OBJECTIVE To explore the inhibitory effect and its mechanism of asiaticoside on hypertrophic scar formation in rabbit ears based on the Wnt/ β -catenin pathway. METHODS The hypertrophic scar model in rabbit ears was established. Model rabbits were divided into the model group （normal saline）， asiaticoside low-dose group （12 mg/kg）， asiaticoside high-dose group （24 mg/kg）， and positive control group （tri amcinolone acetonide acetate 40 mg/kg）； additionally， a non-modeled control group （normal saline） was set up. There were 8 rabbits in each group. Rabbits in each group were injected with the corresponding drug solution/normal saline at the base within the scar area once daily for 28 consecutive days. After the last medication， the appearance of hypertrophic scars in rabbit ears was observed， and the hypertrophic scar index was measured； the levels of inflammatory factors [interleukin-6 （IL-6） and IL-10 ] in hypertrophic scar tissue of rabbits were determined； histopathological morphology of hypertrophic scar tissues in rabbit ears was observed， and the protein expression levels of collagen type Ⅰ （COLⅠ）， COLⅢ， Wnt4 and β -catenin in the hypertrophic scar tissues were detected. RESULTS Compared with control group， in the model group， the symptoms of hypertrophic scars were observed， the stratum corneum was thicker， and there was abnormal accumulation and disorganized arrangement of collagen. The hypertrophic scar index， IL-6 level， fibroblast density， area density of collagen fibers， mean optical density values of COLⅠ and COLⅢ， as well as the expression levels of Wnt4 and β -catenin proteins， were all significantly elevated （ P ＜0.05）， while the level of IL-10 was significantly decreased （ P ＜0.05）. Compared with the model group， the hypertrophic scars in asiaticoside low- and high-dose groups were significantly softened， with a thinned stratum corneum， and the above quantitative indicators were all significantly reversed （ P ＜0.05）. CONCLUSIONS Asiaticoside can inhibit the formation of hypertrophic scars in rabbit ears and alleviate the inflammatory response in scar tissue. Its mechanism of action may be related to the inhibition of activation of the Wnt/ β -catenin pathway.

Objective To analyze and summarize the clinical manifestations,ultrasound images,and pathological features of the rare disease fibro-adipose vascular anomaly(FAVA),in order to improve under-standing of the disease and reduce misdiagnosis rates.Methods The clinical manifestations,lesion locations,ultrasound images,and pathological findings of 37 patients diagnosed with FAVA in this hospital from January 2023 to December 2024 were analyzed.Results FAVA predominantly occurred in children and young and middle-aged women,mainly presenting as hard masses in the calves,progressive pain,with some cases ac-companied by movement disorders.It was rarely seen in the thighs,forearms,upper arms,and elbow joints.The disease primarily affected muscles and rarely affected nerves.When muscles and nerves were involved,it exhibited characteristic ultrasonographic features and unique imaging manifestations on magnetic resonance imaging.In immunohistochemistry,besides vascular and lymphatic markers such as CD31(vascular+),CD34(vascular+),D2-40(lymphatic+),and ERG(vascular endothelial cells+),some lesions also showed molecu-lar markers including Factor Ⅷ(+),Ki-67(<1%+),Vimentin(+),β-catenin(partial+),and SMA(partial+).Conclusion Awareness of the ultrasonographic and pathological characteristics of FAVA should be increased to reduce the misdiagnosis rate.

