OBJECTIVE To investigate and analyze the protective effect of Huayu Jiedu Decoction(HYJD)on the inflammatory injury of cardiomyocytes induced by bacterial lipopolysaccharide(LPS),and its molecular mecha-nism of inhibitory effect on inflammatory response.METHODS H9c2 cells were cultured in vitro and divided into:the blank control group(Control group),the model control group(LPS group),the drug treatment group(HYJD group)and the combined treatment group(LPS+HYJD group).H9c2 cells were treated with different concentrations of HYJD(2.5,5,10,20 and 40 mg/ml)for 24 h,and the activity of H9c2 cells was detected by MTT assay.Additionally,H9c2 cells were treated with LPS-induced myocardial inflammatory injury cell model after 24 h of HYJD intervention at each concentration gradients to detect the cell proliferation changes,as well as to detect the levels of apoptosis of cardiomyocytes and the levels of interleukin(IL)-1β,IL-6,IL-8,IL-10 and tumor necrosis factor(TNF)-α in the culture supernatant of experimental groups,the changes in the protein ex-pression of NO production and the expression changes of iNOS and TLR4/NF-κB signaling pathway protein,and Real-time fluorescence quantitative PCR(qRT-PCR)was used to detect mRNA expression of IL-7R,P38(MAPK)and CXCR2.RESULTS Compared with the Control group,low-concentration HYJD had no significant effect on H9c2 cell viability and did not induce cytotoxic effect,and HYJD increased the survival rate of H9c2 cells in the LPS-induced myocardial inflammatory injury model,and effectively reversed the inhibitory effect of the pro-liferative activity of H9c2 cells induced by LPS.Compared with the control group,the difference in apoptosis level of H9c2 cells in the HYJD monotherapy group was not statistically significant,while the levels of inflammatory apoptosis of H9c2 cells induced by LPS was elevated(P＜0.05).Compared with the LPS group,HYJD inhibited the levels of inflammatory apoptosis in H9c2 cells induced by LPS(P＜0.05),reduced the production of pro-in-flammatory cytokines such as IL-1β,IL-6,IL-8 and TNF-α in the supernatant of the LPS-induced myocardial in-flammatory injury H9c2 cell culture,and upregulated the anti-inflammatory cytokine IL-10.Additionally,com-pared with the Control group,the LPS group showed an increased level of NO release(P＜0.05),while the difference in NO release in the low-concentration(5 mg/ml)HYJD was not statistically significant.Compared with the LPS group,the NO release levels in each HYJD intervention group showed a concentration-dependent de-crease(all P＜0.05).Furthermore,compared with the control group,whereas the expression levels of iNOS and TLR4/NF-κB signaling pathway proteins in the LPS-induced H9c2 cells were both elevated(P＜0.05).CONCLUSION HYJD exhibits protective effects against LPS-induced septic myocardial injury and can exert an in-hibitory effect on inflammatory response,and the molecular mechanisms may be related to the inhibition of the ac-tivation of the TLR4/NF-κB signaling pathway and the down-regulation of the expression of inflammatory genes,etc.,and it may have a good biological activity in the prevention and treatment of septic myocardial injury.

Objective:To evaluate the elbow joint function of patients with endemic skeletal fluorosis based on Mayo elbow joint function score and to study its application value.Methods:Based on the monitoring results of drinking water-borne endemic fluorosis in Gansu Province from 2018 to 2023, and the results of the 2019 survey on the current cases of endemic diseases, 8 typical endemic counties were selected from 48 counties with drinking water-borne endemic fluorosis. A cluster sampling method was used to investigate all adult skeletal fluorosis patients over 25 years old in the endemic fluorosis villages under the jurisdiction of the 8 counties. A total of 1 096 skeletal fluorosis patients were included as study subjects. By extracting demographic basic information, follow-up records, elbow joint X-ray films, diagnostic information, and the Mayo elbow joint function score scale of patients, a retrospective analysis was conducted on the distribution characteristics of the Mayo elbow joint function score in different genders, ages, clinical grades, disease duration, and Kellgren-Lawrence (K-L) grading of osteoarthritis.Results:(1) The distribution of total scores on the scale: 638 cases (58.21%) were rated as "poor" (< 60 points), 345 cases (31.48%) were rated as "fair" (60 - 74 points), 96 cases (8.76%) were rated as "good" (75 - 89 points), and 17 cases (1.55%) were rated as "excellent" (≥90 points). There were statistically significant differences in the total score grading of the scale among patients with different genders, ages, clinical grades, disease duration, and K-L grading of osteoarthritis (χ 2 = 44.90, 147.20, 264.41, 211.96, 262.85, P < 0.001). (2) The scores of each dimension of the scale: the joint stability dimension scores were compared among different ages, clinical grades, disease duration, and K-L grading of osteoarthritis, and the differences were statistically significant ( P < 0.05). The scores of joint pain, range of motion, and daily living ability dimensions were compared among different genders, ages, clinical grades, disease duration, and K-L grading of osteoarthritis, and the differences were statistically significant ( P < 0.05). Conclusions:Based on the Mayo elbow joint function score scale, patients with endemic skeletal fluorosis have relatively poor elbow joint function. The scores of each dimension of the scale are influenced by factors such as genders, ages, clinical grades, disease duration, and joint degeneration. The distribution of scale scores is consistent with the clinical reality of endemic skeletal fluorosis and has certain application value in assessing disease condition.

