This paper summarized the experience of professor WANG Yongyan in differentiating and treating vascular cognitive impairment （VCI）. It is believed that the core etiology and pathogenesis of VCI are deficient qi retention and stagnation， and toxin damaging brain collaterals. The essence lies in depletion of essence and qi， and malnutrition of brain collaterals as the root cause， while phlegm， turbidity， stasis， and heat blocking brain collaterals act as the branch aspects. It reveals the pathogenesis essence of VCI as stagnation caused by deficiency and qi stagnation transforming into excess， internal generation of toxin and collateral injury. The overall therapeutic principle is to tonify deficiency， move stagnation， resolve toxin， and unblock the collaterals， guided by the three core treatment methods of nourishing， unblocking， and clearing. Clinically， according to the different characteristics of the plateau phase， fluctuation phase， and decline phase， treatment methods including replenishing qi and blood， tonifying the kidney and supplementing essence； promoting sanjiao （三焦） circulation and activating blood to dredge collaterals； clearing heat and resolving toxin， eliminating phlegm and reducing turbidity are respectively adopted. This provides clinical guidance for the treatment of VCI in traditional Chinese medicine.

Objective Currently, there is no commercially available quantitative detection kit for the main Felis domestic allergen (Fel d 1) in China. To establish a rapid detection method for Fel d 1, this study aims to prepare monoclonal antibodies against Fel d 1 protein. Methods The codon preference of Escherichia coli was utilized to optimize and synthesize the Fel d 1 gene. The prokaryotic expression plasmid pET-28a-Fel d 1 was constructed and used to express and purify the recombinant Fel d 1 protein. Subsequently, the recombinant protein was immunized into BALB/c mice and monoclonal antibodies (mAbs) were prepared by the hybridoma technique. An indirect ELISA was established using the recombinant Fel d 1 as the coating antigen, and hybridoma cell lines were screened for positive clones. The specificity and antigenic epitopes of the mAbs were confirmed by Western blot analysis. Finally, the selected hybridoma cells were injected into the peritoneal cavities of BALB/c mice for large-scale monoclonal antibody production. Results The recombinant plasmid pET-28a-Fel d 1 was successfully constructed, and soluble Fel d 1 protein was obtained after optimizing the expression conditions. Western blot and antibody titer assays confirmed the successful isolation of two hybridoma cell lines, 7D11 and 5H4, which stably secreted mAbs specific to Fel d 1. Antibody characterization revealed that the 5H4 mAb was of the IgG2a subtype and could recognize the amino acid region 105-163 of Fel d 1, while the 7D11 mAb was the IgG1 subtype and could recognize the amino acid region 1-59. Conclusion The high-purity recombinant Fel d 1 protein produced in this study provides a promising alternative for clinical immunotherapy of cat allergies. Furthermore, the monoclonal antibody prepared in this experiment lays a material foundation for the in-depth study of the biological function of Fel d 1 and the development of ELISA detection.
Animals ; Antibodies, Monoclonal/biosynthesis* ; Mice, Inbred BALB C ; Cats ; Mice ; Allergens/genetics* ; Glycoproteins/genetics* ; Enzyme-Linked Immunosorbent Assay ; Hybridomas/immunology* ; Recombinant Proteins/genetics* ; Female ; Antibody Specificity

Dent disease is a rare X-linked recessive inherited renal tubular disorder characterized by low molecular weight proteinuria (LMWP), hypercalciuria, nephrocalcinosis, and other clinical features, and can lead to progressive renal failure. It is primarily caused by mutations in the CLCN5 gene. This article reports the case of a 10-year-old male patient of Chinese descent who was incidentally found to have asymptomatic proteinuria during a routine health examination. Comprehensive biochemical testing and clinical evaluation revealed significant LMWP and hypercalciuria, while renal biopsy showed mesangial cell and matrix proliferation. Whole exome sequencing identified a novel deletion mutation in the CLCN5 gene (NM_001127899.4, c.1158delC, p.F387Lfs*42) causing a frameshift and premature termination, which is likely to disrupt its role in chloride/hydrogen ion exchange and endosomal acidification. Bioinformatic analysis indicated the variant is pathogenic. Genetic testing plays an important role in diagnosing rare kidney diseases. Early identification of pathogenic mutations is essential for facilitating timely intervention and appropriate management, potentially enhancing patient outcomes. This report expands the CLCN5 mutation spectrum and contributes to understanding the genetic and molecular mechanisms of Dent disease.
Humans ; Chloride Channels/genetics* ; Dent Disease/genetics* ; Male ; Child ; Computational Biology ; Mutation ; Proteinuria/genetics* ; Hypercalciuria/genetics*