Objective:To establish a digital reverse transcription polymerase chain reaction（dRT-PCR）detection method for hepatitis E virus（HEV）using nanoplates，and to provide technical reference for HEV monitoring in shellfish by combining virus enrichment pretreatment methods.Methods:The annealing temperature，primer and probe concentrations of HEV dRT-PCR were optimized，and the specificity of the method was evaluated；the sensitivity of this method for detecting HEV in water samples and oyster extracts was compared. The inhibition rate and recovery rate of HEV detection in artificially contaminated oyster samples were calculated，commercially available oyster samples were tested，and compare them with real-time fluorescence quantitative RT-PCR（qRT-PCR）method.Results:The optimized annealing temperature for HEV dRT-PCR was determined to be 60 ℃，and the final concentrations of primers and probes were 0.4 μmol/L，0.4 μmol/L，and 0.2 μmol/L，respectively，indicating good specificity. The sensitivity of both methods for detecting HEV RNA in water samples was higher than that in oyster extracts. The recovery rates of HEV in oyster specimens contaminated with HEV fecal suspension by dRT-PCR and qRT-PCR were 18.76% and 18.36%，respectively，with no statistically significant difference（ P>0.05）；the inhibition rates were 17.26% and 9.58%，respectively，with statistically significant differences（ P<0.05）；55 commercially available oyster samples were tested，and both methods detected HEV RNA positivity in the same sample. Conclusion:The dRT-PCR method established in this study，combined with “proteinase K digestion，PEG/NaCl precipitation，and chloroform/n-butanol extraction” pretreatment，has a good recovery effect on HEV in shellfish food containing a large amount of PCR inhibitors，and can achieve absolute quantification. It has certain application value in monitoring and risk assessment of HEV in shellfish food.

Objective:To establish a digital reverse transcription polymerase chain reaction（dRT-PCR）detection method for hepatitis E virus（HEV）using nanoplates，and to provide technical reference for HEV monitoring in shellfish by combining virus enrichment pretreatment methods.Methods:The annealing temperature，primer and probe concentrations of HEV dRT-PCR were optimized，and the specificity of the method was evaluated；the sensitivity of this method for detecting HEV in water samples and oyster extracts was compared. The inhibition rate and recovery rate of HEV detection in artificially contaminated oyster samples were calculated，commercially available oyster samples were tested，and compare them with real-time fluorescence quantitative RT-PCR（qRT-PCR）method.Results:The optimized annealing temperature for HEV dRT-PCR was determined to be 60 ℃，and the final concentrations of primers and probes were 0.4 μmol/L，0.4 μmol/L，and 0.2 μmol/L，respectively，indicating good specificity. The sensitivity of both methods for detecting HEV RNA in water samples was higher than that in oyster extracts. The recovery rates of HEV in oyster specimens contaminated with HEV fecal suspension by dRT-PCR and qRT-PCR were 18.76% and 18.36%，respectively，with no statistically significant difference（ P>0.05）；the inhibition rates were 17.26% and 9.58%，respectively，with statistically significant differences（ P<0.05）；55 commercially available oyster samples were tested，and both methods detected HEV RNA positivity in the same sample. Conclusion:The dRT-PCR method established in this study，combined with “proteinase K digestion，PEG/NaCl precipitation，and chloroform/n-butanol extraction” pretreatment，has a good recovery effect on HEV in shellfish food containing a large amount of PCR inhibitors，and can achieve absolute quantification. It has certain application value in monitoring and risk assessment of HEV in shellfish food.

Autoimmune hepatitis (AIH) is a chronic progressive inflammatory disease of the liver caused by the attack of liver cells by the autoimmune system, with the features of positive serum autoantibodies, high IgG, and/or γ-globulinemia. Current studies on pregnancy in patients with AIH mainly focus on labor complications, and there is still a lack of systematic recommendations for the evaluation, treatment, and management of diseases in the progestational stage, during pregnancy, and after delivery. Although immunity is suppressed during pregnancy, poor disease control within one year before pregnancy and spontaneous drug withdrawal during pregnancy can significantly increase adverse pregnancy outcomes. Therefore, this article describes how to implement multidisciplinary collaboration and management of the whole cycle of pregnancy, so as to improve maternal and fetal safety.

