Objective:To investigate variation of peripheral blood monocyte subtypes in patients with rheumatoid arthritis(RA),to study variation of surface antigen expression intensity and plasma related cytokine content of monocytes of each subtype,and to discuss their clinical application value.Methods:Percentage of monocyte subtypes and mean fluorescence intensity(MFI)of monocyte surface markers(HLA-DR,CD64,CD11b)in peripheral blood of 38 patients with RA were detected by multi-parameter flow cytometry,and differences were statistically analyzed compared with 40 healthy controls(HV group).Plasma levels of five cyto-kines(IL-2,IL-6,IL-10,TNF-α and IFN-γ)were detected by flow fluorescence microsphere technology.Linear correlation analysis was performed to determine correlation between percentage of monocyte subtypes and MFI of their surface markers and expression of related cytokines in RA patients.Disease activity 28(DAS28)score was performed in RA patients to study its association with percent-age of monocyte subtypes and plasma cytokines.Results:Compared with control group,percentage of classical monocytes in RA pa-tients decreased significantly(P<0.05),while percentage of intermediate monocytes and non-classical monocytes increased signifi-cantly(P<0.01).HLA-DR expression(MFI)on monocyte subsets in RA group was significantly higher than HV group(P<0.05 or P<0.01).CD64 expression(MFI)on intermediate monocytes and non-classical monocytes in RA group was significantly higher than HV group(P<0.01 or P<0.001).CD11b expression(MFI)on intermediate monocytes in RA group was significantly higher than HV group(P<0.001).In RA group,percentage of intermediate monocytes was positively correlated with contents of IL-6,TNF-α and IFN-γ(P<0.01).HLA-DR expression(MFI)on intermediate monocytes was positively correlated with TNF-α content(P<0.05);CD64 expres-sion(MFI)on non-classical monocytes was positively correlated with contents of IL-2 and IL-10(P<0.05).In RA group,percentage of intermediate monocytes was positively correlated with DAS28(P<0.001);plasma IL-6 and TNF-α levels were positively correlated with DAS28(P<0.01).Conclusion:Characteristic expression of intermediate monocytes and their surface antigens are closely related to occurrence of RA,and related inflammation is triggered by secretion of related cytokines.Monocyte subtype detection can be a new experimental diagnostic index for RA.Dynamic monitoring of peripheral blood monocyte subtypes and plasma IL-6 and TNF-α in RA patients during treatment is helpful to guide clinical treatment of RA.

BACKGROUND:Percutaneous kyphoplasty through a unilateral puncture approach is considered more effective than a bilateral puncture approach for the treatment of osteoporotic vertebral compression fractures.However,due to the unique anatomical shape and location of the L5 vertebral body,the traditional unilateral puncture approach has limited applicability in treating L5 osteoporotic vertebral compression fractures.To address this issue,we propose the use of the"O"point puncture approach.OBJECTIVE:To compare the effectiveness of percutaneous kyphoplasty via basal transversal process-posterior superior pedicle-lateral articular process intersection("O"point)with conventional approach percutaneous kyphoplasty in treating the L5 osteoporotic vertebral compression fractures.METHODS:A retrospective analysis was conducted on 54 patients with L5 osteoporotic vertebral compression fractures treated with traditional percutaneous kyphoplasty or percutaneous kyphoplasty via the"O"point approach between January 2020 and December 2022 at Affiliated Hospital of Southwest Medical University.According to the surgical method,the patients were divided into the"O"point approach group(group A,n=29)and the traditional approach group(group B,n=25).The position of the"O"point and iliac spine and puncture angle were measured by orthography before operation in group A.Cobb angle,anterior vertebral margin,middle height,and bone cement distribution were compared between the two groups prior to and 2 days after surgery,and during final follow-up.Pain relief and daily living ability were assessed using the visual analog scale and Oswestry disability index prior to and 2 days after surgery,and during final follow-up.Complications related to the surgery were recorded.RESULTS AND CONCLUSION:(1)Operative time and intraoperative fluoroscopy times were significantly shorter in group A than in group B(P＜0.05).However,no significant differences were observed in intraoperative blood loss or bone cement injection amounts between the two groups.(2)Both groups demonstrated significantly lower visual analog scale scores and Oswestry disability index on day 2 after surgery and at the last follow-up in comparison to before surgery.However,group A experienced a more noticeable decrease than group B(P＜0.05).(3)On both sides,the"O"point was located on average 1.23 cm below the highest point of the iliac spine line.The mean transverse distance between the"O"point and the iliac spine was 2.89 cm.(4)During the final follow-up,the Cobb angle of both groups exhibited significant improvements compared to their respective preoperative values(P＜0.05).However,there was no significant difference between the two groups(P＞0.05).(5)After operation,the distribution of bone cement was good in group A,accounting for 97％(28/29),and good in group B,accounting for 88％(22/25).Group A was significantly better than group B(P＜0.05).(6)The efficacy of percutaneous kyphoplasty utilizing the"O"point puncture technique on osteoporotic vertebral compression fractures of the L5 vertebra was superior to that of the traditional percutaneous kyphoplasty approach.However,the bone cement distribution achieved with percutaneous kyphoplasty via"O"point puncture was more satisfactory,and the puncture approach was able to circumvent the influence of a high iliac spine on the L5 vertebra.