Hepatitis B virus (HBV) infection is a major global public health problem. There will be about 257 million chronic HBV-infected patients worldwide, according to the World Health Organization's estimation by 2025. Currently, the main drugs for the treatment of chronic HBV infection are nucleos(t)ide analogues and pegylated interferon (PEG-IFN). However, previous research results show that whether it is nucleos(t)ide analogues and PEG-IFN-α monotherapy or combination therapy, or sequential combination therapy, the rate of hepatitis B surface antigen seroconversion in patients is low, and there is still a high risk of disease progression after a certain course of antiviral treatment. Therefore, to achieve the ambitious target of "eliminating viral hepatitis by 2030" set by the World Health Organization and help more hepatitis B patients achieve functional cure, a large number of new drugs have been developed and entered clinical trials. This paper summarizes and reviews the types, safety, and efficacy of new drugs to further promote advancements in the field of treatment of chronic HBV infection.

Objective:To explore the effects and possible mechanism of intestinal microbiota on lung injury in mice with acute necrotizing pancreatitis (ANP).Methods:The experimental mice were randomly assigned to normal control group (CON group), ANP model group (ANP group) and intestinal germ-free group (ABX group), with 6 mice in each group. The ANP mouse model was constructed by intraperitoneal injection of caerulein (100 μg/kg, for 10 times) at an interval of 1 hour each time, followed by 10 mg/kg lipopolysaccharide injection. Mice in ABX group were treated by Abx solution (0.5 g/L vancomycin, 1 g/L neomycin, 1 g/L metronidazole, and 1 g/L ampicillin), 1 ml/100 g gavage for 28 days before preparation of the ANP model. The CON group was injected intraperitoneally with an equal volume of PBS. Histopathologic examination of the pancreas, lungs, and terminal ileum was routinely performed. Serum amylase levels were measured using enzymatic kinetic chemistry, and serum diamine oxidase (DAO) and lung tissue myeloperoxidase (MPO) activities were measured using ELISA assay. Expression of inflammatory factors, pyroptosis-related molecules in lung tissue and intestinal epithelial tight junction proteins was detected by fluorescence quantitative PCR. Western blotting was used to detect the expression of the pyroptosis molecules caspase-1 and GSDMD in lung tissue, and intestinal epithelial tight junction proteins. Changes of bacterial distribution in lung tissue were measured by fluorescence in situ hybridization. Results:The pathological scores of pancreatic tissue of CON, ANP, and ABX group were (0.67±0.26), (7.33±0.82), and (5.67±0.81); the pathological scores of lung tissue were (1.67±0.41), (5.67±0.41), and (3.58±0.58); the pathological scores of ileal tissue were (0.58±0.52), (3.83±0.75), and (4.33±0.82); the serum amylase levels were (403.95±93.11), (1037.24±126.77), and (647.32±145.90)U/L; the MPO levels in lung tissue were (0.23±0.03), (0.63±0.09), and (0.48±0.05)U/g. ABX group had significantly lower scores in pancreatic and lung tissues, serum amylase levels, and MPO levels in lung tissue compared to ANP group, and all the differences were statistically significant (all P value <0.05). The expression level in pancreatis tissue from CON, ANP and ABX group of IL-1β mRNA was 1.84±0.90, 36.26±5.56 and 16.65±6.43, IL-6 mRNA was 1.07±0.15, 2.90±0.42 and 1.34±0.62, TNF-α mRNA was 0.47±0.11, 0.76±0.11 and 0.46±0.07, HMGB1 mRNA was 0.38±0.02, 0.72±0.22 and 0.44±0.08, caspase-1 mRNA was 1.07±0.18, 2.04±0.31 and 0.85±0.54, ASC mRNA was 1.24±0.19, 5.68±0.41 and 3.89±1.47, GSDMD mRNA was 0.79±0.17, 0.94±0.14 and 0.61±0.08, IL-18 mRNA was 0.83±0.27, 4.17±0.79 and 3.57±0.03, respectively. The expression of IL-1β, IL-6, TNF-α, HMGB1, caspase-1, ASC, and IL-18 mRNA in lung tissue was significantly increased in ANP group compared to the CON group; conversely, ABX group showed a significant decrease in the expression of these markers compared to ANP group; and all the differences were statistically significant (all P values <0.05). The protein level of caspase-1 in lung tissue of CON, ANP and ABX group was 1.59±0.51, 2.28±0.13, 1.38±0.47, and that of GSDMD was 1.90±0.09, 2.20±0.07 and 1.76±0.27, respectively, which in ANP group were significantly higher than in CON group, but in ABX group was significantly lower than in ANP group, and all the differences were statistically significant (all P values <0.05). The serum DAO levels of CON, ANP, and ABX group were (0.06±0.15), (0.52±0.11) and (0.58±0.11) ng/ml; the expression level of ileum tissue of claudin1 mRNA and protein was 0.98±0.26, 0.42±0.18, 0.32±0.24 and 1.05±0.08, 0.82±0.09, 0.19±0.04; occludin mRNA and protein was 0.91±0.07, 0.31±0.05, 0.32±0.14 and 1.03±0.07, 0.61±0.04, 0.64±0.11; ZO-1 mRNA and protein was 1.01±0.08, 0.80±0.28, 0.60±0.28, and 0.86±0.10, 0.99±0.30, 0.62±0.30. The serum DAO level was significantly elevated in both ANP and ABX groups compared to the CON group. The mRNA and protein expression of claudin-1 and occludin in both ANP and ABX groups were significantly lower than those in CON group; the expression of claudin-1 in ABX group was significantly downregulated compared to ANP group; and all the differences were statistically significant (all P values <0.05). The relative fluorescence intensities of lung tissue in CON, ANP, and ABX groups were 0.03±0.01, 0.06±0.01, and 0.04±0.01, respectively, which in ANP group was significantly higher compared to CON group, but in ABX group was significantly lower than ANP group; all the differences were statistically significant (all P values <0.05). Conclusions:Intestinal microbiota may attenuate acute pancreatitis-associated acute lung injury by inhibiting the pyroptosis pathway in lung tissue.

