ObjectiveTo analyze the phylogroups of Klebsiella pneumoniae isolates from laboratory rats and mice housed at the Laboratory Animal Center Fudan University, optimize the identification method for Klebsiella pneumoniae, and perform virulence genes detection and antimicrobial susceptibility testing, thereby providing a practical basis for microbial detection strategies and prevention and control measures in laboratory animal facilities. MethodsBetween September 2023 and September 2025, ileocecal contents were collected from laboratory rats and mice purchased from laboratory animal suppliers, from which 11 strains of Klebsiella pneumoniae were isolated and cryopreserved. These strains were tested for β-lactamase genes using multiplex PCR for phylogrouping. Furthermore, representative strains from different phylogroups were selected for biochemical tests, mass spectrometry analysis, 16S rRNA gene and tyrB gene sequencing, as well as virulence genes detection and antimicrobial susceptibility testing. ResultsMultiplex PCR detection of β-lactamase genes revealed that among 11 strains, 4 were identified as Klebsiella pneumoniae (Kp1), 4 as Klebsiella quasipneumoniae (Kp2), and 3 as Klebsiella variicola (Kp3). Kp1-Kp3 could not be differentiated using biochemical tests, mass spectrometry analysis, or 16S rRNA gene sequencing, whereas multiplex PCR of β-lactamase genes and tyrB gene sequencing enabled phylogrouping of all three. There were differences in the detection rates of capsular genes (capsule associated gene G and magnesium transport system ATP-binding protein A) and siderophore genes (Klebsiella pneumoniae iron uptake system genes) among Kp1-Kp3; susceptibility to different antibiotics also varied considerably. ConclusionKlebsiella pneumoniae phylogroups isolated from commercially produced laboratory rats and mice are similar to those in human clinical samples and likewise include multiple phylogroups of the species complex. In this study, the identification methods of Kp1-Kp3 were optimized,furthermore selected virulence genes and antimicrobial resistance in Kp1-Kp3 were preliminarily investigated, which could provide precise reference for the treatment of infections caused by different phylogroups occurring in laboratory animal facilities.

Objective:To analyze the registration status of clinical trials of TCM treatment for functional dyspepsia (FD); To provide references for relevant clinical trial registration and implementation.Methods:The clinical registration trials of TCM for the treatment of FD were retrieved from ChiCTR and clinicaltrials.gov until January 1st, 2025. Excel 2019 was used to extract and analyze study characteristics, including basic information (registration time, country/region, institution, funding source), study features and quality (study type, trial design, phase, number of centers, sample size, interventions, outcome measures, methodological quality, reporting quality), as well as recruitment status and ethical review.Results:A total of 89 registered studies were included. Registration began in 2008 with 3 studies, peaking in 2021 with 14 studies. The studies involved 6 countries, with domestic trials covering 18 provincial-level regions in China and 52 clinical trial institutions. Primary funding sources were national funding and pharmaceutical company support. Most studies were interventional with randomized parallel-controlled designs, commonly using simple randomization. Post-marketing drug trials were the most frequent phase, and single-center trials predominated, involving a total sample size of 19 776 cases. Common interventions included acupuncture/massage and Chinese patent medicines. Frequently used outcome measures were effectiveness rate, symptom evaluation, and quality of life assessment, though issues existed with non-standardized reporting and lack of TCM-specific indicators. Methodological quality needed further improvement.Conclusions:Although the number of registered clinical trials of TCM treatment for FD has increased, the overall quantity remains insufficient. Researchers should emphasize optimization of registration details and study protocols to further enhance registration quality.

