Neuropathic pain, a debilitating condition caused by dysfunction of the somatosensory nervous system, remains difficult to treat due to limited understanding of its molecular mechanisms. Bioinformatics analysis identified cerebellin 2 (CBLN2) as highly enriched in human and murine proprioceptive and nociceptive neurons. We found that CBLN2 expression is persistently upregulated in dorsal root ganglia (DRG) following spinal nerve ligation (SNL) in mice. In addition, transcription factor SOX11 binds to 12 cis-regulatory elements within the Cbln2 promoter to enhance its transcription. SNL also induced SOX11 upregulation, with SOX11 and CBLN2 co-localized in nociceptive neurons. The siRNA-mediated knockdown of Sox11 or Cbln2 attenuated SNL-induced mechanical allodynia and thermal hyperalgesia. High-throughput sequencing of DRG following intrathecal injection of CBLN2 revealed widespread gene expression changes, including upregulation of numerous NF-κB downstream targets. Consistently, CBLN2 activated NF-κB signaling, and inhibition with pyrrolidine dithiocarbamate reduced CBLN2-induced pain hypersensitivity, proinflammatory cytokines and chemokines production, and neuronal hyperexcitability. Together, these findings identified the SOX11/CBLN2/NF-κB axis as a critical mediator of neuropathic pain and a promising target for therapeutic intervention.
Animals ; Neuralgia/metabolism* ; Ganglia, Spinal/metabolism* ; Up-Regulation ; Mice ; NF-kappa B/metabolism* ; SOXC Transcription Factors/genetics* ; Male ; Neuroinflammatory Diseases/metabolism* ; Mice, Inbred C57BL ; Nerve Tissue Proteins/genetics* ; Hyperalgesia/metabolism* ; Signal Transduction ; Spinal Nerves

OBJECTIVE: CRISPR-Cas protects bacteria from exogenous DNA invasion and is associated with bacterial biofilm formation and pathogenicity. METHODS: We analyzed the type I-F CRISPR-Cas system of Legionella pneumophila WX48, including Cas1, Cas2-Cas3, Csy1, Csy2, Csy3, and Cas6f, along with downstream CRISPR arrays. We explored the effects of the CRISPR-Cas system on the in vitro growth, biofilm-forming ability, and pathogenicity of L. pneumophila through constructing gene deletion mutants. RESULTS: The type I-F CRISPR-Cas system did not affect the in vitro growth of wild-type or mutant strains. The biofilm formation and intracellular proliferation of the mutant strains were weaker than those of the wild type owing to the regulation of type IV pili and Dot/Icm type IV secretion systems. In particular, Cas6f deletion strongly inhibited these processes. CONCLUSION The type I-F CRISPR-Cas system may reduce biofilm formation and intracellular proliferation in L. pneumophila.
Legionella pneumophila/pathogenicity* ; CRISPR-Cas Systems ; Biofilms/growth & development* ; Phenotype ; Bacterial Proteins/metabolism* ; Gene Deletion

This study explores whether the current external quality assessment (EQA) level and acceptable bias for basic semen analysis in China are clinically useful. We collected data of semen EQA from Andrology laboratories in the Hunan Province (China) in 2022 and searched for data in the published literature from January 2000 to December 2023 in China. On the basis of these data, we analyzed the coefficients of variation and acceptable biases of different quality control materials for basic semen analysis through robust statistics. We compared these findings with quality specifications based on biological variation from optimal, desirable, and minimum levels of bias to seek a unified and more suitable semen EQA bias evaluation standard for China's national conditions. Different sources of semen quality control material exhibited considerable variation in acceptable biases among laboratories, ranging from 8.2% to 56.9%. A total of 50.0% of the laboratories met the minimum quality specifications for progressive motility (PR), whereas 100.0% and 75.0% of laboratories met only the minimum quality specifications for sperm concentration and total motility (nonprogressive [NP] + PR), respectively. The Z value for sperm concentration and PR+NP was equivalent to the desirable performance specification, whereas the Z value for PR was equivalent only to the minimum performance specification. This study highlights the feasibility of operating external quality assessment schemes for basic semen analysis using quality specifications based on biological variation. These specifications should be unified among external quality control (EQC) centers based on biological variation.
Semen Analysis/standards* ; Humans ; China ; Male ; Quality Control ; Sperm Motility ; Sperm Count/standards*