Objective Exploring the application effect of fragmented resistance exercise in the exercise rehabili-tation of elderly patients with chronic obstructive pulmonary disease,providing references for improving their quality of life.Methods From October to December 2024,96 elderly patients with chronic obstructive pulmonary disease admitted to a tertiary hospital in Pingdingshan city were conveniently selected as the research subjects.They were randomly divided into an experimental group and a control group(all n=48)using a random number table method.The experimental group received a fragmented resistance exercise program on the basis of routine pulmonary reha-bilitation,while the control group received routine pulmonary rehabilitation nursing care for a duration of 3 months.The lung function,exercise endurance,quality of life,and exercise compliance were compared between the 2 groups before and after intervention.Results The experimental group ultimately included 46 cases,while the control group ultimately included 47 cases.The lung function of the experimental group was better than that of the control group after 3 months of intervention(P＜0.05),and the exercise compliance scores of the experimental group were high-er than those of the control group after 1 and 3 months of intervention(P＜0.001).Repeated measures ANOVA showed that there was an interaction effect(P＜0.05)in the comparison of exercise endurance and quality of life between the 2 groups.Simple effects analysis showed that the experimental group had better exercise endurance after 3 months of intervention than the control group(P＜0.05),and better quality of life after 1 and 3 months of intervention than the control group(P＜0.05).Conclusion Fragmented resistance exercise can effectively improve lung function,enhance exercise endurance and compliance,and improve quality of life in elderly patients with chronic obstructive pul-monary disease.

Objective:To observe the pregnancy outcomes of patients with unexplained recurrent spontaneous abortion (URSA) after interventional treatment or expectant treatment.Methods:This prospective study followed up 398 patients with recurrent spontaneous abortion from March 2017 to September 2022 in seven hospitals. Among them, 267 patients were diagnosed with URSA, including 124 patients who were initially diagnosed in the interventional treatment hospital and 143 patients who were initially diagnosed in the expectant treatment hospital. All URSA patients were followed up for 33 months. Ongoing pregnancy rates were observed as main outcome indicators.Results:A total of 127 patients became pregnant, and 107 of them had sustained pregnancies, the ongoing pregnancy rate was 84.25% (107/127). The ongoing pregnancy rate was 86.11% (31/36) in the interventional treatment group and 83.52% (76/91) in the expectant treatment group, with no significant difference ( P>0.05). During the follow-up, the ongoing pregnancy rates in the interventional treatment hospital and the expectant treatment hospital were 75.71% (53/70) and 94.74% (54/57), respectively, with a significant difference ( P<0.05). The ongoing pregnancy rate after interventional treatment in the interventional treatment hospital was 82.76% (24/29), which was similar to the 94.00% (47/50) after expectant treatment in the expectant treatment hospital ( P>0.05). Conclusion:The ongoing pregnancy rate of interventional treatment for URSA patients has not been significantly improved, suggesting that it may not be necessary to carry out this treatment.