This study aims to establish a convenient,new and visual detection method for the field diagnosis of Pasteurella multocida(Pm).With reference to the Pm kmt1 gene conserved sequence published in GenBank,PCR amplification primers were designed,the amplified kmt1 gene was cloned into pMD19-T vector,and the recombinant plasmid standard pMD19-T-kmt 1 was estab-lished and identified by PCR and sequencing.Using pMD1 9-T-kmt 1 plasmid as template and kmt1 gene as target gene,basic primers were designed and synthesized.According to the requirements of LFD,a probe(Pm-P)was designed,and the RPA-LFD method for Pm detection was established by optimizing the reaction conditions.Specificity and sensitivity tests were carried out,and 64 clini-cal samples were tested by the method.The results showed that the established Pm RPA-LFD method could be amplified at 37 ℃ for 15 min.Escherichia coli(E.coli),Salmonella enteriditis(SE),Riemerella anatipestifer(RA),Staphylococcus,goose parvovirus(GPV),duck plague virus(DPV),Muscovy duck parvovirus(MDPV)DNA was extracted as the template,and plasmid standard pMD19-T-kmt 1 was used as the positive control.All the positive controls were negative,indicating that the method had good specificity.The plasmid standard pMD1 9-T-kmt 1 was diluted with a 10-fold ratio,and the plasmid standard with a concentration of 107-100 copies/μL was used as the template.The sensitivity was 1.50×101 copies/μ,,which was 100 times higher than that of PCR.A total of 64 clinical samples with suspected RA were subjected to testing using PCR,RPA and LAMP-LFD,with a 100％compliance rate for all three detection tests.The results show that the established RPA-LFD method has the characteristics of strong specificity,high sensitivity,fast speed and visualization,and can be applied to the field detection of Pm.