Objective:To understand the current state of renal replacement therapy (RRT) in intensive care unit (ICU) of Tianjin public hospital, and to provide scientific evidence and direction for homogenized management and overall level improvement of RRT in Tianjin.Methods:The questionnaires were distributed to the chief or key staff of 33 ICUs from 32 public hospitals in Tianjin by clinical quality control center for critical care medicine of Tianjin and ICU of Tianjin Third Central Hospital. The RRT data of ICUs from January 2020 to December 2021 was collected, including the type and size of ICU, the number of patients undergoing RRT, reasons for initiating RRT, the RRT modes, the anticoagulation and the complications of RRT and so on.Results:A total of 33 valid questionnaires were obtained, with a recovery rate of 100%. The result showed that there were 38 803 patients admitted to the selected ICUs during investigation, and 5 456 of them (14.06%) received RRT. In most ICUs, the reasons of initiating RRT were renal failure, sepsis and volume overload. The mode of RRT was mainly continuous venovenous hemofiltration (CVVH), which was followed by continuous venovenous hemodiafiltration (CVVHDF). Carbonate replacement fluid was the first choice. Heparin was the dominant anticoagulant, and there was an increasing trend in the use of citrate anticoagulation simultaneously. However, heparin-free anticoagulation used mostly in bleeding patients. Overall, the RRT modes and anticoagulation methods were single. Thrombosis was the main iatrogenic factor interrupting RRT in most ICUs, and also the reasons for complications related to catheter or circulation pipeline. It still showed an ineffective anticoagulation of RRT even after increasing the dosage of anticoagulants.Conclusions:RRT is an important organ support method in ICU, which has been widely carried out in ICUs of Tianjin and continues to expand. Despite the positive performance, it still needs to be improved and standarized in some aspects, such as the diversification of RRT modes, anticoagulation, and the complication prevention.

Objective:To explore the cause of abnormal morphology sperm on fertilization from insemination function and provide reference for the selection of fertilization methods for teratozoospermia patients.Methods:Through a retrospective cohort study of their first in vitro fertilization (IVF) treatment cycles in Reproductive Medicine Center of Shenzhen People's Hospital from January 2016 to March 2020, all patients were divided into four groups according to the normal sperm morphology rate (NSMR), group A: IVF normal sperm morphology (NSMR≥4%, n=750), group B:mild teratozoospermia (2%≤NSMR<4%, n=277), group C: moderate teratozoospermia (1%≤NSMR<2%, n=110), group D: severe teratozoospermia (0%≤NSMR<1%, n=49). We compared normal fertilization rate, fertilization failure rate (fertilization rate<30%) and total fertilization failure rate (fertilization rate=0) among the four groups and we also compared insemination function indexs: 2 h tyrosine phosphorylation rate, hyaluronan-binding assay (HBA) positive rate, content of acrosin, spontaneous acrosome reaction rate and induced acrosome reaction rate. Results:1) The normal fertilization rate of group D [52.4%(18.3%, 69.0%)] was significantly lower than that of group A [60.0%(45.5%, 75.0%), P=0.008] and group B [60.0%(42.9%, 75.0%), P=0.028]; the fertilization failure rate [22.4% (11/49)] was significantly higher than that of group A [5.5% (41/750), P<0.001] and group B [8.3% (23/277), P=0.018]; the total fertilization failure rate [14.3% (7/49)] was significantly higher than that of group A [2.7% (20/750), P=0.006]. Multivariate logistic regression models: the normal fertilization rate of group D was significantly lower than that of group A ( OR=0.433, P=0.008), and the risk of fertilization failure ( OR=5.426, P<0.001) and total fertilization failure ( OR=8.194, P<0.001) were significantly higher than those of group A. 2) HBA positive rate in groups B, C, D [75.0%(62.3%, 83.0%), 71.0%(58.0%, 81.0%), 68.0%(48.0%, 76.5%)] was significantly lower than that in group A [80.0%(71.0%, 85.0%), all P<0.001] and induced acrosome reaction rate in group C and group D [32.3%(26.5%, 40.8%), 28.8%(24.2%, 43.0%)] was significantly lower than that in group A [37.8%(30.5%, 46.8%), P<0.001, P=0.009]. 3) Spearman correlation analysis showed that spem normal morphology rate was positively correlated with HBA positive rate ( r=0.259, P<0.001) and induced acrosome reaction rate ( r=0.202, P<0.001). 4) Receiver operating characteristic (ROC) curve analysis was performed to determine a cut-off value using HBA positive rate, induced acrosome reaction rate and sperm normal morphology rate as independent variables with the fertilization rate of IVF cycles (normal sperm morphology rate <4%) dichotomized at 30%. The best cut-off value of HBA positive rate obtained was 73.5% with a sensitivity of 51.4% and specificity of 73.8% [area under curve (AUC) (95% CI)=0.643 (0.559-0.726), P=0.002]; the cut-off value of induced acrosome reaction rate was 28.9% with a sensitivity of 72.1% and specificity of 50% [AUC (95% CI)=0.599 (0.497-0.700), P=0.036]; the cut-off value of normal sperm morphology rate was 1.45% with a sensitivity of 77.8% and specificity of 42.9% [AUC (95% CI)=0.605 (0.509-0.701), P=0.025]. Conclusion:Abnormal morphology sperm may affect IVF fertilization by HBA positive rate and induced acrosome reaction rate. For teratozoospermia patients, especially for the severe teratozoospermia (0%≤NSMR<1%), we recommend that HBA positive rate and induced acrosome reaction rate are tested after ovulation induction treatment. If the post-treatment sperm meets the requirements of routine IVF fertilization on the day of retrieved oocytes, but HBA positive rate<73.5%, induced acrosome reaction rate<28.9%, short time IVF or intracytoplasmic sperm injection (ICSI) or half-ICSI is recommended to minimize IVF fertilization failure.

