Objective To study the relationship between the severity of Mycoplasma pneumoniae pneumonia(MPP) and interleukin(IL)-6,IL-10,tumor necrosis factor(TNF)-α level.Methods From Jan.to Jul.2012,according to clinical pulmonary infection score(CPIS),30 children with severe MPP(severe MPP group) and 32 children with mild MPP(mild MPP group) were enrolled,and 30 healthy children were enrolled as control group).Subjects of all groups were detected for the levels of IL-6,IL-10,TNF-α,IgA,IgM and IgG.Besides,children with MPP were also detected for the levels of IL-6,IL-10 and TNF-α after treatment.Results The levels of IL-6,IL-10 and TNF-α in bronchoalveolar lavage fluid(BALF) and serum in severe MPP group were higher than those in mild MPP group and control group(P<0.05).The levels of IL-6,IL-10 and TNF-α in BALF and serum were significantly higher in mild MPP group than those in control group(P<0.05).In all children,the levels of IL-6,IL-10 and TNF-α in BALF were higher than those in serum.After treatment,the levels of IL-6 and TNF-α in BALF in children with severe MPP were significantly decreased,while that of IL-10 were increased(P<0.05).Compared with control group,the IgM level in children with MPP was increased significantly,and the IgG level was decreased notablely(P<0.05).Conclusion There might be significant correlation between levels of cytokines IL-6,IL-10,TNF-α and the disease severity of MPP in children patients,which could be used as the basis for the clinical diagnosis and treatment.

50 adult male SD rats were divided randomly into 5 groups:A,B,C1 ,C2,C3(n =1 0).The rats in roup A was used as a blank controls.The rats in group B and C were with LPS induced periodontitis,those in group C1 ,C2 and C3 received 0.2 ml of baicalin daily injection(0.01 ,0.1 and 1 .0 μg/ml respectively)into the gingival sulcus of the teeth with periodontitis for 3 days.The rats were sacri-ficed 7 days after treatment and periodontal tissues of the related teeth were observed by histology.In group C1 ,C2 and C3 the periodontal inflammation was significantly slighter than that in group B,the osteoclasts count was as following:B >C1 >C2 >C3 >A(P <0.05).The study suggests that baicalin can inhibit destructive effect of LPS to periodontal tissue of rats.

BACKGROUND:Bone marrow mesenchymal stem cells are rare in vivo. It is important to purify, proliferate and differentiate bone marrow mesenchymal stem cells in vitro for further research. OBJECTIVE:To evaluate the biological characteristics, phenotype and multiple differentiation potential cultivation of bone marrow mesenchymal stem cells that are isolated, cultured and purified using the whole bone marrow adherence method. METHODS:Bone marrow mesenchymal stem cells were isolated, purified and cultured by the whole bone marrow adherence method. Morphological observation and flow cytometry determination of cellsurface markers were performed. Osteogenic and adipogenic differentiation of bone marrow mesenchymal stem cells was induced. RESULTS AND CONCLUSION:We successful y purified and proliferated bone marrow mesenchymal stem cells with high cellviability and differentiation ability. Fibroblast-like cells were harvested, expressing CD29 and CD90, but not CD45. Fol owing osteogenic and adipogenic induction, cells were positive for oil red O staining and alizarin red staining. The whole bone marrow adherence method is easy to operate, has little impact on cellviability, and can be used to harvest high-purification bone marrow mesenchymal stem cells with high cellviability and differentiation ability.

Objective To investigate clinical value of high sensitive-cardiac troponin T(hs-cTnT) combined with creatine kinase isoenzyme MB(CK-MB) in the diagnosis of children with myocarditis.Methods From Nov.2014 to Nov.2015,a total of 102 cases of myocarditis,suspected with myocardial damage and without myocardial damage(pneumonia and capillary bronchitis),and 50 healthy children were enrolled.Plasma levels of hs-cTnT and CK-MB were detected and compared.Results The levels of plasma hs-cTnT and CK-MB in children with myocarditis were significantly higher than those without myocarditis and healthy subjects(P<0.05).Hs-cTnT and CK-MB levels in children with myocarditis,less than one month old,were significantly higher than those with age of 1 month to 3 years old(P<0.05).Conclusion Combined detection of hs-cTnT and CK-MB could be with high sensitive and specificity in diagnosis of children with myocarditis,accurately assess the disease condition and improve the therapeutic effect and prognosis,which might be worthy of clinical application.

