Objective:The contaminated microorganisms in the whole production process of Yujin,mulberry leaf and cicada were investigated,and a database was established to explore the microbial key control points and control measures in the production process of traditional Chinese medicine(TCM)formula granules,in order to provide demonstration research for enterprises,provide basis for formulating standards and improving production processes,and fill the blank of the whole.Methods:The raw materials,intermediate products and final products of three kinds of TCM formula particles were analyzed according to the Chinese Pharmacopoeia 2020,and the supporting mi-croorganisms were identified by mass spectrometry technology.The degree of exogenous polluting microorganisms from raw materials to finished products of TCM formula particles and the types of microorganisms easily contamina-ted were analyzed and studied.Results:The raw materials(TCM decoction pieces)of the three TCM formulations granules had serious microbial pollution,and the thermostable bacillus of the raw materials could still be detected in the intermediate products and final products even after the process treatment,and the total number of aerobic bacte-ria in some batches exceeded the limit.Enterococcus faecium from human and animal intestines was also detected in intermediate products and finished products.Conclusion:①Strengthening the control of microbial contamination of TCM decoction pieces is conducive to reducing the microbial load of final products;②There are risks of spore microorganism contamination in the production technology of 3 kinds of formula granuleshas,and the production process parameters should be optimized to reduce the microbial load;③Pharmaceutical companies need to strength-en the whole process of product control to reduce the risk of microbial contamination.

Objective To evaluate the feasibility,reliability and validity of the symptom identification scale for kidney yang deficiency syndrome.Methods The symptom identification scale for kidney yang deficiency syndrome developed in previous research was administered to 200 vascular cognitive impairment patients from September 2020 to September 2022 to assess feasibility through recall rate,completion rate,and completion time;reliability was measured using retest reliability,split-half reliability,homogeneity reliability,and inter-rater reliability;and validity was evaluated based on discriminant and structural validity.Results A total of 200 scales were sent out,and all of them cooperated and were completed and retrieved within 20 min.The results of reliability analysis showed that the retest reliability of the scale was 0.828 for the dimension of yang deficiency and 0.718 for the dimension of kidney qi deficiency;the Spearman-Brown coefficient of split-half reliability was 0.784;the Cronbach coefficient of the dimension of yang deficiency was 0.799,and the Cronbach coefficient of the dimension of kidney qi deficiency in the homogeneity reliability was 0.670.The results of the analysis showed that the differences between the kidney yang deficiency syndrome group and the non-kidney yang deficiency syndrome group in the yang deficiency dimension,kidney qi deficiency dimension scores and total scale scores in the discriminant validity were statistically significant(P<0.001);the KMO value in the structural validity was 0.842,and a total of two factors with eigenvalues greater than 1 were extracted,with a cumulative variance contribution rate of 58.227%.Conclusion The symptom identification scale for kidney yang deficiency syndrome demonstrates adequate reliability and validity,potentially enhancing the prediction of kidney yang deficiency in vascular cognitive impairment.However,the validity of the scale is somewhat limited and requires further refinement for clinical application.

Sirtuin2(SIRT2)is an NAD+dependent histone deacetylase that plays a key role in maintaining cellular REDOX potential and modulating pro-inflammatory immune responses.How-ever,its role in acute kidney injury(AKI)has not been proven.To explore the role of SIRT2 in AKI,AKI models were constructed in wild-type(WT)and SIRT2 knockout(SIRT2-/-)mice by injection of lipopolysaccharide(LPS).HE results showed that kidney damage in SIRT2-/-mice was more significant than that in LPS treated WT mice.qRT-PCR and Western blot results showed that more significant changes in inflammatory genes,proteins and oxidative stress proteins in SIRT2-/-mice.The results suggest that SIRT2 deficiency exacerbates LPS induced AKI.