OBJECTIVE To investigate and analyze the protective effect of Huayu Jiedu Decoction(HYJD)on the inflammatory injury of cardiomyocytes induced by bacterial lipopolysaccharide(LPS),and its molecular mecha-nism of inhibitory effect on inflammatory response.METHODS H9c2 cells were cultured in vitro and divided into:the blank control group(Control group),the model control group(LPS group),the drug treatment group(HYJD group)and the combined treatment group(LPS+HYJD group).H9c2 cells were treated with different concentrations of HYJD(2.5,5,10,20 and 40 mg/ml)for 24 h,and the activity of H9c2 cells was detected by MTT assay.Additionally,H9c2 cells were treated with LPS-induced myocardial inflammatory injury cell model after 24 h of HYJD intervention at each concentration gradients to detect the cell proliferation changes,as well as to detect the levels of apoptosis of cardiomyocytes and the levels of interleukin(IL)-1β,IL-6,IL-8,IL-10 and tumor necrosis factor(TNF)-α in the culture supernatant of experimental groups,the changes in the protein ex-pression of NO production and the expression changes of iNOS and TLR4/NF-κB signaling pathway protein,and Real-time fluorescence quantitative PCR(qRT-PCR)was used to detect mRNA expression of IL-7R,P38(MAPK)and CXCR2.RESULTS Compared with the Control group,low-concentration HYJD had no significant effect on H9c2 cell viability and did not induce cytotoxic effect,and HYJD increased the survival rate of H9c2 cells in the LPS-induced myocardial inflammatory injury model,and effectively reversed the inhibitory effect of the pro-liferative activity of H9c2 cells induced by LPS.Compared with the control group,the difference in apoptosis level of H9c2 cells in the HYJD monotherapy group was not statistically significant,while the levels of inflammatory apoptosis of H9c2 cells induced by LPS was elevated(P＜0.05).Compared with the LPS group,HYJD inhibited the levels of inflammatory apoptosis in H9c2 cells induced by LPS(P＜0.05),reduced the production of pro-in-flammatory cytokines such as IL-1β,IL-6,IL-8 and TNF-α in the supernatant of the LPS-induced myocardial in-flammatory injury H9c2 cell culture,and upregulated the anti-inflammatory cytokine IL-10.Additionally,com-pared with the Control group,the LPS group showed an increased level of NO release(P＜0.05),while the difference in NO release in the low-concentration(5 mg/ml)HYJD was not statistically significant.Compared with the LPS group,the NO release levels in each HYJD intervention group showed a concentration-dependent de-crease(all P＜0.05).Furthermore,compared with the control group,whereas the expression levels of iNOS and TLR4/NF-κB signaling pathway proteins in the LPS-induced H9c2 cells were both elevated(P＜0.05).CONCLUSION HYJD exhibits protective effects against LPS-induced septic myocardial injury and can exert an in-hibitory effect on inflammatory response,and the molecular mechanisms may be related to the inhibition of the ac-tivation of the TLR4/NF-κB signaling pathway and the down-regulation of the expression of inflammatory genes,etc.,and it may have a good biological activity in the prevention and treatment of septic myocardial injury.

Objective:This study based on comprehensive bioinformatics technology aimed to investigate the pathogenesis of ferroptosis in IgA nephropathy(IgAN)from an immunological perspective,and to identify the key genes and functional pathways.Methods:Differentially expressed ferroptosis-related genes(DE-FRGs)were identified from GSE93798 and analyzed by Kyoto Ency-clopedia of Genes and Genomes(KEGG)and Gene Ontology(GO).STRING and Cytoscape were used to construct a protein-protein interaction(PPI)network.Degree and MCODE algorithm were run.Combining LASSO regression,SVM-RFE and PLS-DA machine learning to construct the optimal feature selection hub gene.Cibersort and ssGESA algorithms were used to assess the immune infiltra-tion,as well as to explore the relationship between hub genes and immune infiltration.Single cell RNA sequence(scRNA-seq)data was used to analyze the location of hub genes in IgAN cell populations.Gene Set Enrichment Analysis(GSEA)was performed on hub genes.The HPA database was used to obtain the position of the specific protein of the hub genes,and the DsigDB database was used to predict the target drug.The GSE37460,GSE116626 and GSE73953 were treated as validation sets and diagnostic effectiveness evalua-tion.Collecting IgAN and healthy kidney tissue for immunohistochemical testing of hub genes expression.Results:This study identi-fied 94 DE-FRGs and 3 hub genes(JJUN,EGR1 and DDR2).KEGG and GO analysis indicated that the DE-FRGs were mainly con-centrated in pathways of ferroptosis,reactive oxygen species,responsing to oxidative stress and metal particles,mitochondrion,and transcriptional regulatory complex.GESA analysis involved in amino acid and fatty acid metabolism.The analysis of immune cell infil-tration in IgAN revealed an increased ratio of naive B cells,mast cells,and CD4+T cells.Differential analysis of immune function re-vealed that mechanisms such as chemokine receptor signaling pathways,human leukocyte antigen signaling pathways,and inflamma-tion promotion were more active in IgAN.Furthermore,the correlation was observed between immune infiltration and hub genes.ScRNA-seq analysis demonstrated that hub genes were localized in monocytes,macrophages,poximal convoluted tubule cells,princi-pal cells,and smooth muscle cells.Validations set analysis suggested that JUN and SIRT1 had a diagnostic value.The HPA database analysis showed that JUN was mainly located in the nucleus,while EGR1 was located in the membrane and cytoplasm.The 2 278 po-tential drugs for the treatment of IgAN were predicted.Finally,the results of immunohistochemistry and bioinformatics analysis were consistent.Conclusion:Ferroptosis may be associated with immune cell infiltration and immune related function during the occur-rence and development of IgAN.

