutation (c.179+ 16_179+ 17 ins T). Missense mutation c. 145C>A was predicted as suspicious pathogenic gene by Polyphen2.According to the standard of ACMG in the United States, the variation was uncertain in pathogenicity. Conclusions Missense mutation c. 145C>A in P4HA2 gene is a suspicious pathogenic gene mutation in Tujia patients with high myopia.

OBJECTIVE: To investigate the effect of calenduloside E on lipopolysaccharide (LPS)-induced inflammatory response in RAW264.7 cells and explore the underlying molecular mechanism. METHODS: CCK-8 assay was used to examine the effect of different concentrations of calenduloside E (0-30 μg/mL) on the viability of RAW264.7 cells. The release of the pro-inflammatory cytokines tumor necrosis factor-α (TNF-α) and interleukin-1β (IL-1β) in RAW264.7 cells in response to pretreatment with 6, 8, and 10 μg/mL calenduloside E for 2 h followed by stimulation with 100 ng/mL LPS was detected using enzyme-linked immunosorbent assay (ELISA). The expression levels of iNOS and COX-2 and the activation of JAK-stats, MAPKs and NF-кB signaling pathways in the treated cells were determined using Western blotting. A reactive oxygen species (ROS) detection kit was used to detect ROS production in the cells, and the nuclear translocation of the transcription factor stat3 was observed by laser confocal microscopy. RESULTS: Calenduloside E below 20 μg/mL did not significantly affect the viability of RAW264.7 cells. Calenduloside E dose-dependently decreased the expression levels of iNOS and COX-2 induced by LPS, inhibited LPS-induced release of TNF-α and IL-1β, and suppressed LPS-induced JAK1-stat3 signaling pathway activation and stat3 nuclear translocation. Calenduloside E also significantly reduced ROS production induced by LPS in RAW264.7 cells. CONCLUSIONS Calenduloside E inhibits LPS-induced inflammatory response by blocking ROS-mediated activation of JAK1-stat3 signaling pathway in RAW264.7 cells.
Animals ; Lipopolysaccharides ; Mice ; NF-kappa B ; Oleanolic Acid ; analogs & derivatives ; RAW 264.7 Cells ; Reactive Oxygen Species ; Saponins ; Signal Transduction

Objective: To analyze the influence of the confidence to pre-exposure prophylaxis (PrEP) on the willingness to use PrEP in men who have sex with men (MSM) by using the path analysis model based on structural equation model approach. Methods: A total of 550 MSM were recruited in Urumqi by snowball method and a questionnaire survey was conducted among them. According to the professional knowledge and analysis results of the confidence and willingness to use PrEP, structural equation model (SEM) analysis method was used to construct a path analysis model. Results: A total of 513 MSM participated in the survey. The modified path equation model was well fitted, with the modified fitting index as: GFI=0.993, RMSEA<0.001, and AGFI=0.984. The confidence to PrEP had direct influence on the willingness; the degree of influence from sex partners, the attitude of sex partner to PrEP and the positive emotions not only had direct effects on willingness of PrEP use, but also had indirect effects on willingness of PrEP use by affecting the confidence to it; the role in sexual behavior, AIDS severity, HIV prevention behavior had direct effects on willingness of PrEP use. The proportion of HIV infection in the population had no direct effects on willingness of PrEP use, but had indirect effects on willingness of PrEP use by affecting the confidence to it. Conclusions The confidence to PrEP had influence on willingness of PrEP use in MSM, therefore targeted activities can be conducted to improve the confidence and willingness of MSM in taking the PrEP and reducing the risk of HIV infection in MSM. Compared with the traditional multiple regression analysis, the path analysis using the structural equation model could better reveal the mediating effect between the independent variables and dependent variables.

OBJECTIVETo study the antalgic and antiphlogistic functions and mechanism of ronggudingtong (RGDT) plaster (traditional Chinese medicine).

