ObjectiveTo observe and compare the effects of different concentrations of cyclophosphamide (CTX) in inducing premature ovarian insufficiency (POI) model in mice and investigate the mechanism of injury. MethodsThirty-two 6~8-week-old female C57BL/6J mice were randomly divided into four groups (n=8 per group) using a weight-based block randomization method. The POI model was established via a single intraperitoneal injection of 75 mg/kg cyclophosphamide (CTX), 120 mg/kg CTX, 120 mg/kg CTX + 12 mg/kg Busulfan, or an equivalent volume of normal saline (control). Ovarian coefficients, serum estradiol (E₂) and follicle-stimulating hormone (FSH) levels were measured. Western blotting was performed to assess changes in ovarian expression levels of NAD-dependent deacetylase sirtuin-5 (SIRT5) and forkhead box O3a (FOXO3a) under different modeling conditions. After determining the optimal CTX concentration for modeling, an additional forty 6~8-week-old femal C57BL/6J mice were randomly divided into five groups (n=8 per group) using a weight-based block randomization method: saline control, 120 mg/kg CTX sampling at 1, 2, 7, or 14 days after modeling. Western blotting was used to evaluate temporal changes of ovarian SIRT5 and FOXO3a protein expression. ResultsCompared with the saline control, all concentrations of CTX (75 mg/kg CTX, 120 mg/kg CTX) and 120 mg/kg CTX + 12 mg/kg Busulfan induced POI injury in mice. The 120 mg/kg CTX group exhibited smaller changes in ovarian coefficients (P<0.001) and E₂ levels (P<0.05), whereas the 120 mg/kg CTX + 12 mg/kg Busulfan group showed rough and reduced luster fur, sluggish response and was in the worst state. Compared with the saline control group, FOXO3a expression was significantly down-regulated (P<0.05), while SIRT5 remained unchanged in the 75 mg/kg CTX group (P>0.05). In contrast, both SIRT5 (P<0.05) and FOXO3a (P<0.05) were significantly down-regulated in the 120 mg/kg CTX group. Further analysis revealed that on day 2 and 7 after 120 mg/kg CTX modeling, the expressions of SIRT5 (P<0.01) and FOXO3a (P<0.001) were significantly down-regulated, with the largest decrease observed on day 7 (SIRT5, P<0.000 1; FOXO3a, P<0.000 1). ConclusionOvarian injury in the POI model induced by 120 mg/kg CTX is milder than that in the POI model induced by 75 mg/kg CTX. Moreover, the expression changes of SIRT5 and FOXO3a are most significant on day 7 after modeling induced by 120 mg/kg CTX, which may be related to the inhibition of the SIRT5-FOXO3a signaling pathway.

We have isolated an intestinal probiotic strain, Pediococcus acidilactici HRQ-1. To improve its gastrointestinal fluid tolerance, transportation and storage stability, and slow-release properties, we employed the extrusion method to prepare the microcapsules with P. acidilactici HRQ-1 as the core material and sodium alginate and chitosan as the wall material. The optimal conditions for preparing the microcapsules were determined by single factor and orthogonal tests, and the optimal ratio was determined by taking the embedding rate, survival rate, storage stability, gastrointestinal fluid tolerance, and release rate as the evaluation indexes. The results showed that under the optimal embedding conditions, the embedding rate reached (89.60±0.02)%. Under the optimal formula of freeze-drying protective agent, the freeze-drying survival rate reached (76.42±0.13)%, and the average size of the microcapsules produced was (1.16±0.03) mm. The continuous gastrointestinal fluid simulation experiments confirmed that the microcapsules ensured the viable bacterial count and can slowly release bacteria in the intestinal fluid. The curve of the viable bacterial count during storage at 4 ℃ and room temperature indicated that the prepared microcapsules achieved strains' live number protection. The formula and preparation process of P. acidilactici microcapsules may provide a technological reserve for the preparation of more live bacterial drugs in the future.
Pediococcus acidilactici/chemistry* ; Probiotics/chemistry* ; Capsules/chemistry* ; Alginates/chemistry* ; Chitosan/chemistry* ; Drug Compounding/methods* ; Glucuronic Acid/chemistry* ; Hexuronic Acids/chemistry* ; Freeze Drying