Objective:To explore the cause of abnormal morphology sperm on fertilization from insemination function and provide reference for the selection of fertilization methods for teratozoospermia patients.Methods:Through a retrospective cohort study of their first in vitro fertilization (IVF) treatment cycles in Reproductive Medicine Center of Shenzhen People's Hospital from January 2016 to March 2020, all patients were divided into four groups according to the normal sperm morphology rate (NSMR), group A: IVF normal sperm morphology (NSMR≥4%, n=750), group B:mild teratozoospermia (2%≤NSMR<4%, n=277), group C: moderate teratozoospermia (1%≤NSMR<2%, n=110), group D: severe teratozoospermia (0%≤NSMR<1%, n=49). We compared normal fertilization rate, fertilization failure rate (fertilization rate<30%) and total fertilization failure rate (fertilization rate=0) among the four groups and we also compared insemination function indexs: 2 h tyrosine phosphorylation rate, hyaluronan-binding assay (HBA) positive rate, content of acrosin, spontaneous acrosome reaction rate and induced acrosome reaction rate. Results:1) The normal fertilization rate of group D [52.4%(18.3%, 69.0%)] was significantly lower than that of group A [60.0%(45.5%, 75.0%), P=0.008] and group B [60.0%(42.9%, 75.0%), P=0.028]; the fertilization failure rate [22.4% (11/49)] was significantly higher than that of group A [5.5% (41/750), P<0.001] and group B [8.3% (23/277), P=0.018]; the total fertilization failure rate [14.3% (7/49)] was significantly higher than that of group A [2.7% (20/750), P=0.006]. Multivariate logistic regression models: the normal fertilization rate of group D was significantly lower than that of group A ( OR=0.433, P=0.008), and the risk of fertilization failure ( OR=5.426, P<0.001) and total fertilization failure ( OR=8.194, P<0.001) were significantly higher than those of group A. 2) HBA positive rate in groups B, C, D [75.0%(62.3%, 83.0%), 71.0%(58.0%, 81.0%), 68.0%(48.0%, 76.5%)] was significantly lower than that in group A [80.0%(71.0%, 85.0%), all P<0.001] and induced acrosome reaction rate in group C and group D [32.3%(26.5%, 40.8%), 28.8%(24.2%, 43.0%)] was significantly lower than that in group A [37.8%(30.5%, 46.8%), P<0.001, P=0.009]. 3) Spearman correlation analysis showed that spem normal morphology rate was positively correlated with HBA positive rate ( r=0.259, P<0.001) and induced acrosome reaction rate ( r=0.202, P<0.001). 4) Receiver operating characteristic (ROC) curve analysis was performed to determine a cut-off value using HBA positive rate, induced acrosome reaction rate and sperm normal morphology rate as independent variables with the fertilization rate of IVF cycles (normal sperm morphology rate <4%) dichotomized at 30%. The best cut-off value of HBA positive rate obtained was 73.5% with a sensitivity of 51.4% and specificity of 73.8% [area under curve (AUC) (95% CI)=0.643 (0.559-0.726), P=0.002]; the cut-off value of induced acrosome reaction rate was 28.9% with a sensitivity of 72.1% and specificity of 50% [AUC (95% CI)=0.599 (0.497-0.700), P=0.036]; the cut-off value of normal sperm morphology rate was 1.45% with a sensitivity of 77.8% and specificity of 42.9% [AUC (95% CI)=0.605 (0.509-0.701), P=0.025]. Conclusion:Abnormal morphology sperm may affect IVF fertilization by HBA positive rate and induced acrosome reaction rate. For teratozoospermia patients, especially for the severe teratozoospermia (0%≤NSMR<1%), we recommend that HBA positive rate and induced acrosome reaction rate are tested after ovulation induction treatment. If the post-treatment sperm meets the requirements of routine IVF fertilization on the day of retrieved oocytes, but HBA positive rate<73.5%, induced acrosome reaction rate<28.9%, short time IVF or intracytoplasmic sperm injection (ICSI) or half-ICSI is recommended to minimize IVF fertilization failure.