The negative effects of low temperature can readily induce a variety of diseases. We sought to understand the reasons why cold stress induces disease by studying the mechanisms of fine-tuning in macrophages following cold exposure. We found that cold stress triggers increased macrophage activation accompanied by metabolic reprogramming of aerobic glycolysis. The discovery, by genome-wide RNA sequencing, of defective mitochondria in mice macrophages following cold exposure indicated that mitochondrial defects may contribute to this process. In addition, changes in metabolism drive the differentiation of macrophages by affecting histone modifications. Finally, we showed that histone acetylation and lactylation are modulators of macrophage differentiation following cold exposure. Collectively, metabolism-related epigenetic modifications are essential for the differentiation of macrophages in cold-stressed mice, and the regulation of metabolism may be crucial for alleviating the harm induced by cold stress.
Acetylation ; Animals ; Cold-Shock Response ; Epigenesis, Genetic ; Macrophages/metabolism* ; Mice ; Mitochondria/metabolism*

Objective:To investigate the regulatory mechanism and clinical significance of RASSF5 in cutaneous melanoma.Methods:We used GEPIA and HPA online tools and TCGA and GSE53118 data sets for differential expression analysis and prognosis analysis; cBioPortal was applied to analyze the correlation between RASSF5 expression and gene mutation and methylation; "limma" R package was used to screen differential genes for GO analysis, KEGG analysis and protein-protein interaction analysis, and we also performed GSEA analysis on all genes; finally we used ESTIMATE and CIBERSORTx online tools to evaluate immune cell infiltration. The kruskal test was used to compare the differences between the groups for measurement data, the Kaplan-Meier method was used to draw the survival curve, and the Cox proportional hazard regression model was used for univariate and multivariate analysis.Results:This study found that in cutaneous melanoma, the expression level of RASSF5 was significantly lower than that of normal tissues, and the methylation level was negatively correlated. KEGG analysis and GSEA analysis showed that RASSF5 can be related to multiple signaling pathways such as immune regulation, KRAS, and P53. Further analysis showed that the expression level of RASSF5 was positively correlated with the degree of infiltration of a variety of tumor immune cells. Survival analysis found that the expression level of RASSF5 was correlated with the overall survival rate of patients with cutaneous melanoma, and multivariate regression analysis found that RASSF5 was an independent predictor of cutaneous melanoma. Finally, using the GSE53118 dataset to verify again, RASSF5 is related to the overall survival rate of patients with cutaneous melanoma and can be used as an independent predictor.Conclusions:RASSF5 may regulate the occurrence and development of cutaneous melanoma cells through a variety of ways. It is a potential biological prognostic marker and therapeutic target.

Objective:To investigate the regulatory mechanism and clinical significance of RASSF5 in cutaneous melanoma.Methods:We used GEPIA and HPA online tools and TCGA and GSE53118 data sets for differential expression analysis and prognosis analysis; cBioPortal was applied to analyze the correlation between RASSF5 expression and gene mutation and methylation; "limma" R package was used to screen differential genes for GO analysis, KEGG analysis and protein-protein interaction analysis, and we also performed GSEA analysis on all genes; finally we used ESTIMATE and CIBERSORTx online tools to evaluate immune cell infiltration. The kruskal test was used to compare the differences between the groups for measurement data, the Kaplan-Meier method was used to draw the survival curve, and the Cox proportional hazard regression model was used for univariate and multivariate analysis.Results:This study found that in cutaneous melanoma, the expression level of RASSF5 was significantly lower than that of normal tissues, and the methylation level was negatively correlated. KEGG analysis and GSEA analysis showed that RASSF5 can be related to multiple signaling pathways such as immune regulation, KRAS, and P53. Further analysis showed that the expression level of RASSF5 was positively correlated with the degree of infiltration of a variety of tumor immune cells. Survival analysis found that the expression level of RASSF5 was correlated with the overall survival rate of patients with cutaneous melanoma, and multivariate regression analysis found that RASSF5 was an independent predictor of cutaneous melanoma. Finally, using the GSE53118 dataset to verify again, RASSF5 is related to the overall survival rate of patients with cutaneous melanoma and can be used as an independent predictor.Conclusions:RASSF5 may regulate the occurrence and development of cutaneous melanoma cells through a variety of ways. It is a potential biological prognostic marker and therapeutic target.