BACKGROUND:Numerous clinical studies have suggested a close relationship between obesity and osteoporosis,but whether there is a genetic causal effect between obesity and osteoporosis remains unclear. OBJECTIVE:To explore the association between obesity and osteoporosis using summary data from a large-scale genome-wide association study(GWAS)through Mendelian randomization analysis. METHODS:Obesity data were derived from summary statistics of the Genetic Investigation of Anthropometric Traits(GIANT)and the UK Biobank(UKBB).Osteoporosis data were obtained from the Genetic Factors for Osteoporosis(GeFOS)consortium,including two bone density phenotypes:total body bone mineral density(BMD)and heel BMD.The inverse variance-weighted method was the primary analysis,with the Mendelian randomization method based on Egger regression(MR-Egger)and weighted median method as supplementary approaches to calculate the causal association between genetic variations related to obesity and osteoporosis.Sensitivity analyses were conducted to validate the reliability of the results.Heterogeneity was assessed using Cochran's Q test.Horizontal pleiotropy was assessed through the MR-Egger intercept test.Leave-one-out analysis was performed to evaluate the potential influence of single nucleotide polymorphisms on the combined inverse variance-weighted estimates. RESULTS AND CONCLUSION:(1)Impact of obesity on osteoporosis:In addition to body mass index and forearm BMD,body mass index,waist-to-hip ratio,body mass index-adjusted waist-to-hip ratio,and whole-body body mass index,heel BMD,forearm BMD,lumbar spine BMD,and femoral neck BMD were causally related to each other.Further Meta-analysis revealed that obesity increased the risk of BMD(odds ratio=1.07,95%confidence interval:1.03-1.12,P<0.01).(2)Impact of osteoporosis on obesity:Apart from arm BMD and lumbar spine BMD as exposure factors showing causal relationships with obesity,other datasets indicated no causal effect between total body BMD,heel BMD,femoral neck BMD,and obesity.Additional meta-analysis demonstrated that BMD did not increase the risk of obesity(odds rate=0.99,95%confidence interval:0.98-1.01,P<0.01).There is a causal relationship between obesity and osteoporosis,suggesting that obesity may be a risk factor for osteoporosis.However,no causal association is found between osteoporosis and obesity.

Objective:To analyze and summarize the key points of design and implementation of new drug clinical trials for ankylosing spondylitis.Methods:The platform for drug clinical trial registration and information published on the official website of center for drug review and evaluation of national medical products administration (CDE) was searched to obtain data and classified statistics was conducted then. The Mean±SD and M ( Q1, Q3) were used for quantitative data for statistical description, and the rate, composition or relative ratio of qualitative data were used for statistical description. Results:A total of 23 clinical trials meeting the requirements were screened, among which 19 were biological products included in nine phase Ⅲ clinical trials. Among the four chemical drugs, two were phase Ⅱ clinical trials. One of the clinical trials on AS adopted the 1966 New York classification criteria, accounting for 4%. Nineteen of the trials adopted the1984 New York classification criteria, accounting for 83%. Three other trials adopted unspecified classification criteria, accounting for 13%. In one of these clinical trials, the age of patients included was older than 16 years old, 9 trials were 18 to 65 years old, 6 were 18 years old but without upper limit. In the definition of active AS, 19 trials took BASDAI≥4 as the cut-off value for active disease, and BASDAI, total back pain, spinal pain and morning stiffness were regarded as active disease in 4.Conclusion:The number of dosestic AS clinical trial projects continnes to rise. The 1984 classification criteria is adopted as the classification criteria in clinical trials. The minimum age in the inclusion criteria is 18 years old, there is no upper limit in age for inclusion. Disease activity can be evaluated by BASDAI score, combined with comprehensive indicators such as night-time back pain, global spinal pain and morning stiffness.