Objective To analyze the detection rate,distribution characteristics,resistance mechanism,and mo-lecular typing of carbapenem-resistant Citrobacter freundii(CRCF),providing theoretical basis for clinical treat-ment and prevention.Methods Antimicrobial susceptibility testing results of CRCF isolated from Quanzhou First Hospital Affiliated to Fujian Medical University from 2020 to 2023 were analyzed.Resistance phenotypes of the strains were verified;resistance genes were amplified;strain homology was analyzed by multi-locus sequencing ty-ping(MLST).Whole genome sequencing analysis was performed on CF17 strains carrying both blaVIM-1,and blaNDM-1.Results A total of 119 strains of non-repetitive Citrobacter freundii were detected,27 strains were CRCF,accounting for 22.7％.CRCF was mainly isolated from departments of neurosurgery and urology,and the specimen type was mainly midstream urine.Strains were resistant to both carbapenems and cephalosporins,less resistant to amikacin,susceptible to both tigecycline and polymyxin,and with a resistance rate of＞50％to other antimicrobial agents.22 CRCF strains were successfully recovered and all were positive for antimicrobial resistance phenotype and genes.17 strains(77.3％)carried single resistance genes,including 14 strains carrying blaNDM-1,2 strains carrying blaKPC-2,and 1 strain carrying blaNDM-5.5 strains(22.7％)carried dual antimicrobial resistance genes,including strains carrying both blaKPC-2 and blaNDM-1(n=3),strain carrying both blaNDM-1 and blaIMP(n=1),and strain carrying both blaVIM-1 and blaNDM-1(n=1).A total of 8 ST types were detected,with 8 and 6 strains being ST1 16 and ST532,respectively.CF17 was ST1 16,carrying blaVIM-1 and blaNDM-1 on different plasmids,namely IncA and IncX3,respectively.Conclusion CRCF strains are mostly multidrug-resistant,with the presence of blaNDM-1 be-ing the major resistance mechanism.Some strains also carry dual resistance genes,mainly ST116 and ST532.CF17 is currently the first isolated and reported Citrobacter freundii that carries both blaVIM-1 and blaNDM-1 resistance genes in China.

Objective To analyze the detection rate,distribution characteristics,resistance mechanism,and mo-lecular typing of carbapenem-resistant Citrobacter freundii(CRCF),providing theoretical basis for clinical treat-ment and prevention.Methods Antimicrobial susceptibility testing results of CRCF isolated from Quanzhou First Hospital Affiliated to Fujian Medical University from 2020 to 2023 were analyzed.Resistance phenotypes of the strains were verified;resistance genes were amplified;strain homology was analyzed by multi-locus sequencing ty-ping(MLST).Whole genome sequencing analysis was performed on CF17 strains carrying both blaVIM-1,and blaNDM-1.Results A total of 119 strains of non-repetitive Citrobacter freundii were detected,27 strains were CRCF,accounting for 22.7％.CRCF was mainly isolated from departments of neurosurgery and urology,and the specimen type was mainly midstream urine.Strains were resistant to both carbapenems and cephalosporins,less resistant to amikacin,susceptible to both tigecycline and polymyxin,and with a resistance rate of＞50％to other antimicrobial agents.22 CRCF strains were successfully recovered and all were positive for antimicrobial resistance phenotype and genes.17 strains(77.3％)carried single resistance genes,including 14 strains carrying blaNDM-1,2 strains carrying blaKPC-2,and 1 strain carrying blaNDM-5.5 strains(22.7％)carried dual antimicrobial resistance genes,including strains carrying both blaKPC-2 and blaNDM-1(n=3),strain carrying both blaNDM-1 and blaIMP(n=1),and strain carrying both blaVIM-1 and blaNDM-1(n=1).A total of 8 ST types were detected,with 8 and 6 strains being ST1 16 and ST532,respectively.CF17 was ST1 16,carrying blaVIM-1 and blaNDM-1 on different plasmids,namely IncA and IncX3,respectively.Conclusion CRCF strains are mostly multidrug-resistant,with the presence of blaNDM-1 be-ing the major resistance mechanism.Some strains also carry dual resistance genes,mainly ST116 and ST532.CF17 is currently the first isolated and reported Citrobacter freundii that carries both blaVIM-1 and blaNDM-1 resistance genes in China.