Objective:To further understand the clinical features, treatment efficacy and risk factors for poor prognosis in infantile-onset renal tumors.Methods:Clinical data of 45 cases of infantile-onset renal tumors from June 2011 to November 2019 in Peking University First Hospital, Beijing Children′s Hospital, Beijing Tongren Hospital and Beijing Shijitan Hospital were analyzed retrospectively. The clinical features were summarized and the prognoses were evaluated. Multi-disciplinary diagnosis and treatment was used, including surgery, chemotherapy and radiotherapy. Kaplan-Meier analysis was used to calculate the overall survival rate and the event-free survival rate, while the chi-square test was used to analyze the risk factors for poor prognosis.Results:Among 45 patients, 24 were males and 21 females. The age of onset was 7 (ranged 3-11) months, and the length of tumor at initial diagnosis was 9.7 (ranged 4.9-25.0)cm. The International Society of Pediatric Oncology (SIOP) staging: 5 cases (11%) were in stage Ⅰ, 22 cases in stage Ⅱ (49%), 8 cases in stage Ⅲ (18%), 6 cases in stage Ⅳ (13%), and 4 cases in stage Ⅴ (9%). Risk groups included 5 cases (11%) in the low-risk group, 22 cases (49%) in the intermediate-risk group, and 18 cases (40%) in the high-risk group. Forty-four cases (98%) did not receive preoperative biopsy, 26 cases (58%) received preoperative chemotherapy, 39 cases (87%) received postoperative chemotherapy, and 2 cases (4%) received three-dimensional conformal radiotherapy. The 5-year overall survival rate was (83±7)%, and the 5-year event-free survival rate was (76±8)%. Hematuria as the first symptom (3/8 vs. 83% (30/36), χ2=7.005, P=0.024), tumor long diamete r≤8 cm (5/11 vs. 85% (28/33), χ2=5.606, P=0.027) and high-risk pathological group (7/18 vs.100% (26/26), χ2=21.928, P<0.01) were risk factors for poor prognosis of children with renal tumors in this group. Conclusion:The prognosis of children with infantile-onset renal tumors is fairly well, nevertheless the prognosis is poor in patients with hematuria as the first symptom and in high-risk pathological group.

OBJECTIVETo investigate the correlation of apolipoprotein AI (ApoAI), ApoB, ApoB/ApoAI and the severity of brain white matter lesions (WML).

METHODSA total of 648 patients with WML confirmed by brain magnetic resonance imaging (MRI) were divided into mild WML group (=386) and moderate to severe WML group (=262) according to evaluations with the Fazekas scale. The demographic data, blood biochemical parameters and the levels of ApoAI, ApoB and ApoB/AI ratio were compared between the two groups to identify the risk factors of moderate to severe WML.

RESULTSUnivariate analysis showed that age, gender, hypertension, diabetes, coronary heart disease, previous stroke, homocysteine, HDL-C, ApoAI, and ApoB/AI ratio all differed significantly between the two groups ( < 0.05), but ApoB levels were similar between them ( > 0.05). Multivariate logistic regression analysis revealed that with ApoAI and ApoB/AI ratio as the continuous variables, after adjustment for the compounding factors, ApoB/AI ratio was an independent risk factor (OR=11.456, 95% : 3.622-36.229, < 0.001) and ApoAI was an independent protective factor for moderate to severe WML (OR=0.068, 95% : 0.018-0.262, < 0.001). With the upper quartiles of ApoAI level (1.38 g/L) and ApoB/AI ratio (0.58) as their respective cutoff values, patients with a high ApoAI level and a low ApoB/AI ratio were found to have the lowest incidence of moderate to severe WML ( < 0.001).

CONCLUSIONSAn increased ApoB/AI ratio is an independent risk factor and an increased ApoAI level is an independent protective factor for moderate to severe WML.