Objective To establish and validate the reference interval for serum anti-Müllerian hormone(AMH)in healthy women of childbearing age in Shenzhen using an indirect method based on mathematical statistics.Methods Collected the AMH data for women aged 21～50 in outpatient and physical examination populations in Shenzhen Maternity and Child Health Care Hospital from 2017 to 2023.Grouping by age range of five,first performed normality tested on each group of data,and used the interquartile range method to remove outliers for non-normal data.Then established reference intervals for AMH in different age groups through indirect method(Hoffmann method)and verified them.Compared with the reference interval of the reagent instructions and analyzed the correlation be-tween serum AMH levels and age.Results The correlation coefficient between serum AMH and age in women of childbearing age was-0.642,and the difference was statistically significant(P<0.001).The distribution of AMH levels among women in six age groups was compared,and the difference was statistically significant(H=28 392.655,P<0.05),and AMH levels showed a decreasing trend with age.The reference range of serum AMH at ages 21～25,26～30,31～35,36～40,41～45 and 46～50 yesrs were 0.92～11.30,0.68～9.43,0.38～7.51,0.12～6.93,<4.42 and<1.83ng/ml,respectively,and all reference intervals in each group had been validated.Compared to the age groups provided by the manufacturer,the age groups in this study are more refined,the reference range was narrower,and the interpretation of clinical and laboratory data was more accurate.Conclusion This study used Hoffmann's indirect method for the first time to establish a reference range for AMH in women aged 21～50 years in Shenzhen.The reference interval established is more in line with the actual situation and is a simple and reliable acquisition mode,suitable for promotion and wide application in clinical labo-ratories.

Objective To establish and validate the reference interval for serum anti-Müllerian hormone(AMH)in healthy women of childbearing age in Shenzhen using an indirect method based on mathematical statistics.Methods Collected the AMH data for women aged 21～50 in outpatient and physical examination populations in Shenzhen Maternity and Child Health Care Hospital from 2017 to 2023.Grouping by age range of five,first performed normality tested on each group of data,and used the interquartile range method to remove outliers for non-normal data.Then established reference intervals for AMH in different age groups through indirect method(Hoffmann method)and verified them.Compared with the reference interval of the reagent instructions and analyzed the correlation be-tween serum AMH levels and age.Results The correlation coefficient between serum AMH and age in women of childbearing age was-0.642,and the difference was statistically significant(P<0.001).The distribution of AMH levels among women in six age groups was compared,and the difference was statistically significant(H=28 392.655,P<0.05),and AMH levels showed a decreasing trend with age.The reference range of serum AMH at ages 21～25,26～30,31～35,36～40,41～45 and 46～50 yesrs were 0.92～11.30,0.68～9.43,0.38～7.51,0.12～6.93,<4.42 and<1.83ng/ml,respectively,and all reference intervals in each group had been validated.Compared to the age groups provided by the manufacturer,the age groups in this study are more refined,the reference range was narrower,and the interpretation of clinical and laboratory data was more accurate.Conclusion This study used Hoffmann's indirect method for the first time to establish a reference range for AMH in women aged 21～50 years in Shenzhen.The reference interval established is more in line with the actual situation and is a simple and reliable acquisition mode,suitable for promotion and wide application in clinical labo-ratories.

Background: En bloc resection is recommended for the treatment of malignant and aggressive benign spinal tumors; however, it often requires a combined anterior-posterior approach, which is usually accompanied by longer surgical duration, increased blood loss, larger trauma, and surgical complexity. The present study describes a novel rotation-reversion technique for en bloc resection of the thoracic and upper lumbar spinal tumors using a posterior-only approach and evaluate its safety and efficacy. Methods: Thirteen patients with thoracic and upper lumbar (L1-L3) spinal tumors were treated with en bloc resection using the rotation-reversion technique through a posterior-only approach at our institution between 2015 and 2023. The clinical characteristics and surgical results of the patients were reviewed and analyzed. Results: Posterior-only en bloc resection was performed successfully in all 13 patients using the rotation-reversion technique, with a median follow-up of 30.4 months (range, 6–74 months). The average maximum size of these 13 tumors was 5.7 × 5.8 × 4.8 cm.The mean operation time and blood loss were 458.5 minutes (range, 220–880 minutes) and 3,146.2 mL (range, 1,000–6,000 mL), respectively, with 4 of the 13 patients (30.8%) experiencing perioperative complications. Negative margins were achieved in all the 13 patients (100%). One patient experienced local recurrence (7.7%) and 1 patient experienced instrumentation failures. Interbody fusion was confirmed in 11 of the 13 patients (84.6%), with a median fusion time of 6.9 months. All of the 13 patients experienced varying degrees of mild postoperative neurological deficits owing to resection of the nerve roots affected by tumor invasion of the vertebrae. No vessel injury or postoperative neurological paralysis occurred, except 1 patient who had been completely paralyzed before surgery. Conclusions The rotation-reversion technique is an effective procedure for en bloc resection of selected thoracic and upper lumbar spinal tumors through the posterior-only approach.