Objective To investigate the prevalence and epidemiologic characteristics of hypothyroidism among community population in Jiangsu province. Methods The residents who had lived for at least five years and aged more than 20 years old were sampled from six layers in urban and rural districts of Jiangsu province by a stratified cluster sampling technique. Serum was sampled from 7 122 subjects and sTSH was measured by chemiluminescence immunoassay, and FT3 and FT4 were determined in the subjects with abnormal sTSH level. Results (1) The crude prevalences of overt hypothyroidism and subclinical hypothyroidism were 0.66% and 7.53% respectively in total population, with the respective standardized rates of 0.43% and 6.28%. (2)The prevalences of hypothyroidism and subclinical hypothyroidism were significantly higher in females than in males (both P <0.05). (3) The prevalence of subclinical hypothyroidism was significantly increased with advancing age in both female and males (P<0.05). Conclusion Comparing with hypothyroidism, subclinical hypothyroidism shows higher prevalence in Jiangsu province, and more attention should be paid to the follow-up and diagnosis of subclinical hypothyroidism.

Background:Early diagnosis and treatment can effectively improve the prognosis of colon cancer.Simple,effective and sensitive screening indicators are of great significance for identification of early cancer and precancerous lesions.L-selectin is a cell adhesion molecule,and podocalyxin(PODXL)is its ligand,both of them play key roles in the development of cancer.Aims:To investigate the expression and significance of L-selectin and its ligand PODXL in colon cancer.Methods:A total of 120 cases of pathological specimens(40 hyperplastic polyp,40 colon adenoma,and 40 colon cancer)from Nov.2020 to Nov.2022 at the Frist People's Hospital of Hangzhou Lin'an District and the First Affiliated Hospital of Zhejiang Chinese Medical University were collected,and 20 cases of normal intestinal mucosal tissue were served as controls.qRT-PCR and immunohistochemistry were used to detect the mRNA and protein expressions of L-selectin and PODXL,respectively.Western blotting was used to determine the expressions of L-selectin and PODXL,and their relations with different clinicopathological parameters of colon cancer were analyzed.In addition,60 serum specimens of colon cancer were collected.ELISA was used to detect serum concentrations of L-selectin and PODXL.Results:Expressions of L-selectin and PODXL mRNA and protein in colon adenoma group were significantly higher than those in normal controls and hyperplastic polyp group(P<0.05),and mRNA and protein expressions of L-selectin and PODXL in colon cancer group were significantly higher than those in normal controls,hyperplastic polyp group and colon adenoma group(P<0.05).Significant differences in protein expressions of L-selectin and PODXL were found in different pathological types,lymph node metastasis,Dukes staging in colon cancer(P<0.05).Expression of L-selectin was positive correlated with expression of PODXL in colon cancer(r=0.855,P<0.001).Serum concentrations of L-selectin and PODXL were significantly lower in the initial group than in the relapse group(P<0.05),and serum concentrations of L-selection and PODXL was significantly lower in the non-metastatic group than in the metastatic group(P<0.05).Serum concentrations of L-selectin and PODXL at 3 months after surgery was significantly lower than 3 days after surgery and before surgery(P<0.05).Conclusions:L-selectin and PODXL may be involved in the development and progression of colon cancer.They are carcinogenic proteins,and their detection could provide reference value for the prevention and early diagnosis of colon cancer,and through early screening of lesion could improve the prognosis of colon cancer to a certain extent.