O-acetyl-L-homoserine (OAH) is a promising platform compound for the production of L-methionine and other valuable compounds, while its low yield and low conversion rate limit the industrial application. To solve these problems, we constructed a strain for high OAH production with the previously constructed L-homoserine producer Escherichia coli HS33 as the chassis by systematic metabolic engineering. Firstly, PEP accumulation, pyruvate utilization, and OAH synthesis pathway (overexpressing aspB, aspA, and thrA^C1034T) were enhanced to obtain an initial strain accumulating 13.37 g/L OAH. Subsequently, the co-factor synthesis genes were integrated to supply reducing power and energy, which increased the yield to 15.79 g/L. The OAH yield of the engineered strain OAH28 was further increased to 17.49 g/L by strengthening the acetic acid reuse pathway, improving the supply of acetyl-CoA, and regulating the expression of MetX from different sources. Finally, in a 5 L fermenter, OAH28 achieved an OAH titer of 47.12 g/L, with a glucose conversion rate of 32% and productivity of 0.59 g/(L·h). The results lay a foundation for increasing the OAH production by metabolic engineering and give insights into the industrial production of OAH.
Metabolic Engineering/methods* ; Escherichia coli/genetics* ; Homoserine/biosynthesis* ; Fermentation

Echinocandin B (ECB) is a key precursor of the antifungal drug anidulafungin. It is a secondary metabolite of Aspergillus nidulans, and its titer in fermentation is significantly affected by the ECB synthesis pathway and cell morphology. In this study, the key genes related to the transcription activation, hydroxylation, and cell morphology during ECB biosynthesis were investigated to increase the fermentation titer of ECB and to change the cell morphology of Aspergillus nidulans to reduce the viscosity of the fermentation broth. The results indicated that after overexpression of ecdB and ecdK, the ECB titer increased by 25.8% and 23.7%, respectively, compared with that of the wild-type strain, reaching (2 030.5±99.2) mg/L and (1 996.4±151.4) mg/L. However, the deletion of fksA associated with cell wall synthesis resulted in damage to the cell wall, affecting strain growth and product synthesis. The engineered strain overexpressing ecdB was fermented in a 50-L bioreactor, in which the ECB titer reached 2 234.5 mg/L. The findings laid a research foundation for the subsequent metabolic engineering of this strain.
Fermentation ; Aspergillus nidulans/genetics* ; Echinocandins/genetics* ; Bioreactors/microbiology* ; Fungal Proteins/biosynthesis* ; Metabolic Engineering

Abstract In recent years, the prevalence of overweight and obesity among children and adolescents has risen rapidly, posing a serious threat to their physical and mental health. To provide scientific, systematic, and standardized weight management guidance for overweight and obese children and adolescents, the study focuses on the core concept of healthy lifestyle intervention, integrates multidisciplinary expert opinions and research findings,and proposes a comprehensive multidisciplinary intervention framework covering scientific exercise intervention, precise nutrition and diet, optimized sleep management, and standardized psychological support. It calls for the establishment of a multi agent collaborative management mechanism led by the government, implemented by families, fostered by schools, initiated by individuals, optimized by communities, reinforced by healthcare, and coordinated by multiple stakeholders. Emphasizing a child and adolescent centered approach, the consensus advocates for comprehensive, multi level, and personalized guidance strategies to promote the internalization and maintenance of a healthy lifestyle. It serves as a reference and provides recommendations for the effective prevention and control of overweight and obesity, and enhancing the health level of children and adolescents.

This study aims to evaluate the effects of sodium butyrate(NaBu)on preliminary follicle development of mice,using an in vitro 3D culture system.Preantral follicles were isolated from mouse ovaries(12.5 days post-birth)and cultured in vitro,NaBu was added for treatment during the culture process.Follicle morphology was observed and follicular diameter,survival rate,antral formation rate,oocyte maturation rate and early embryonic development were evaluated.The re-sults showed that an in vitro 3D culture system for preantral follicles of mice was successfully es-tablished.Matured follicles were cultured following an initial induction period of 8 days,after which M Ⅱ-stage oocytes were effectively expelled.During the process of follicle culture,the addition of varying doses(0.05-1.00 mmol/L)of NaBu from 0th day to 8 days had not showed any significant promotion in follicle development at any of the tested concentrations.NaBu at a dose of 0.10 mmol/L significantly increased antral formation rate(treated on 4th day for 2 days).In conclusion,short-term administration of appropriate doses of NaBu could significantly facilitate in vitro preantral follicle development and oocyte maturation by ensuring sustained developmental potential within follicles.This study may provide a theoretical basis for sodium butyrate to improve the reproduc-tive capacity of livestock and poultry in the breeding process.