Primary biliary cholangitis (PBC) is an autoimmune disease. Although PBC has the features of autoimmune disease, it has poor response to immunosuppressants and good response to the drugs participating in bile acid metabolism, such as ursodeoxycholic acid. Studies have shown that the bicarbonate secretion of biliary epithelial cells is impaired in PBC patients, and bile acid not blocked by HCO3- umbrella enters biliary epithelial cells and mediates their damage and apoptosis, leading to the expression of autoantibodies in apoptotic cells and immunologic injury. In order to explore the role of HCO3- umbrella secreted by biliary epithelial cells in the pathogenesis of PBC, this article briefly introduces the physiological function and production mechanism of HCO3- umbrella and the influencing factors for HCO3- secretion, and it is pointed out that reduced HCO3- secretion may be a key link in the pathogenesis of PBC and a potential therapeutic target.

Objective:To investigate the effect and mechanism of short-chain fatty acids (SCFAs) on the side-effect of tacrolimus on blood glucose.Methods:The C57BL/6 mice were treated with tacrolimus orally (10 mg/kg, tacrolimus group), tacrolimus plus 150 mmol/L sodium butyrate and isovalerate mixed solution (SCFAs group), broad-spectrum antibiotics (antibiotic group), and tacrolimus plus broad-spectrum antibiotics (tac&abx group). After 8 weeks intervention, the fasting blood glucose (FBG), oral glucose tolerance test (OGTT), hemoglobin A1C (HbA1c) were tested as indicators of glucose metabolism, and the gut microbiota, SCFAs concentration in the ileocecal, serum glucagon-like peptide-1 (GLP-1), fasting serum insulin, and GLP-1 expression in intestinal mucosa were performed for intestinal-glucose metabolism mechanism.Results:The FBG and HbA1c were significantly increased in tacrolimus group[(7.31±0.97)mmol/L, (8.34±1.12)%] than control group [(5.23±0.30)mmol/L, (4.32±0.80)%, all P<0.05], which remained normal in antibiotic group [(4.92±0.31)mmol/L, (5.61±0.98)%)], tac&abx group[(5.95±0.37)mmol/L, (4.56±0.26)%] and SCFAs groups [(5.87±0.68)mmol/L, (5.07±1.79)%]. The OGTT in the tacrolimus group showed glucose tolerance impairment, while other groups remained normal. The ileocecal butyric acid and isovaleric acid concentrations in the tacrolimus group were (722.3±262.2) μg/g and (10.0±5.1)μg/g, lower than the control group[ (1 321.3±165.5) μg/g, (19.7±3.6)μg/g, P<0.05]. The above acids in the SCFAs group remained normal as in the control group [(1 375.7±451.6) μg/g, (24.5±11.5)μg/g)]. The fasting serum insulin in the tacrolimus group decreased significantly to (3.2 ± 0.6)mIU/L, compared with control[ (4.4±0.9) mIU/L]and SCFAs groups [(7.0±1.1) mIU/L]. The GLP-1 test indicated a significant decrease in the tacrolimus group[ (4.7±2.9)pg/ml, P<0.05] compared with the SCFAs group and control group [(42.5±19.9) pg/ml, (33.1±9.1) pg/ml]. Conclusions:Tacrolimus affects glucose metabolism through the SCFAs-associated GLP-1 pathway in the intestine, and oral supplementation with mixed SCFAs provides a new insight for the prevention and treatment of tacrolimus-induced hyperglycemia in transplant recipients.