Objective:To evaluate the feasibility of training physicians from secondary comprehensive hospitals in the clinical assessment of brain death and to provide recommendations for nationwide implementation.Methods:This prospective cohort study enrolled physicians who completed standardized training in clinical brain death determination at five pilot hospitals between June and December 2023. Participants were from internal medicine, neurology, critical care, emergency, or anesthesiology departments of secondary comprehensive hospitals and had ≥5 years of clinical experience. Organ donation coordinators and surgeons involved in organ donation or transplantation were excluded. The training program comprised four modules: didactic lectures, bedside demonstrations, simulation-based practice, and written theoretical assessment with review. The theoretical assessment was considered qualified if the score was 60 or above. Participants were categorized into ≥80 and <80 groups based on assessment scores. Between-group comparisons were conducted using rank-sum or chi-square tests.Results:A total of 191 physicians from 74 secondary comprehensive hospitals were enrolled. Most held a bachelor's degree [89.5%(171/191)] and had intermediate [47.1%(90/191)] or associate senior [36.1%(69/191)] professional titles; [59.7%(114/191)] were from non-neurology specialties. The overall pass rate was 99.5% (190/191), with a mean score of 82.4±7.1. Compared with those scoring<80 (56 participants), physicians scoring ≥80 (135 participants) differed significantly by professional title, province, and department ( P=0.014, 0.019 and 0.039). The proportion scoring<80 was higher among junior/intermediate versus senior titles [38.0%(41/108) vs 18.1%(15/83), P=0.003), and among non-neurology/critical care departments (emergency, internal medicine, anesthesiology) versus neurology/critical care [39.7%(31/78) vs 22.1%(25/113), P=0.009]. Only 2.09%(4/191) achieved a perfect score. Across all test items, the overall error rate was 14.99%(700/4 670). The five knowledge points with the highest error rates were mistriggering of mechanical ventilation [96.97%(32/33)], corneal reflex [42.25%(30/71)], spinal reflexes [24.25%(65/268)], documentation of the determination [21.21%(7/33)], and the apnea test procedure [20.73%(57/275)]. Conclusions:The pilot hospitals can effectively deliver clinical training for brain death determination, supporting nationwide promotion. However, physicians' theoretical grounding in neurology at secondary comprehensive hospitals appears relatively weak. Training curricula should be optimized to further improve training quality.

Objective:To develop a Knowledge-Attitude-Practice (KAP) Scale for Dietary Management During Hemodialysis and to test its reliability and validity.Methods:Based on the KAP theoretical framework, an initial version of the scale was developed through a literature review and expert consultations. A convenience sampling method was used to recruit hemodialysis patients from four hospitals in Suzhou in March 2024. Questionnaire item analysis and reliability and validity tests were conducted.Results:A total of 460 questionnaires were distributed and 438 valid responses were collected, with an effective response rate of 95.22%. The final scale included three dimensions (knowledge, attitude, and practice) with 34 items. Content validity at the scale level was 0.910, and the item level ranged from 0.800 to 1.000. Exploratory factor analysis extracted three common factors, with a cumulative variance contribution rate of 74.520%. Confirmatory factor analysis showed a good model fit. The total Cronbach's α coefficient of the scale was 0.971, and the Cronbach's αcoefficients for the three dimensions were 0.963, 0.933, and 0.934, respectively. The test-retest reliability coefficient was 0.839.Conclusions:The Knowledge-Attitude-Practice Scale for Dietary Management During Hemodialysis demonstrates good reliability and validity, making it a valuable tool for assessing the KAP level of dietary management in hemodialysis patients.

Objective:To observe the pregnancy outcomes of patients with unexplained recurrent spontaneous abortion (URSA) after interventional treatment or expectant treatment.Methods:This prospective study followed up 398 patients with recurrent spontaneous abortion from March 2017 to September 2022 in seven hospitals. Among them, 267 patients were diagnosed with URSA, including 124 patients who were initially diagnosed in the interventional treatment hospital and 143 patients who were initially diagnosed in the expectant treatment hospital. All URSA patients were followed up for 33 months. Ongoing pregnancy rates were observed as main outcome indicators.Results:A total of 127 patients became pregnant, and 107 of them had sustained pregnancies, the ongoing pregnancy rate was 84.25% (107/127). The ongoing pregnancy rate was 86.11% (31/36) in the interventional treatment group and 83.52% (76/91) in the expectant treatment group, with no significant difference ( P>0.05). During the follow-up, the ongoing pregnancy rates in the interventional treatment hospital and the expectant treatment hospital were 75.71% (53/70) and 94.74% (54/57), respectively, with a significant difference ( P<0.05). The ongoing pregnancy rate after interventional treatment in the interventional treatment hospital was 82.76% (24/29), which was similar to the 94.00% (47/50) after expectant treatment in the expectant treatment hospital ( P>0.05). Conclusion:The ongoing pregnancy rate of interventional treatment for URSA patients has not been significantly improved, suggesting that it may not be necessary to carry out this treatment.