METHODSThe painful models were established with hot plate test or acetic acid writhing and the inflammatory models were established with daubing dimethylbenzene on auricle or injecting formaldehyde in toe or synovial envelope to study the antalgic and antiphlogistic functions of RGDT Plaster. The total protein and leukotriene B4(LTB4) in inflammatory exudate were detected to investigate the antalgic and antiphlogistic mechanism of RGDT plaster. The mice were randomly divided into different groups (n = 11), on the basis of drug using, the indexes of pain threshold, swelling degree were observed. Sixty-six mice were used to establish gasbag synovitis model and randomly divided into normal control group,model control group, positive control group (Voltaren gel 0.8 mg/d)and low/medium/high dosage RGDT plaster treating groups(30 mg/d, 60 mg/d, 120 mg/d).

RESULTS30 mg/d, 60 mg/d,120 mg/d RGDT plaster could upgrade the pain thresholds, remit auricular and foot swelling (P < 0.05, P < 0.01), and degrade total protein and LTB4 in inflammatory exudates (P < 0.05, P < 0.01).

CONCLUSIONRGDT plaster has some antalgic and antiphlogistic functions, and one of the mechanisms is depressing synthesis of LTB4.

Analgesics ; pharmacology ; Animals ; Anti-Inflammatory Agents ; pharmacology ; Leukotriene B4 ; metabolism ; Medicine, Chinese Traditional ; Mice ; Pain ; drug therapy

Objective Human bone marrow mesenchymal stem cells were transfected by Smad7 and labeled with green fluores-cent mark.BMMSCs were implanted into rabbit glaucoma operational model and observed surviving condition.Methods Through BP and LR reaction,Smad7 with green fluorescent mark was inserted into human bone marrow mesenchymal stem cells by bacterio-phage,filtered positive colony and picked out cell line.15 New Zealand white rabbits were enforced trabeculectomy with BMMSC, then following up cell survival condition.Results Smad7 expressed stable in human bone marrow mesenchymal stem cells with sat-isfactory green fluorescent mark.BMMSC survived in rabbit trabecula with stable green fluorescent and effective ocular press.Con-clusion Smad7 with green fluorescent mark could be inserted into human bone marrow mesenchymal stem cells stably,and has ef-fective results in rabbit model.

OBJECTIVETo study the application of VSD in the treatment of severe necrotizing fasciitis in extremities of patients.

METHODSEight patients, suffering from severe necrotizing fasciitis, who had been traditionally treated with iodophor-soaked gauze for 21 to 365 days in other hospitals, were transferred to our institute because of the nonhealing wounds and systemic toxic symptoms induced by infection, from January 2011 to August 2013. After admission, surgical debridement was performed timely, and the necrotic tissue was collected during the operation for pathological observation after HE staining. After the operation, VSD was started with negative pressure ranging from -100 to -80 kPa, and the furacilin solution (0.2 g/L) and oxygen (2 L/min) were continuously infused into the wound during the treatment. Surgical debridement was performed repeatedly according to the wound condition followed by change of VSD dressings to continue VSD treatment. The wounds were closed by suturing or with autologous skin grafts after being covered by fresh granulation tissue. The times of surgical debridement, times of change of VSD materials, wound healing status, and length of stay in our institute were recorded. All patients were followed up for a long time. Results HE staining showed that there were diffuse necrotic adipose and fibrous connective tissues in the necrotic tissue, and the normal tissue structure disappeared accompanied by significant infiltration of inflammatory cells. The number of surgical debridement was 2 to 10 (3.9 +/- 2.8) times. The number of VSD materials change was 2 to 10 (4.0 +/- 2.9) times. Wounds were closed by suturing and healed in two patients; wounds in the other six patients were partially sutured, their residual wounds were healed by autologous skin grafting. The length of stay in our institute was 20 to 49 (33 +/- 10) days. All patients were discharged after recovery. Patients were followed up for 2 to 24 months, and their wounds were found to be in good condition without ulceration or recurrence.

CONCLUSIONSVSD can effectively remove the necrotic tissues and exudates from the fascial spaces and promote proliferation of granulation tissue. Therefore it serves as an effective approach to the treatment of severe necrotizing fasciitis in extremities.

Debridement ; Drainage ; Extremities ; surgery ; Fasciitis, Necrotizing ; surgery ; Granulation Tissue ; Humans ; Negative-Pressure Wound Therapy ; Oxygen ; Pressure ; Skin ; Skin Transplantation ; Ulcer ; Vacuum