Objective To study the effect of Mycobacterium tuberculosis (Mtb) Pro-Pro-Glu-59 (PPE59) protein on the biological function of Mycobacterium smegmatis (Ms) and the regulation of host cell immune response. Methods PPE59 gene fragment was obtained by PCR amplification, cloned into pALACE, constructed into recombinant pALACE-PPE59 vector, and electro-transformed into Ms. Western blot was applied to analyse PPE59 expression and subcellular localization. The survival of Ms_Vec and Ms_PPE59 under low acid (pH=3 and pH=5) conditions and active surface pressure sodium dodecyl sulfate (SDS) conditions and their intracellular survival in macrophages were analyzed. ELISA was used to detect the cytokine (IL-1β, IL-6, IL-12, TNF-α and IL-10) expression levels of Ms_Vec and Ms_PPE59 infected macrophages. Results PPE59 protein localized to the cell wall of Ms can enhance the acid-resistance and anti-SDS effect of Ms, which is conducive to the survival of Ms in macrophages. PPE59 significantly decreased the secretion levels of pro-inflammatory cytokines (IL-1β, IL-6, IL-12 and TNF-α), and promoted the secretion levels of anti-inflammatory cytokine (IL-10). Conclusion PPE59 enhances the survival ability of Ms under low acid and SDS pressure and promotes its intracellular survival by regulating the cytokine secretion levels.
Mycobacterium smegmatis/metabolism* ; Macrophages/metabolism* ; Cytokines/metabolism* ; Mycobacterium tuberculosis/metabolism* ; Bacterial Proteins/metabolism* ; Animals ; Mice ; Antigens, Bacterial/metabolism*

Varicocele (VC) is a common cause of male infertility, yet there is a lack of molecular information for VC-associated male infertility. This study investigated alterations in the seminal plasma metabolomic and lipidomic profiles of infertile male VC patients. Twenty infertile males with VC and twenty-three age-matched healthy controls (HCs) were recruited from Peking Union Medical College Hospital (Beijing, China) between October 2019 and April 2021. Untargeted metabolite and lipid profiles from seminal plasma were analyzed using mass spectrometry. Four hundred and seventy-six metabolites and seventeen lipids were significantly different in infertile male VC patients compared to HCs. The top enriched pathways among these significantly different metabolites are protein digestion and absorption, aminoacyl-transfer RNA (tRNA) biosynthesis, and biosynthesis of amino acids. Different key lipid species, including triglyceride (TG), diacylglycerol (DG), ceramides (Cer), and phosphatidylserine (PS), varied between VC and HC groups. The distinct metabolites and lipids were moderately correlated. DL-3-phenyllactic acid is a potential diagnostic biomarker for VC-related male infertility (area under the curve [AUC] = 0.893), positively correlating with sperm count, concentration, and motility. Furthermore, DL-3-phenyllactic acid is the only metabolite shared by all four comparisons (VC vs HC, VC-induced oligoasthenospermia [OAS] vs VC-induced asthenospermia [AS], OAS vs HC, and AS vs HC). DL-3-phenyllactic acid significantly decreased in OAS than AS. Metabolite-targeting gene analysis revealed carbonic anhydrase 9 (CA9) might be the strongest candidate associated with the onset and severity of VC. The seminal plasma metabolite and lipid profiles of infertile males with VC differ significantly from those of HCs. DL-3-phenyllactic acid could be a promising biomarker.
Humans ; Male ; Varicocele/complications* ; Infertility, Male/etiology* ; Semen/metabolism* ; Lipidomics ; Adult ; Metabolomics ; Case-Control Studies ; Biomarkers/metabolism*

Objective To develop a portable heat illness/stroke first-aid kit for on-site first aid of heat illness patients and verify its application effect.Methods The portable heat illness/stroke first-aid kit was composed of a main box,an adjustable telescopic rod,adjustable shoulder straps,universal rollers and a thermal insulation container.The main box made of aluminum alloy material had the inner surface lined with Oxford cloth,which was equipped with an infrared cochlear thermometer,a nebulizer,first aid medicines,heat stroke medicines,a sun umbrella,a cooling blanket,etc;the adjustable telescopic rod was made of aluminum-magnesium alloy;the adjustable shoulder straps was made of high-density nylon webbing;the universal rollers were rubberized and had wheel brakes;the thermal insulation container was located in the lower storage compartment inside the main box,which used polyurethane(PU)material for thermal insulation.The data on on-site first aid of the patients with moderate heat illness or stroke at some institution from 2019 to 2022 were analyzed retrospectively,with the patients treated with the traditional methods enrolled into a control group and the ones with the first-aid kit into an experimental group.The temperature changes at 0,10,20,30,40,50 and 60 min after the start of treatment were investigated to compare the cooling effects of the two groups.Results The heat illness/stroke first-aid kit lowered the patient temperature effectively during the on-site first aid of the patients with moderate heat illness or stroke,with higher cooling speed and effect than the traditional methods,especially at the early stage of treatment.Conclusion The heat illness/stroke first-aid kit with complete functions and easy operation decreases temperature of heat illness patients efficiently,and can be applicable to the scenarios such as field training and outdoors high-temperature operation.[Chinese Medical Equipment Journal,2025,46(9):108-113]