Objective To evaluate the effect of exercise-diet behavior intervention based on the transtheoretical model in patients undergoing weight loss surgery.Methods By convenience sampling,72 patients undergoing weight loss surgery in a tertiary general hospital in Xinxiang City,Henan Province from February 2021 to October 2022 were selected as the research subjects.By a random number table method,they were divided into a test group and a control group,with 36 cases in each group.The test group received exercise-diet behavior intervention based on the trans-theoretical model,while the control group received conventional intervention.The intervention began on the first day after admission and ended 6 months after surgery.The body mass index,body fat,lean body mass,diastolic blood pressure,systolic blood pressure,fasting blood glucose,insulin resistance index(HOMA-IR),Health Promoting Lifestyle Profile-Ⅱ(HPLP-Ⅱ)score,and Short-Form-36 Health Survey(SF-36)score were compared between the 2 groups before and after surgery for 3 and 6 months,as well as the complications within 6 weeks after surgery.Results The results of repeated measures analysis of variance showed that there was an interaction between the 2 groups in terms of anthropometric measurements,blood pressure and blood glucose,HPLP-Ⅱ scores,and SF-36 scores,with statistically significant differences(P＜0.001).After 6 months of surgery,the body mass index(23.32±2.32),body fat(24.10±3.46)kg,and lean body mass(41.64±3.24)kg in the test group were lower than(27.32±3.64),(28.46±4.18)kg,and(46.68±4.65)kg in the control group,and the differences were statistically significant(P＜0.001).At 3 and 6 months after operation,the diastolic blood pressure,systolic blood pressure,fasting blood glucose and HOMA-IR of the test group were lower than those of the control group,and the differences were statistically significant(P＜0.05).The HPLP-Ⅱ score of the test group was higher than that of the control group at 3 and 6 months after operation(P＜0.001).The SF-36 score of the test group was significantly higher than that of the control group at 6 months after operation(P＜0.05).The incidence of complications in the test group was 2.56％,which was not significantly different from 19.44％in the control group(P＞0.05).Conclusion The exercise-dietary behavior intervention based on the transtheoretical model can promote the formation of healthy behaviors in patients undergoing weight loss surgery,maintain weight loss effects,improve blood pressure and blood glucose levels,and enhance the quality of life of patients.

Objective To investigate the effect of inhibiting the expression of RNA component of mitochondrial RNA processing endoribonuclease(RMRP)on the proliferation and invasion of human lung cancer cell line A549 in order to provide evidence to support the research on NSCLC mechanism.Methods A total of 122 cases of patients who underwent surgical treatment were selected in Jiaozuo People's Hospital from March 2016 to March 2021.In the same period,50 healthy people from physical examination center were selected as the control group.RT-qPCR was used to detect the expressions of RMRP in blood and tissues.Human lung cancer cell line A549 was cultured and divided into si-RMRP group,siRNA-NC group and blank group.RT-qPCR,CCK-8 and Transwell assays were used to detect the expression of RMRP,proliferation activity and cell counting of invasive cells.Results The rel-ative expression level of RMRP in the blood of NSCLC patients was significantly higher than that of control group(P<0.001).The relative expression level of RMRP in NSCLC tissues was significantly higher than the adjacent tissues(P<0.001).The relative expression level of RMRP in blood and tissues of patients with poorly differenti-ated,lymph node metastasis and TNM stage Ⅲ compared with moderate to highly differentiated,no lymph node metastasis and TNM stage Ⅰ-Ⅱ were significantly increased(P<0.05).The relative expression level of RMRP in the cells of the si-RMRP group was lower than that of blank and the siRNA-NC group(P<0.001).Compared with the blank group and the siRNA-NC group,the absorbance(A)value of cells at 24,48,72 and 96 h in the si-RMRP group was decreased(P<0.05).The number of invasion cells in the si-RMRP group was lower than that in the blank group and the siRNA-NC group(F=27.765,P<0.001).Conclusions The relative expression levels of RMRP in blood and tissues of NSCLC patients are increased.Down-regulation of the expres-sion of RMRP gene in A549 cells can inhibit cell proliferation and reduce cell invasion.

Objective　To investigate the effect of laboratory domestication duration on the gut microbiota of mosquitoes, reveal the patterns of intestinal microbiota changes, and more systematically elucidate the role of mosquito gut microbiota on the transmission of vector-borne pathogens and mosquito resistance to insecticides, providing new insights into the interaction between microorganisms and mosquitoes. Furthermore, to understand the composition of gut microbiota in different geographic strains of mosquitoes to serve as a reference for selecting suitable populations for subsequent functional studies of gut microbiota. Methods The full-length bacterial ribosomal 16S amplification and metagenomic sequencing were performed to reveal the midgut microbiota in 5 Aedes aegypti strains which had been domesticated in the laboratory for various years. The composition, abundance, diversity, and correlation of the microbiota carried by the 5 mosquito strains were analyzed through bioinformatics, and the content of specific bacteria that play an important role in mosquito growth and development, pathogen transmission, and insecticide resistance in different samples were compared. Results The ML and CJ strains, which had been domesticated for a shorter duration, exhibited higher diversity indices compared to the DB and SA strains, which had been bred for a longer time. The gut microbiota of all mosquitoes mainly consisted of Proteobacteria, Firmicutes, and Bacteroidota. At the genus level, the bacterial composition of Aedes aegypti MH, DB, and SA strains was relatively homogenous, predominantly consisting of Cedecea and Asaia, while the ML and CJ strains, domesticated for a shorter period, harbored a richer and more balanced variety of bacteria. Conclusions Under laboratory conditions, the diversity of mosquito gut microbiota will decrease with increased domestication duration. Bacteria that have a positive effect on mosquito growth and development tend to be retained and increase in proportion over time, while the proportion of most other bacteria will decrease or even disappear.