Objective:To investigate the efficacy and safety of hydropuncture combined with concentrated growth factor (CGF) injection in improving facial aging.Methods:In this randomized controlled trial, 90 female patients with facial aging were prospectively enrolled at Plastic Surgery Hospital of Zhengzhou First People′s Hospital from January to August 2023, aged (34.0±1.5) years. Participants were randomly divided into three groups using a random number table ( n=30): hydropuncture group, CGF group, and combined hydropuncture-CGF group. All patients received monthly treatments for three sessions. Efficacy was evaluated one month after the third treatment using the VISIA skin analysis system to score pore size, skin texture, red areas, and brown spots. The clinical outcomes of facial rejuvenation and patient satisfaction were compared among the groups. Adverse reactions, including redness, pain, and allergies, were recorded. Results:Compared with the baseline data, all groups showed significant improvement in all parameters post-treatment (all P<0.01). Pretreatment scores for pores, texture, red areas, and brown spots exhibited no significant differences among groups (all P>0.05). After treatment, the combined group demonstrated the highest improvement, with significantly better scores for all parameters compared to the hydropuncture and CGF groups (all P<0.01). Patient satisfaction rates were 56.7% (17/30) in the hydropuncture group, 60.0% (18/30) in the CGF group, and 70.0% (21/30) in the combined group, with statistically significant differences ( P<0.001). No severe adverse reactions, such as infection, nodules, or hyperplasia, were observed in any group. Conclusion:Hydropuncture combined with CGF injection effectively improves facial aging with a high safety profile and no severe adverse reactions.

OBJECTIVE: To observe the clinical effect of acupuncture-moxibustion therapy of Daoqi Tongluo (conducting qi and unblocking collateral) on chronic primary tinnitus. METHODS: A total of 32 patients with chronic primary tinnitus were included and treated with the acupuncture-moxibustion therapy of Daoqi Tongluo. This regimen was composed of abdominal acupuncture, body acupuncture, warm needling and posterior-auricular local flashing cupping, Zhongwan (CV12), Guanyuan (CV6) and Yindu (KI9), Tinggong (SI19), Cong'er point, Waiguan (TE5) of the affected side, etc. are selected. The treatment was given once every two days, 3 treatments a week; and one course of intervention was required, with 10 treatments included. Before and after treatment, the scores of tinnitus handicap inventory (THI), tinnitus evaluation questionnaire (TEQ), self-rating scale of sleep (SRSS), self-rating anxiety scale (SAS), and self-rating depression scale (SDS) were observed, and the clinical effect was evaluated. RESULTS: After interventions, the scores of THI, TEQ, SRSS, SAS and SDS were reduced in comparison with those before interventions in the patients (P<0.001, P<0.01, P<0.05), and the total effective rate was 71.9% (23/32). CONCLUSION Acupuncture-moxibustion therapy of Daoqi Tongluo is effective on chronic primary tinnitus and this therapy can alleviate tinnitus degree, improve sleep quality and attenuate the anxious and depressive emotion of the patients.
Adult ; Aged ; Female ; Humans ; Male ; Middle Aged ; Young Adult ; Acupuncture Points ; Acupuncture Therapy ; Chronic Disease/therapy* ; Moxibustion ; Tinnitus/psychology* ; Treatment Outcome

Objective:To investigate the efficacy and safety of hydropuncture combined with concentrated growth factor (CGF) injection in improving facial aging.Methods:In this randomized controlled trial, 90 female patients with facial aging were prospectively enrolled at Plastic Surgery Hospital of Zhengzhou First People′s Hospital from January to August 2023, aged (34.0±1.5) years. Participants were randomly divided into three groups using a random number table ( n=30): hydropuncture group, CGF group, and combined hydropuncture-CGF group. All patients received monthly treatments for three sessions. Efficacy was evaluated one month after the third treatment using the VISIA skin analysis system to score pore size, skin texture, red areas, and brown spots. The clinical outcomes of facial rejuvenation and patient satisfaction were compared among the groups. Adverse reactions, including redness, pain, and allergies, were recorded. Results:Compared with the baseline data, all groups showed significant improvement in all parameters post-treatment (all P<0.01). Pretreatment scores for pores, texture, red areas, and brown spots exhibited no significant differences among groups (all P>0.05). After treatment, the combined group demonstrated the highest improvement, with significantly better scores for all parameters compared to the hydropuncture and CGF groups (all P<0.01). Patient satisfaction rates were 56.7% (17/30) in the hydropuncture group, 60.0% (18/30) in the CGF group, and 70.0% (21/30) in the combined group, with statistically significant differences ( P<0.001). No severe adverse reactions, such as infection, nodules, or hyperplasia, were observed in any group. Conclusion:Hydropuncture combined with CGF injection effectively improves facial aging with a high safety profile and no severe adverse reactions.