Objective To study the anti-reflux effect of conical gastric stump embedding in radical resection of esophageal cancer.Methods A total of 60 patients who planned to undergo radical resection of esophageal cancer in our hospital from June 2020 to June 2022 were selected as the study objects,and the patients were divided into the observation group and the control group by random number table method,with 30 cases in each group.Patients in both groups underwent laparoscopic radical resection of esophageal cancer and esophagogastric end-to-side mechanical anastomosis.The observation group adopted the conical gastric stump embedding technique after esophagogastric end-to-side mechanical anastomosis.The perioperative related indexes,postoperative complications and gastroesophageal reflux of patients in the two groups were compared.The postoperative anti-reflux effect was evaluated by reflux disease questionnaire(RDQ)score and 24-hour intraesophageal pH monitoring.Results The operation time and digestive tract reconstruction time of patients in the observation group were longer than those in the control group(P<0.05),while there was no statistically significant difference in the amount of intraoperative bleeding,the number of lymph node dissection,the first exhaust time,or the postoperative hospital stay of patients between the two groups(P>0.05).There was no statistically significant difference in the overall incidence of postoperative complications between the two groups(P>0.05).The severity of postoperative gastroesophageal reflux of patients in the observation group was lighter than that in the control group,and the difference was statistically significant(P<0.05).The RDQ score,24-hour reflux frequency,>5 minutes reflux frequency,pH<4 time,and longest reflux time of patients in the observation group was significantly lower/less/shorter than those in the control group(P<0.05).Conclusion The conical gastric stump embedding technique is safe and feasible in the radical resection of esophageal cancer.Although the operation time and digestive tract reconstruction time are slightly prolonged,it does not increase the perioperative risks,which can significantly reduce the occurrence and severity of postoperative gastroesophageal reflux of patients,and achieve a good anti-reflux effect.

Rapid turnover of the intestinal epithelium is a critical strategy to balance the uptake of nutrients and defend against environmental insults, whereas inappropriate death promotes the spread of inflammation. PPARα is highly expressed in the small intestine and regulates the absorption of dietary lipids. However, as a key mediator of inflammation, the impact of intestinal PPARα signaling on cell death pathways is unknown. Here, we show that Pparα deficiency of intestinal epithelium up-regulates necroptosis signals, disrupts the gut vascular barrier, and promotes LPS translocation into the liver. Intestinal Pparα deficiency drives age-related hepatic steatosis and aggravates hepatic fibrosis induced by a high-fat plus high-sucrose diet (HFHS). PPARα levels correlate with TRIM38 and MLKL in the human ileum. Inhibition of PPARα up-regulates necroptosis signals in the intestinal organoids triggered by TNF-α and LPS stimuli via TRIM38/TRIF and CREB3L3/MLKL pathways. Butyric acid ameliorates hepatic steatosis induced by intestinal Pparα deficiency through the inhibition of necroptosis. Our data suggest that intestinal PPARα is essential for the maintenance of microenvironmental homeostasis and the spread of inflammation via the gut-liver axis.