Objective:To compare the efficacy and safety of different dosages of new drugs in the treatment of PsA by using network meta-analysis.Methods:Three medical databases (PubMed, Web of Science, Cochrane Library) were searched for the studies that compared the efficacy and safety of 4 new drugs (secukinumab, ixekizumab, apremilast, tofacitinib) with different dosages in the treatment of PsA. Data from included studies were analyzed by Stata 15.0.Results:A total of 16 RCTs were included. The results of the network meta-analysis showed that: (1) Among the overall patients, in terms of ACR20 response rate, the larger the surface under the cumulative ranking (SUCRA), the more effective it is. Secukinumab 300 mg Q4W(96.1%) had the best efficacy, followed by ixekizumab 80 mg Q4W(79.0%), ixekizumab 80 mg Q2W(75.1%), secukinumab 150 mg Q4W(73.2%), apremilast 30 mg BID(50.6%), apremilast 20 mg BID(38.6%), tofacitinib 5 mg BID(18.1%), tofacitinib 10 mg BID(17.7%) and placebo(2.0%). (2) In terms of PASI75 response rate, the larger the area under the SUCRA curve, the more effective it is. Ixekizumab 80 mg Q4W(96.1%) had the best efficacy, followed by ixekizumab 80 mg Q2W(88.7%), secukinumab 300 mg Q4W(75.6%), secukinumab 150 mg Q4W(63.3%), apremilast 30 mg BID(44.5%), apremilast 20 mg BID(38.4%), tofacitinib 10 mg BID(30.0%), tofacitinib 5 mg BID(12.5%) and placebo(1.0%). (3) Among the overall patients, in terms of safety, the smaller the area under the SUCRA curve, the higher the safety it is. Secukinumab 300 mg Q4W (17.3%) has the best safety. (4) The results of subgroup analysis showed that in terms of ACR20 response rate, ixekizumab 80 mg Q2W(85.3%) had the best efficacy in bDMARDs-na?ve patients, while in bDMARDs-IR patients, secukinumab 300 mg Q4W(83.9%) had the best efficacy.Conclusion:Among all patients, secukinumab 300 mg Q4W is the best in terms of ACR20 response rate and safety, but ixekizumab 80 mg Q4W is more effective in improving PsA lesions comparing yo other drugs.

Objective:To investigate the clinical effect of CAG stimulating regimen for refractory adult early T cell precursor acute lymphoblastic leukemia (ETP-ALL) complicated with fusarium infection and the clinical features as well as antifungal strategy of cutaneous fusarium infection.Methods:The diagnosis and treatment of 1 adult patient diagnosed as ETP-ALL complicated with cutaneous fusarium infection in the First Hospital of Jilin University in September 2020 were retrospectively analyzed, and related literatures were reviewed.Results:VICP chemotherapy regimen showed no effectiveness in this patient who was presented with persistent agranulocytosis complicated with cutaneous fusariosis infection. After amphotericin B therapy for infection, he achieved the stable disease and successfully underwent CAG stimulating regimen salvage treatment. The minimal residual disease turned into negative after consolidation chemotherapy based on the myeloid regimen. Finally this patient survived from haploid allogeneic hematopoietic stem cell transplantation after consolidation chemotherapy and fusarium was under the control by using posaconazole as secondary prevention therapy.Conclusions:CAG stimulating regimen can be recommended as reinduction therapy for relapsed/refractory ETP-ALL. Sequential therapy of amphotericin B followed by posaconazole can be a useful antifungal strategy for fusarium infection.