The aim of this experiment was to study the effects of different feeding modes on growth performance,blood biochemical indexes and intestinal flora of lactating Holstein male calves.Twenty-four newborn Holstein male calves with body mass of(40.00±1.01)kg and similar day old were selected and randomly divided into four groups of six calves each.The subgroups were low-milk group(LM),high-milk group(HM),high-milk milk replacer feeding group(HMR),and low-milk switching to high-milk milk replacer feeding group(CMR).The results showed that:At 45 d,the body mass of calves in the HM group was significantly higher than that of calves in the other groups(P＜0.05),and at 60 d,the body mass of calves in the HM group was significantly higher than that of calves in the LM &.CMR groups(P＜0.05).At 90 d,the body mass of calves in the LM group was significantly higher than that of calves in the HM group.Throughout the ex-perimental period,the average daily weight gain and average pellet feed intake of calves in the LM group were significantly higher than that of calves in the HM group(P＜0.05).The calf globulin level in the HMR group was significantly higher than that in the LM and HM groups(P＜0.05);the plasma immunoglobulin A level of calves in the HM group was significantly lower than that of calves in the LM and HMR groups(P＜0.05);and the plasma immunoglobulin M level of calves in the HM group was significantly higher than that of calves in the LM and CMR groups(P＜0.05),and HMR group was also significantly higher than that of LM group(P＜0.05);plasma glutathione peroxidase level of calves in HMR group was significantly higher than that of LM group(P＜0.05);plasma malondialdehyde level of calves in LM group was significantly higher than that of calves in HMR and HM groups(P＜0.05),and CMR group was also significantly higher than that of HM group(P＜0.05).Relative abundance of Thermodesulfovibrio was higher in the HM group(P＜0.05),relative abundance of Bacteroidetes in the LM group was significantly higher than that in the HMR and HM groups(P＜0.05),relative abundance of Blautia in the HM group(P＜0.05),and relative abundance of Corynebacterium in the CMR group was significantly higher than that in the LM and HM groups(P＜0.05).In summary,calves in the LM group had better weaning weights and pellet feed intake;calves in the CMR group could compensate for growth by supplemental feeding of milk replacer to obtain more optimal weaning weights and pel-let feed intake;the HMR group proved that milk-free feeding could ensure stable growth of calves;and calves in the HM group had a better pre-lactation growth performance,lower levels of oxida-tive stress,and a healthier fecal flora.

This study aims to evaluate the effects of sodium butyrate(NaBu)on preliminary follicle development of mice,using an in vitro 3D culture system.Preantral follicles were isolated from mouse ovaries(12.5 days post-birth)and cultured in vitro,NaBu was added for treatment during the culture process.Follicle morphology was observed and follicular diameter,survival rate,antral formation rate,oocyte maturation rate and early embryonic development were evaluated.The re-sults showed that an in vitro 3D culture system for preantral follicles of mice was successfully es-tablished.Matured follicles were cultured following an initial induction period of 8 days,after which M Ⅱ-stage oocytes were effectively expelled.During the process of follicle culture,the addition of varying doses(0.05-1.00 mmol/L)of NaBu from 0th day to 8 days had not showed any significant promotion in follicle development at any of the tested concentrations.NaBu at a dose of 0.10 mmol/L significantly increased antral formation rate(treated on 4th day for 2 days).In conclusion,short-term administration of appropriate doses of NaBu could significantly facilitate in vitro preantral follicle development and oocyte maturation by ensuring sustained developmental potential within follicles.This study may provide a theoretical basis for sodium butyrate to improve the reproduc-tive capacity of livestock and poultry in the breeding process.