Background:Studies have confirmed that colorectal tumors may be caused by normal mucosa near or inside the diverticulum,and patients with colon diverticulum are more likely to develop colon cancer than those without diverticulum.When colorectal adenoma involves diverticulum,the inflammatory state in the diverticulum will increase the risk of adenomatous dysplasia.Therefore,adenomas involving colon diverticulum are more likely to develop malignant changes,and adenomas involving colon diverticulum need to be resected to avoid colon cancer outcomes.Improved prognosis.Aims:To explore the efficacy and safety of endoscopic mucosal resection(EMR)treatment of colorectal tumors near or involving a diverticulum.Methods:Fourteen consecutive cases of adenoma(near or involving a diverticulum)during EMR treatment from Jun.2018 to Jan.2022 were collected,and analyze their clinical characteristics and outcomes.The main outcomes were adverse events,including bleeding,perforation,and electrocoagulation syndrome,while the secondary outcome was the total resection rate,block resection rate and local recurrence rate.Results:Among the 14 patients with colon adenoma,1 case was inside the diverticulum,13 cases were near the diverticulum,the diameter of the lesion was(0.76±0.25)cm,the operation time was(19.6±5.33)min.The main tissue types were tubular adenomas,and the pathologic characteristics were low grade intraepithelial neoplasia.Among the complications,delayed hemorrhage was found in 1 case(7.1%),the patient had a history of taking antiplatelet drugs(clopidogrel),electrocoagulation syndrome was found in 1 case(7.1%),and appendiceal orifice and diverticulum were involved in this patient,with no perforation event.The whole resection rate and the complete resection rate were 100%(14/14).Within 1 year after surgery,10 patients came to the hospital for re-examination of colonoscopy,and no local recurrence was found.Conclusions:EMR treatment involving colonic diverticulum adenoma is safe and effective.However,patients with adenoma involving antiplatelet drugs and appendiceal orifice and diverticulum should be alert to postoperative complications.

Enzymatic synthesis is an important way to produce β-alanine, but the biological method is expensive generally because of the high price of substrate. In this paper, a two-step enzymatic cascade system was developed, combining L-aspartase from Escherichia coli DH5α and L-aspartate α-decarboxylase from Corynebacterium glutamicum. This system catalyzes Fumarate and ammonia to β-alanine. The optimal ratio of AspA and PanD was 1:80 (W/W), and the concentration of AspA was 10 μg/mL, at 37 ℃ and pH 7.0. When the concentration of Fumarate was 100 mmol/L, the reaction reached its equilibrium after 8 h, and the yield of β-alanine was 90 mmol/L (7 g/L). The yield of β-alanine can reach 126 mmol/L (9.8 g/L) when the concentration of Fumarate increased to 200 mmol/L. Extending reaction time, the conversion rate did not change obviously. Using this two-step enzymatic cascade system, β-alanine from cheaper substrate Fumarate can be obtained.