Aim To evaluate the role of the glucose transporter protein 1(GLUT1)-dependent glycolytic in the attenuation of oxygen-glucose deprivation-reoxygen-ation(OGD/R)injury in HK-2 cells by dexmedetomi-dine(Dex).Methods C57/BL6 mice were random-ly divided into three groups(n=6),namely,sham operation group(Sham group),renal ischemia reper-fusion group(I/R group)and Dex group(I/R+Dex group).Serum creatinine(Cr)and urea nitrogen(BUN)were measured,while the levels of key glyco-lytic enzymes HK2,PFKFB3 and GLUT1 were meas-ured.HK-2 cells were cultured and randomised into seven groups(n=6),which was treated with OGD/R,overexpression or interference with GLUT1,Dex and glycolysis inhibitor 2-DG.CCK-8 and LDH activi-ty were used to detect cellular damage.Glycolysis lev-els were detected by lactate and ECAR.The inflamma-tory level was reflected by qRT-PCR for IL-6 and TNF-α.qRT-PCR and Western blot were performed to de-tect the levels of GLUT1,HK2,and PFKFB3.Results Dex significantly ameliorated kidney injury and HK-2 cell injury(P＜0.05).Dex inhibited the OGD/R-induced rise in lactate and extracellular acidification rate(ECAR),as evidenced by suppression of the ex-pression of GLUT1,HK2 and PFKFB3(P＜0.05).In vitro experiments showed that GLUT1 knockdown sig-nificantly improved OGD/R-induced cellular damage.Lactate,ECAR,glycolysis-related mRNAs and pro-teins were inhibited by GLUT1 knockdown(P＜0.05).Significantly,there were no significant differ-ences in above indexes after Dex treatment based on GLUT1 knockdown.Overexpression of GLUT1 abroga-ted the protective effects of Dex,while reversing the inhibitory effects of Dex on the expression of GLUT1,HK2,and PFKFB3(P＜0.05).Conclusions Dexmedetomidine attenuates OGD/R induced injury in HK-2 cells by inhibiting GLUT1-dependent glycolysis.

Objective To evaluate the effects of fasting and high-fat diet on the pharmacokinetics of rabeprazole sodium enteric-coated tablets in healthy Chinese subjects.Methods A single-center,randomized,open,two-agent,two-sequence,four-cycle,fully repeated crossover,single-dose trial design was used in this study,healthy subjects were assigned to receive single dose of rabeprazole sodium enteric-coated tablets 0.1 g in either fasting or high-fat diet state,and blood samples were taken at different time points,respectively.The concentrations of rabeprazole sodium enteric-coated in plasma were determined by liquid chromatography-tandem mass spectrometry(LC-MS/MS),the model method of the non-compartmental was used to calculate the pharmacokinetic parameters by Phoenix WinNonlin 8.2.Results The main pharmacokinetic parameters of rabeprazole sodium enteric-coated tablets in fasting state and high-fat diet state were as follows:Cmax were(339.63±156.47)and(318.86±132.13)ng·mL-1;t1/2 were(2.34±0.68)and(3.60±2.40)h;AUC0_t were(556.62±251.65)and(528.50±201.78)ng·mL-1·h;AUC0-∞ were(563.39±255.69)and(535.15±203.24)ng·mL-1·h;tmax were 3.65 and 6.99 h.After high-fat diet,the Cmax and AUC of rapeprazole sodium after high-fat and high-calorie diet decreased,Cmax decreased by 6.12％,AUC0-t decreased by 5.05％,AUC0-∞ decreased by 5.01％,andtmaxwas delayed by about 3.34 h.Cmax,AUC0-t and AUC0-∞ 90％confidence interval were 73.13％-115.10％,83.22％-112.28％and 83.40％-112.13％,respectively.Neither was between 85.00％-125.00％.Conclusion High-fat diet affects the absorption rate and degree of rabeprazole sodium enteric-coated,so it is suitable to be administered on an empty stomach.