Objective:To explore the changes in serum advanced receptor for glycation end-products (sRAGE) and neuregulin-1 (NRG-1) in patients with different conditions of H-type hypertension combined with acute ischemic stroke (AIS) and their relationship with short-term prognosis.Methods:A total of 185 patients with H-type hypertension complicated with AIS admitted to the Linyi Central Hospital from January 2020 to December 2022 were selected and included in the AIS group. According to the National Institutes of Health Stroke Scale (NIHSS) score, there were 55 cases in the mild group (≤4 points), 86 cases in the moderate group (5-20 points), and 44 cases in the severe group (>20 points); According to the improved Rankin scale score, the patients were divided into a poor prognosis group (>2 points) and a good prognosis group (≤2 points). Another 100 healthy individuals who underwent physical examinations during the same period in our hospital were selected as the control group. Enzyme linked immunosorbent assay was used to detect serum sRAGE and NRG-1 levels in subjects. The correlation between serum sRAGE, NRG-1 levels and NIHSS score in patients with H-type hypertension complicated with AIS was analyzed through Spearman test. A multivariate logistic regression model was used to analyze the influencing factors of poor short-term prognosis in patients with H-type hypertension complicated with AIS. The receiver operating characteristic (ROC) curve analysis revealed the predictive value of serum sRAGE and NRG-1 levels for poor short-term prognosis in patients with H-type hypertension complicated with AIS.Results:The serum sRAGE level in the AIS group was higher than that in the control group, while the NRG-1 level was lower than that in the control group (all P<0.05). The serum sRAGE levels of patients in the mild, moderate, and severe groups increased sequentially, while the NRG-1 levels decreased sequentially (all P<0.05). Spearman correlation analysis showed that the NIHSS score was positively correlated with serum sRAGE levels ( rs=0.847, P<0.001) and negatively correlated with serum NRG1 levels ( rs=-0.810, P<0.001) in patients with H-type hypertension and AIS. After 90 days of follow-up, the incidence of poor prognosis in 185 patients with H-type hypertension and AIS was 37.84%(70/185). Multivariate logistic regression model analysis showed that increased age, diabetes, increased NIHSS score, increased homocysteine (Hcy) and increased sRAGE were independent risk factors for poor short-term prognosis of patients with type H hypertension combined with AIS, while increased NRG-1 was an independent protective factor (all P<0.05). ROC curve analysis showed that the combined detection of serum sRAGE and NRG-1 levels predicted a poor short-term prognosis in patients with H-type hypertension and AIS, with an area under the curve (AUC) of 0.876, which was higher than the predicted values of 0.795 and 0.791 for serum sRAGE and NRG-1 levels alone (all P<0.05). Conclusions:Elevated serum sRAGE levels and decreased NRG-1 levels are closely related to worsening of the condition and poor short-term prognosis in patients with H-type hypertension and AIS. The combined detection of serum sRAGE and NRG-1 levels has high predictive value for poor short-term prognosis in patients with H-type hypertension and AIS.