Objective To summarize the clinical data of 194 patients with congenital hyperinsulinism(CHI),in order to provide the theoretical basis for the diagnosis,treatment and prognosis of CHI.Methods One hundred and ninety-four patients with CHI hospitalized in Beijing Children's Hospital Affiliated to Capital Medical University from February 2002 to January 2016 were recruited.Clinical data of 194 patients were collected and clinical characteristics and treatment were retrospectively analyzed.Results One hundred and forty-four cases in 194 patients were treated with Diazoxide,among whom,88 cases were responsive to Diazoxide,23 cases were unresponsive to Diazoxide,and 33 cases were uncertain to Diazoxide.Twenty-six cases in 194 cases were treated with Octreotide,among whom,16 cases were responsive to Octreotide,5 cases were unresponsive to Octreotide,and 5 cases were uncertain to Octreotide.Eleven cases in 194 cases had the 18 Fluoride-L-dihydroxyphenylalanine positron emission computerized tomography scan (18-F-L-DOPA-PET scan),among whom,7 cases were identified as focal lesion and 4 cases were identified as diffuse lesion.Twelve cases in 194 cases were treated with pancreatectomy,among whom,4 cases achieved normal blood glucose levels,4 cases still suffered from persistent hypoglycemia,3 cases had type 1 diabetes mellitus,and 1 case dropped out after the surgery.Forty-five cases in 194 cases had spontaneous remission and the remission time varied from 1 month to 8 years.Conclusions (1) Diazoxide is the first line drug of CHI.The older age onset is,the higher possibility of responsive to Diazoxide shows.(2)Octreotide is used as the second line drug.(3)18-F-L-DOPA-PET scan is the first choice to identify the location of CHI and it is useful in distinguishing between focal and diffuse forms.(4) The outcome of CHI patients after pancreatectomy is very different.A proper surgical method should be selected before the surgery.The blood glucose should be detected after surgery for a long time.(5) Some patients have the tendency of spontaneous remission.

Objective To investigate the expressions of metalloproteinase-9(MMP-9)in serum,brain and aorta matrix and tissue inhibitor of metalloproteinase-1(TIMP-1) in renovascular hypertensive rats(RHR),and to evaluate the association between blood pressure and levels of matrix metalloproteinase-9(MMP-9) and tissue inhibitor of metalloproteinase-1(TIMP-1).Methods Eighty healthy male SD rats were randomly divided into RHR group(n=40) and sham-operated group(n=40).Hypertension was induced by two-kidney,two-clip(2K-2C)clamps.Systolic blood pressure(SBP) was measured every 2 weeks during 12 weeks using a tail pressure meter.Stroke was confirmed by Longa's five-point scale and pathological examination.The expressions of MMP-9 and TIMP-1 in the brain and aorta tissues were detected by Western blot and immunohistochemistry.The levels of serum MMP-9 and TIMP-1 were measured by double-antibody sandwich enzyme-linked immunosorbent assay(DAS-ELISA).Results Compared with the sham-operated group,SBP stayed significantly elevated in the RHR group at 2,4,6,8,10 and 12weeks after the operation [(157±9.0) vs.(128±7.0),(176±10.0) vs.(122±6.0),(194±8.0)vs.(117±6.5),(202±12.0)vs.(124±8.0),(218±15.0) vs.(126.±8.5),and(224±20.0)vs.(129.±9.0) mm Hg,all P< 0.05].12 weeks after the surgery,the level of serum MMP-9 in the RHR group was kept significantly higher than that in the sham-operated group [(783.4±109.79)vs.(573.4±109.59) ng/mL,P<0.05],and the serum TIMP-1 level was lower in the RHR group than that in the sham-operated group[(313.02±83.9) vs.(976.19±191.1) pg/mL,P<0.05].MMP-9 expressions were significantly higher in the brain and aorta in the RHR group than that in the sham-operated group(both P<0.05),and TIMP-1 expressions were lower than that in the sham-operated group(both P<0.05).The Pearson correlation analysis showed that MMP-9 levels in serum,brain and aorta were positively correlated with systolic blood pressure(r=0.557,r=0.774 and r=0.661,all P<0.05),and TIMP-1 levels were negatively correlated with systolic blood pressure(r=-0.481,r=-0.535 and r=-0.685,all P<0.01).Conclusions Hypertension induces increased MMP-9 and decreased TIMP-1 in serum,brain and aorta in renovascular hypertensive rats.There are consistent alterations of circulating and tissue MMP-9 and TIMP-1 levels in renovascular hypertensive rats.There is a relationship between increased blood pressure and high MMP-9 and low TIMP-1 in serum and tissues.