Objective To study the plasma metabolomics of mice poisoned by different dosage of the combination of diazepam and ethanol,and to reveal the toxicological mechanisms of combined poisoning of diazepam and ethanol.Methods Female Kunming mice were randomly divided into blank group,single and combined poisoning group(n=6),Based on the LD50 of diazepam co-administered with graded ethanol doses,mice in the single-drug and combined groups received oral gavage at 1/2,1,and 2 × LD50.Retro-orbital blood samples(～500 μL)were collected within 24 hours post-administration and analyzed by UPLC-QE-MS technology.Principal component analysis and orthogonal partial least squares discriminant analysis were used to identify differential metabolites and associated metabolic pathways.Results A total of 387 differential metabolites were identified in the combined poisoning group of diazepam and ethanol implicating the key pathways including tryptophan metabolism,phenylalanine metabolism,arginine and proline metabolism,Glycerophospholipid metabolism,phenylalanine,tyrosine and tryptophan biosynthesis.Conclusion Combined diazepam and ethanol poisoning exerts significant systemic effects by disrupting neurotransmitters conduction,exacerbating oxidative stress response and dysregulating energy metabolism.

Objective:To investigate the effects of prophylactic analgesia using erector spinae plane block (ESPB) combined with ketorolac tromethamine on postoperative analgesia and early recovery of patients undergoing laparoscopic radical gastrectomy.Methods:A prospective randomized controlled study was conducted. A total of 105 gastric cancer patients who underwent laparoscopic radical gastrectomy in Heji Hospital Affiliated to Changzhi Medical College from June 2022 to October 2023 were selected, and the patients were divided into ESPB+ketorolac tromethamine group (T group), ESPB group (C1 group), and ketorolac tromethamine group (C2 group) by using the random number table method, with 35 cases in each group. The prophylactic analgesia method was preoperative ESPB + ketorolac tromethamine in T group, preoperative ESPB in C1 group and preoperative ketorolac tromethamine in C2 group. The intraoperative sufentanil and remifentanil dosage and the postoperative resting pain visual analogue scale (VAS) score, remedial analgesia, recovery status and complication occurrence were compared among the 3 groups.Results:The baseline characteristics and intraoperative conditions of the patients in the 3 groups were compared, and the differences were not statistically significant (all P > 0.05). The intraoperative sufentanil dosage in T group was less than that in C1 and C2 groups [30 (25, 30) μg vs. 35 (30, 35) μg vs. 40 (35, 45) μg], and the difference was statistically significant ( Z = 33.03, P < 0.001); the intraoperative remifentanil dosage in T group was less than that in C1 and C2 groups [0.34 (0.30, 0.40) mg vs. 0.40 (0.30, 0.50) mg vs. 0.70 (0.60, 0.85) mg], and the difference was statistically significant ( Z = 53.84, P < 0.001). The VAS scores at 1, 6, 12, 24, 48 h after surgery in T group were lower than those in C1 and C2 groups, and the differences were statistically significant (all P < 0.001). The press number of analgesic pump in T group was less than that in C1 and C2 groups [1 (0, 2) times vs. 2 (1, 2) times vs. 4 (3, 5) times], and the difference was statistically significant ( Z = 48.10, P < 0.001). The postoperative first time of exhaust and get out of bed and days of hospital stay in T group were shorter than those in C1 and C2 groups, and the differences were statistically significant (all P < 0.001). The differences in the incidence of postoperative nausea and vomiting, dizziness, pruritus, pneumonia, and anastomotic fistula among the 3 groups were not statistically significant (all P > 0.05). None of the 3 groups experienced postoperative drowsiness, hypotension or respiratory depression. Conclusions:Prophylactic analgesia using ESPB combined with ketorolac tromethamine can effectively reduce the postoperative pain of patients undergoing laparoscopic radical gastrectomy, reduce the use of analgesic drugs, and promote the early recovery of the patients in the postoperative period without increasing the postoperative complications.