Objective:To explore the effect and mechanism of naringenin (NAR) on glucose and lipid metabolism and oxidative stress in rats with gestational diabetes mellitus (GDM) .Methods:10 normal pregnant rats were selected as the control group, and the GDM model was prepared. One-time tail iv streptozotocin (STZ, 35 mg/kg) was used to construct GDM model. The 50 rats successfully modeled were divided into model group, metformin group (Met group, 20 mg/kg), low NAR (NAR-L, 50 mg/kg), high (NAR-H, 100 mg/kg) dose groups and NAR+EX527 group (NAR 100 mg/kg+EX527 10 mg/kg), 10 rats per group. Rats in Met group and NAR low-dose and high-dose groups were given corresponding doses of drugs ig. Rats in NAR+EX527 group were given NAR ig and EX527 ip at the same time, once a day, for 28 consecutive days. The levels of fasting blood glucose (FBG) and fasting serum insulin (FINS) in rats were detected, and the homeostasis model assessment for insulin resistance (HOMA-IR) was calculated; the levels of serum triglycerides (TG), cholesterol (TC) and low density lipoprotein cholesterol (LDL-C) were detected; the level of malondialdehyde (MDA) in pancreatic tissue and the activity of superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px) were detected; the tissue lesions of pancreas and placenta were observed with HE staining; the expression of pancreatic tissue AMP-activated protein kinase (AMPK), p-AMPK, silent information regulator 1 (SIRT1), peroxisome proliferator-activated receptor γ coactivator-1α (PGC-1 α) proteins was detected with Western blotting method. Results:Compared with control, the FBG, FINS, HOMA-IR, serum TC, TG, LDL-C levels, and pancreatic tissue MDA level in the model group were significantly increased, the SOD and GSH-Px activities, pancreatic tissue p-AMPK/AMPK ratio and SIRT1 and PGC-1 α expression levels were significantly reduced ( P<0.05), and the placenta and pancreas tissues showed obvious pathological damage; Compared with the model group, the FBG, FINS, HOMA-IR, serum TC, TG, LDL-C levels, and pancreatic tissue MDA level in the Met group and the NAR-L and NAR-H groups were significantly reduced, the SOD and GSH-Px activities, pancreatic tissue p-AMPK/AMPK ratio and SIRT1 and PGC-1 α expression levels were significantly increased ( P<0.05), the placenta and pancreas tissues damage reduced; and on the basis of NAR intervention, the use of SIRT1 inhibitor EX527 could significantly reverse the protective effect of NAR on GDM rats. Conclusion:NAR may improve glucose and lipid metabolism disorder and oxidative stress response in GDM rats by activatin SIRT1/PGC-1 α pathway.

Objective To investigate the efficacy and safety of cryoablation combined with Camrelizumab monoclonal antibody in the treatment of hepatocellular carcinoma(HCC).Methods A total of 103 HCC patients who were admitted to our hospital from June 2020 to June 2023 were enrolled and randomly divided into combined treatment group with 53 patients and control group with 50 patients.The patients in the control group received percutaneous argon-helium cryoablation,and those in the combined treatment group received percutaneous argon-helium cryoablation combined with Camrelizumab monoclonal antibody.The two groups were compared in terms of short-term response,changes in T lymphocyte subsets after treatment,changes in liver function and alpha-fetoprotein(AFP)after treatment,and progression-free survival and overall survival during follow-up.The t-test was used for comparison of normally distributed continuous data between groups,and the chi-square test was used for comparison of categorical data between groups.The Kaplan-Meier method was used to plot survival curves,and the log-rank test was used for comparison of survival time between the two groups.Results The combined treatment group had significantly higher overall response rate and disease control rate than the control group(χ2=4.156 and 4.348,P=0.042 and 0.037).After treatment,the combined treatment group had significant increases in the percentages of CD3+and CD4+T lymphocytes and CD4+/CD8+ratio(P＜0.05)and a significant reduction in the percentage of CD8+T lymphocytes(P＜0.05),while the control group had no significant changes in T lymphocyte subsets after treatment(P＞0.05),and compared with the control group after treatment,the combined treatment group had significantly higher percentages of CD3+and CD4+T lymphocytes and CD4+/CD8+ratio(all P＜0.05)and a significantly lower percentage of CD8+T lymphocytes(P＜0.05).After treatment,both groups had significant reductions in the levels of alanine aminotransferase(ALT),aspartate aminotransferase(AST),and AFP(all P＜0.05)and a significant increase in the level of albumin(Alb)(P＞0.05),and compared with the control group after treatment,the combined treatment group had significantly lower levels of ALT,AST,and AFP(all P＜0.05)and a significantly higher level of Alb(P＜0.05).There were no significant differences in the incidence rates of grade Ⅲ—Ⅳ(moderate to severe)adverse reactions between the two groups(P＞0.05).Compared with the control group,the combined treatment group had significantly better median progression-free survival(21.32 months vs 15.31 months,χ2=4.689,P=0.030)and median overall survival(28.36 months vs 20.75 months,χ2=5.030,P=0.025).Conclusion Argon-helium cryoablation combined with Camrelizumab monoclonal antibody can effectively improve short-term response,enhance immune function,and prolong survival time,with a favorable safety profile.