OBJECTIVES: To study the clinical and genetic features of Joubert syndrome (JS) in children. METHODS: A retrospective analysis was performed on the clinical data, genetic data, and follow-up data of 20 children who were diagnosed with JS in the Department of Children's Rehabilitation, the Third Affiliated Hospital of Zhengzhou University, from January 2017 to July 2022. RESULTS: Among the 20 children with JS, there were 11 boys and 9 girls. The common clinical manifestations were developmental delay (20 children, 100%), abnormal eye movement (19 children, 95%), and hypotonia (16 children, 80%), followed by abnormal respiratory rhythm in 5 children (25%) and unusual facies (including prominent forehead, low-set ears, and triangular mouth) in 3 children (15%), and no limb deformity was observed. All 20 children (100%) had the typical "molar tooth sign" and "midline cleft syndrome" on head images, and 6 children (30%) had abnormal eye examination results. Genetic testing was performed on 7 children and revealed 6 pathogenic genes, i.e., the CPLANE1, RPGRIP1L, MKS1, CC2D2A, CEP120, and AHI1 genes. CONCLUSIONS For children with developmental delay, especially those with abnormal eye movement and hypotonia, it is recommended to perform a head imaging examination to determine the presence or absence of "molar tooth sign" and "midline cleft syndrome", so as to screen for JS to avoid missed diagnosis and misdiagnosis. There are many pathogenic genes for JS, and whole-exome sequencing can assist in the diagnosis of JS.
Male ; Female ; Humans ; Child ; Cerebellum ; Abnormalities, Multiple/genetics* ; Kidney Diseases, Cystic/genetics* ; Eye Abnormalities/genetics* ; Retina ; Retrospective Studies ; Muscle Hypotonia/genetics*

Objective To establish the GC characteristic choromatogram of Huanglian Shangqing tablets(pills and granules),and to investigate the quality of the medicinal materials(Weeping Forsythia,Mentha Hyplocalyx and Herba Schizonepetae)containing volatile oil in Huanglian Shangqing tablets(pills and granules).Methods Agilient HP-5 capillary column(30 m×0.32 mm,0.25 μm)was used with temperature programming,the split ratio was 5∶1,the split flow was 5 mL·min-1,the injection volume was 1 μL,and the inlet temperature was 250℃.A hydrogen flame ionization detector was used,with the detector temperature of 260℃,and the column flow was 1 mL·min-1.The quality of samples from different manufacturers was evaluated by using the GC characteristic choromatogram of volatile oil and the identification of characteristic peaks,combined with similarity analysis and principal component analysis.Results The characteristic chromatograms of 192 batches of samples from 56 manufacturers were established.The peaks of Weeping Forsythia,Mentha Hyplocalyx and Herba Schizonepetae were identified as α-pinene and β-pinene,menthol and menthone,and pulegone respectively.The GC choromatogram of volatile oil components in Huanglian Shangqing tablets(pills)from different manufacturers were quite different,and some peaks of indicator components in the products from a few manufacturers have missing.Conclusion The established GC characteristic chromatograms can reflect the product quality status and can be used for the quality evaluation and product quality control of Huanglian Shangqing tablets(pills and capsules).Huanglian Shangqing tablets(pills and capsules)from some manufacturers are insufficient in the active ingredients of volatile oil,therefore,the product quality needs to be improved.

Idiopathic pulmonary fibrosis(IPF)is a refractory disease,which seriously endangers the health of patients and increases the mortality of patients.Its pathogenesis is still unclear,but it is mainly driven by alveolar epithelial cell injury.Epithelial-mesenchymal transition is an important part of the disease and is considered to be a potential therapeutic target.A series of in vitro and in vivo experiments have confirmed the role in IPF.This article reviews the interaction mechanisms of IPF-related cells and fibrosis factors in IPF,in order to provide a theoretical basis for further exploration of the occurrence and development mechanism of IPF and future clinical prevention and treatment.