Background: En bloc resection is recommended for the treatment of malignant and aggressive benign spinal tumors; however, it often requires a combined anterior-posterior approach, which is usually accompanied by longer surgical duration, increased blood loss, larger trauma, and surgical complexity. The present study describes a novel rotation-reversion technique for en bloc resection of the thoracic and upper lumbar spinal tumors using a posterior-only approach and evaluate its safety and efficacy. Methods: Thirteen patients with thoracic and upper lumbar (L1-L3) spinal tumors were treated with en bloc resection using the rotation-reversion technique through a posterior-only approach at our institution between 2015 and 2023. The clinical characteristics and surgical results of the patients were reviewed and analyzed. Results: Posterior-only en bloc resection was performed successfully in all 13 patients using the rotation-reversion technique, with a median follow-up of 30.4 months (range, 6–74 months). The average maximum size of these 13 tumors was 5.7 × 5.8 × 4.8 cm.The mean operation time and blood loss were 458.5 minutes (range, 220–880 minutes) and 3,146.2 mL (range, 1,000–6,000 mL), respectively, with 4 of the 13 patients (30.8%) experiencing perioperative complications. Negative margins were achieved in all the 13 patients (100%). One patient experienced local recurrence (7.7%) and 1 patient experienced instrumentation failures. Interbody fusion was confirmed in 11 of the 13 patients (84.6%), with a median fusion time of 6.9 months. All of the 13 patients experienced varying degrees of mild postoperative neurological deficits owing to resection of the nerve roots affected by tumor invasion of the vertebrae. No vessel injury or postoperative neurological paralysis occurred, except 1 patient who had been completely paralyzed before surgery. Conclusions The rotation-reversion technique is an effective procedure for en bloc resection of selected thoracic and upper lumbar spinal tumors through the posterior-only approach.

Background: En bloc resection is recommended for the treatment of malignant and aggressive benign spinal tumors; however, it often requires a combined anterior-posterior approach, which is usually accompanied by longer surgical duration, increased blood loss, larger trauma, and surgical complexity. The present study describes a novel rotation-reversion technique for en bloc resection of the thoracic and upper lumbar spinal tumors using a posterior-only approach and evaluate its safety and efficacy. Methods: Thirteen patients with thoracic and upper lumbar (L1-L3) spinal tumors were treated with en bloc resection using the rotation-reversion technique through a posterior-only approach at our institution between 2015 and 2023. The clinical characteristics and surgical results of the patients were reviewed and analyzed. Results: Posterior-only en bloc resection was performed successfully in all 13 patients using the rotation-reversion technique, with a median follow-up of 30.4 months (range, 6–74 months). The average maximum size of these 13 tumors was 5.7 × 5.8 × 4.8 cm.The mean operation time and blood loss were 458.5 minutes (range, 220–880 minutes) and 3,146.2 mL (range, 1,000–6,000 mL), respectively, with 4 of the 13 patients (30.8%) experiencing perioperative complications. Negative margins were achieved in all the 13 patients (100%). One patient experienced local recurrence (7.7%) and 1 patient experienced instrumentation failures. Interbody fusion was confirmed in 11 of the 13 patients (84.6%), with a median fusion time of 6.9 months. All of the 13 patients experienced varying degrees of mild postoperative neurological deficits owing to resection of the nerve roots affected by tumor invasion of the vertebrae. No vessel injury or postoperative neurological paralysis occurred, except 1 patient who had been completely paralyzed before surgery. Conclusions The rotation-reversion technique is an effective procedure for en bloc resection of selected thoracic and upper lumbar spinal tumors through the posterior-only approach.