Objective:To explore the effect and mechanism of naringenin (NAR) on glucose and lipid metabolism and oxidative stress in rats with gestational diabetes mellitus (GDM) .Methods:10 normal pregnant rats were selected as the control group, and the GDM model was prepared. One-time tail iv streptozotocin (STZ, 35 mg/kg) was used to construct GDM model. The 50 rats successfully modeled were divided into model group, metformin group (Met group, 20 mg/kg), low NAR (NAR-L, 50 mg/kg), high (NAR-H, 100 mg/kg) dose groups and NAR+EX527 group (NAR 100 mg/kg+EX527 10 mg/kg), 10 rats per group. Rats in Met group and NAR low-dose and high-dose groups were given corresponding doses of drugs ig. Rats in NAR+EX527 group were given NAR ig and EX527 ip at the same time, once a day, for 28 consecutive days. The levels of fasting blood glucose (FBG) and fasting serum insulin (FINS) in rats were detected, and the homeostasis model assessment for insulin resistance (HOMA-IR) was calculated; the levels of serum triglycerides (TG), cholesterol (TC) and low density lipoprotein cholesterol (LDL-C) were detected; the level of malondialdehyde (MDA) in pancreatic tissue and the activity of superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px) were detected; the tissue lesions of pancreas and placenta were observed with HE staining; the expression of pancreatic tissue AMP-activated protein kinase (AMPK), p-AMPK, silent information regulator 1 (SIRT1), peroxisome proliferator-activated receptor γ coactivator-1α (PGC-1 α) proteins was detected with Western blotting method. Results:Compared with control, the FBG, FINS, HOMA-IR, serum TC, TG, LDL-C levels, and pancreatic tissue MDA level in the model group were significantly increased, the SOD and GSH-Px activities, pancreatic tissue p-AMPK/AMPK ratio and SIRT1 and PGC-1 α expression levels were significantly reduced ( P<0.05), and the placenta and pancreas tissues showed obvious pathological damage; Compared with the model group, the FBG, FINS, HOMA-IR, serum TC, TG, LDL-C levels, and pancreatic tissue MDA level in the Met group and the NAR-L and NAR-H groups were significantly reduced, the SOD and GSH-Px activities, pancreatic tissue p-AMPK/AMPK ratio and SIRT1 and PGC-1 α expression levels were significantly increased ( P<0.05), the placenta and pancreas tissues damage reduced; and on the basis of NAR intervention, the use of SIRT1 inhibitor EX527 could significantly reverse the protective effect of NAR on GDM rats. Conclusion:NAR may improve glucose and lipid metabolism disorder and oxidative stress response in GDM rats by activatin SIRT1/PGC-1 α pathway.

Objective Conduct routine microbiological monitoring of laboratory rats and mice from vendors to provide an important basis for the scientific management of laboratory animal facility and ensure the reliability of relevant experimental data obtained from laboratory animals.Methods Taking the Department of Laboratory Animal Science of Fudan University as an example, between April 2021 and April 2023, rats and mice purchased from 7 vendors were sampled for microbiological quality according to the principle of simple random sampling on the arrival days of animal delivery. Then, surveillance tests were conducted to examine the microbiological contaminations according to the national standards of SPF laboratory animals.Results The total qualified rate was 80.36%, with 52.63% in SD rat, 82.76% in inbred mice, 86.67% in outbred mice and 86.36% in immunodeficient mice in details. The most frequent bacteria isolated were Staphylococcus aureus, Pseudomonas aeruginosa, Klebsilla pneumoniae and Rodentibacter heylii, and their detection rates were 10.76%, 3.16%, 2.53% and 0.63%, respectively. Serological assays demonstrated the highest prevalence for virus was Sendai virus, and the detection rate was 2.53%. In addition to the pathogens those must be excluded from SPF rodents, Entamoeba muris and Enterobacter spp. were also detected in inbred mice, and Klebsiella oxytoca was detected in immunodeficient mice, with the detection rates of 1.15%, 2.30% and 4.55%, respectively.Conclusion There are certain incidences of pathogen infections in laboratory rats and mice from vendors, and an efficient microbiological monitoring of laboratory animals should be implemented in animal facilities, in order to eliminate pathogen infections in laboratory animals, which is required for improving the accuracy of research results and protecting the occupational health of laboratory animal practitioners as well.