Objective:To investigate variation of peripheral blood monocyte subtypes in patients with rheumatoid arthritis(RA),to study variation of surface antigen expression intensity and plasma related cytokine content of monocytes of each subtype,and to discuss their clinical application value.Methods:Percentage of monocyte subtypes and mean fluorescence intensity(MFI)of monocyte surface markers(HLA-DR,CD64,CD11b)in peripheral blood of 38 patients with RA were detected by multi-parameter flow cytometry,and differences were statistically analyzed compared with 40 healthy controls(HV group).Plasma levels of five cyto-kines(IL-2,IL-6,IL-10,TNF-α and IFN-γ)were detected by flow fluorescence microsphere technology.Linear correlation analysis was performed to determine correlation between percentage of monocyte subtypes and MFI of their surface markers and expression of related cytokines in RA patients.Disease activity 28(DAS28)score was performed in RA patients to study its association with percent-age of monocyte subtypes and plasma cytokines.Results:Compared with control group,percentage of classical monocytes in RA pa-tients decreased significantly(P<0.05),while percentage of intermediate monocytes and non-classical monocytes increased signifi-cantly(P<0.01).HLA-DR expression(MFI)on monocyte subsets in RA group was significantly higher than HV group(P<0.05 or P<0.01).CD64 expression(MFI)on intermediate monocytes and non-classical monocytes in RA group was significantly higher than HV group(P<0.01 or P<0.001).CD11b expression(MFI)on intermediate monocytes in RA group was significantly higher than HV group(P<0.001).In RA group,percentage of intermediate monocytes was positively correlated with contents of IL-6,TNF-α and IFN-γ(P<0.01).HLA-DR expression(MFI)on intermediate monocytes was positively correlated with TNF-α content(P<0.05);CD64 expres-sion(MFI)on non-classical monocytes was positively correlated with contents of IL-2 and IL-10(P<0.05).In RA group,percentage of intermediate monocytes was positively correlated with DAS28(P<0.001);plasma IL-6 and TNF-α levels were positively correlated with DAS28(P<0.01).Conclusion:Characteristic expression of intermediate monocytes and their surface antigens are closely related to occurrence of RA,and related inflammation is triggered by secretion of related cytokines.Monocyte subtype detection can be a new experimental diagnostic index for RA.Dynamic monitoring of peripheral blood monocyte subtypes and plasma IL-6 and TNF-α in RA patients during treatment is helpful to guide clinical treatment of RA.

BACKGROUND:Percutaneous kyphoplasty through a unilateral puncture approach is considered more effective than a bilateral puncture approach for the treatment of osteoporotic vertebral compression fractures.However,due to the unique anatomical shape and location of the L5 vertebral body,the traditional unilateral puncture approach has limited applicability in treating L5 osteoporotic vertebral compression fractures.To address this issue,we propose the use of the"O"point puncture approach.OBJECTIVE:To compare the effectiveness of percutaneous kyphoplasty via basal transversal process-posterior superior pedicle-lateral articular process intersection("O"point)with conventional approach percutaneous kyphoplasty in treating the L5 osteoporotic vertebral compression fractures.METHODS:A retrospective analysis was conducted on 54 patients with L5 osteoporotic vertebral compression fractures treated with traditional percutaneous kyphoplasty or percutaneous kyphoplasty via the"O"point approach between January 2020 and December 2022 at Affiliated Hospital of Southwest Medical University.According to the surgical method,the patients were divided into the"O"point approach group(group A,n=29)and the traditional approach group(group B,n=25).The position of the"O"point and iliac spine and puncture angle were measured by orthography before operation in group A.Cobb angle,anterior vertebral margin,middle height,and bone cement distribution were compared between the two groups prior to and 2 days after surgery,and during final follow-up.Pain relief and daily living ability were assessed using the visual analog scale and Oswestry disability index prior to and 2 days after surgery,and during final follow-up.Complications related to the surgery were recorded.RESULTS AND CONCLUSION:(1)Operative time and intraoperative fluoroscopy times were significantly shorter in group A than in group B(P＜0.05).However,no significant differences were observed in intraoperative blood loss or bone cement injection amounts between the two groups.(2)Both groups demonstrated significantly lower visual analog scale scores and Oswestry disability index on day 2 after surgery and at the last follow-up in comparison to before surgery.However,group A experienced a more noticeable decrease than group B(P＜0.05).(3)On both sides,the"O"point was located on average 1.23 cm below the highest point of the iliac spine line.The mean transverse distance between the"O"point and the iliac spine was 2.89 cm.(4)During the final follow-up,the Cobb angle of both groups exhibited significant improvements compared to their respective preoperative values(P＜0.05).However,there was no significant difference between the two groups(P＞0.05).(5)After operation,the distribution of bone cement was good in group A,accounting for 97％(28/29),and good in group B,accounting for 88％(22/25).Group A was significantly better than group B(P＜0.05).(6)The efficacy of percutaneous kyphoplasty utilizing the"O"point puncture technique on osteoporotic vertebral compression fractures of the L5 vertebra was superior to that of the traditional percutaneous kyphoplasty approach.However,the bone cement distribution achieved with percutaneous kyphoplasty via"O"point puncture was more satisfactory,and the puncture approach was able to circumvent the influence of a high iliac spine on the L5 vertebra.