Objective To analyze the imaging features of cerebral small vessel disease in patients with type 2 diabetes mellitus by multimodal MRI.Methods The clinical data of 160 patients with cerebral small vessel disease admitted to our hospital from January to December 2020 were retrospectively analyzed.According to whether they were complicated with type 2 diabetes mellitus,they were divided into the diabetic group and the non-diabetic group,with 80 cases in each group.Both groups underwent multimodal MRI scans.And the severity of lacunar infarction,the severity of subcortical and periventricular white matter lesions,white matter integral and cerebral microbleeds of patients in the two groups were compared.Results The severity of lacunar infarction(χ2=34.076,P=0.001),subcortical white matter lesions(χ2=25.000,P=0.001),periventricular white matter lesions(χ2=22.895,P=0.001)and white matter integral(t=12.370,P=0.001)of patients in the diabetic group were significantly higher than those in the non-diabetic group.No cerebral microbleeds were detected in either group of patients.Conclusion Patients with cerebral small vessel disease and type 2 diabetes mellitus show characteristic multimodal MRI changes.The increase in the number of lacunar infarction lesions and the aggravation of white matter lesions can be used as the characteristic imaging basis for the diagnosis of type 2 diabetes mellitus related cerebral small vessel disease.

Background/Aims: Metabolic dysfunction-associated steatohepatitis (MASH) is a significant risk factor for gallstone formation, but mechanisms underlying MASH-related gallstone formation remain unclear. Golgi membrane protein 1 (GOLM1) participates in hepatic cholesterol metabolism and is upregulated in MASH. Here, we aimed to explore the role of GOLM1 in MASH-related gallstone formation. Methods: The UK Biobank cohort was used for etiological analysis. GOLM1 knockout (GOLM1-/-) and wild-type (WT) mice were fed with a high-fat diet (HFD). Livers were excised for histology and immunohistochemistry analysis. Gallbladders were collected to calculate incidence of cholesterol gallstones (CGSs). Biles were collected for biliary lipid analysis. HepG2 cells were used to explore underlying mechanisms. Human liver samples were used for clinical validation. Results: MASH patients had a greater risk of cholelithiasis. All HFD-fed mice developed MASH, and the incidence of gallstones was 16.7% and 75.0% in GOLM1-/- and WT mice, respectively. GOLM1-/- decreased biliary cholesterol concentration and output. In vivo and in vitro assays confirmed that GOLM1 facilitated cholesterol efflux through upregulating ATP binding cassette transporter subfamily G member 5 (ABCG5). Mechanistically, GOLM1 translocated into nucleus to promote osteopontin (OPN) transcription, thus stimulating ABCG5-mediated cholesterol efflux. Moreover, GOLM1 was upregulated by interleukin-1β (IL-1β) in a dose-dependent manner. Finally, we confirmed that IL-1β, GOLM1, OPN, and ABCG5 were enhanced in livers of MASH patients with CGSs. Conclusions In MASH livers, upregulation of GOLM1 by IL-1β increases ABCG5-mediated cholesterol efflux in an OPN-dependent manner, promoting CGS formation. GOLM1 has the potential to be a molecular hub interconnecting MASH and CGSs.

Objective To explore the effects of Emotional Freedom Techniques combined with PERMA model intervention on fear of disease progression and quality of life in postoperative liver cancer patients.Methods A total of 58 liver cancer patients admitted from September 2023 to February 2024 were randomly divided into the experimental group(n=29)and the control group(n=29)using a random number table method.The experimental group received Emotional Freedom Techniques combined with PERMA-based positive psychological intervention,while the control group received the holistic nursing and routine psychological guidance.Both groups underwent the continuous intervention for 8 weeks.Outcomes were assessed using the Fear of Progression Questionnaire-Short Form(FoP-Q-SF),Distress Thermometer(DT),Self-Rating Anxiety Scale(SAS),Self-Rating Depression Scale(SDS),Cancer Fatigue Scale(CFS),Athens Insomnia Scale(AIS)and Index of Well-Being(IWB).Results After the intervention,the experimental group showed the significantly lower scores than those in the control group in FoP-Q-SF(24.97±2.34/34.10±8.46),DT(2.24±0.95/2.93±1.33),SAS(49.55±1.35/58.55±4.60),SDS(51.86±1.58/61.79±4.00),CFS(4.03±0.94/5.10±1.18),and AIS(2.45±1.12/4.07±0.96)(all P<0.05),while the IWB score(10.56±1.74/7.39±2.05)was significantly higher than that in the control group(P<0.05).Conclusion The combination of Emotional Freedom Techniques and PERMA model intervention can effectively alleviate the disease-related fears,improve the negative emotions,reduce the physical symptoms and enhance the subjective well-being in postoperative liver cancer patients,and it has a significant clinical application value.