Objective:To investigate the protective mechanism of TAK242, a specific inhibitor of Toll-like receptor 4 (TLR4), on the liver of septic rats.Methods:Eighteen male Sprague-Dawley (SD) rats were randomly divided into three groups ( n = 6 in each group). The septic model was established by intraperitoneal injection of lipopolysaccharide (LPS) 15 mg/kg. The rats in the TAK242 intervention group received intraperitoneal injection of TAK242 (5 mg/kg) before modeling, while the rats in the septic model group and the control group were injected with the same amount of solvent [10% dimethyl sulfoxide (DMSO) + 90% corn oil]. Six hours later, the blood of abdominal aorta was collected and the levels of serum alanine aminotransferase (ALT) and aspartate aminotransferase (AST) were measured by enzyme linked immunosorbent assay (ELISA). The rats were sacrificed to obtain liver, the expression levels of TLR4, tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6), cysteinyl aspartate-specific proteinase-3 (caspase-3), nuclear factor-κB p65 (NF-κB p65) and phosphorylated NF-κB p65 (p-NF-κB p65) were detected by Western blotting. Immunohistochemical staining was used to observe NF-κB p65 protein expression in liver, and hepatocyte injury was assessed by hematoxylin-eosin (HE) staining. Results:Serum ALT and AST levels in the septic model group were significantly higher than those in the control group [ALT (μg/L): 26.639±7.814 vs. 2.847±2.150, AST (μg/L): 28.442±8.417 vs. 5.779±3.019, both P < 0.01]. The ALT and AST levels in the TAK242 intervention group were significantly lower than those in septic model group [ALT (μg/L): 7.269±3.398 vs. 26.639±7.814, AST (μg/L): 3.580±3.115 vs. 28.442±8.417, both P < 0.01]. Light microscopy showed that the hepatocytes in the septic model group were disordered, with obvious cell edema and increased inflammatory cells infiltration; the hepatocytes in the TAK242 intervention group were more neatly arranged, with significantly reduced hepatocyte edema and reduced inflammatory cells infiltration. Western blotting results showed that caspase-3 protein expression in hepatic tissue of septic model group was significantly higher than that in the control group (caspase-3/GAPDH: 0.794±0.164 vs. 0.482±0.055, P < 0.05), and caspase-3 protein expression in the TAK242 intervention group significantly decreased than that in the septic model group (caspase-3/GAPDH: 0.482±0.056 vs. 0.794±0.164, P < 0.05), which indicated that TAK242 could attenuate hepatocytes apoptosis of septic rats. The expression of IL-6, TNF-α and TLR4 protein and the ratio of p-NF-κB p65/NF-κB p65 in hepatic tissue of septic model group were significantly higher than those in control group (IL-6/GAPDH: 1.442±0.204 vs. 1.019±0.024, TNF-α/GAPDH: 1.089±0.098 vs. 0.806±0.005, TLR4/GAPDH: 1.292±0.085 vs. 0.941±0.087, p-NF-κB p65/NF-κB p65 ratio: 1.936±0.081 vs. 1.579±0.183, all P < 0.05), IL-6, TNF-α and TLR4 protein expression and p-NF-κB p65/NF-κB p65 ratio in the TAK242 intervention group were significantly lower than those in septic model group (IL-6/GAPDH: 1.035±0.042 vs. 1.442±0.204, TNF-α/GAPDH: 0.572±0.096 vs. 1.089±0.098, TLR4/GAPDH: 0.984±0.078 vs. 1.292±0.085, p-NF-κB p65/NF-κB p65 ratio: 1.484±0.255 vs. 1.936±0.081, all P < 0.05), it is suggested that LPS-induced sepsis could activate the inflammatory response mediated by TLR4/NF-κB pathway in liver, and the activation of TLR4/NF-κB pathway was inhibited by TAK242 through the TLR4 pathway, therefore, the inflammation of liver in septic rats was reduced. Immunohistochemical staining showed that the positive expression of NF-κB p65 in liver was significantly increased in the septic model group compared with the control group; the positive expression of NF-κB p65 was significantly reduced in the TAK242 intervention group compared with the septic model group, and there was almost no positive expression in the nucleus. Conclusion:TAK242 could reduce liver function injury and protect the liver by inhibition TLR4/NF-κB pathway in septic rats.

Objective:To investigate the efficacy of CT imaging features in evaluating occult peritoneal metastasis (OPM) of diffuse infiltrating gastric cancer (Borrmann Type Ⅳ).Methods:Totally 101 patients with locally advanced Borrmann type Ⅳ gastric cancer were retrospectively collected who were admitted to Peking University Cancer Hospital from March 2014 to March 2021. The patients were divided into OPM group (53 cases) and the non-OPM group (48 cases) according to the results of preoperative CT and laparoscopic exploration/peritoneal cytology examination. The pathological examination results were recorded, including the degree of histological differentiation and Lauren classification. The evaluation indicators included the tumor center position, the number of tumor-occupied portions, involved orientation, mucosal broadband sign, stratified enhancement, serosa invasion, increased density of peripheral fat tissue, and enlarged lymph nodes. The maximum thickness of the primary tumor, average CT value of the primary tumor (arterial phase, venous phase, and delayed phase), difference between venous phase and arterial phase, difference between delayed phase and venous phase, and pattern of the enhanced curve were recorded. The Mann-Whitney U or Chi-square test was used to compare the differences of pathological and CT features between two groups. The multivariate logistic regression was used to screen independent predictors and establish a nomogram. The receiver operating characteristic curve was used to evaluate the performance of the nomogram in predicting OPM, and the Hosmer-Lemeshow test was used to test the model′s goodness of fit. Results:There was statistical significance in the seven indicators between the OPM and non-OPM groups, including tumor-occupied portions of stomach, mucosal broadband sign, stratified enhancement, serosa invasion, increased density of peripheral fat tissue, the enhanced curve pattern and the degree of histological differentiation ( P<0.05). Among them, the degree of histological differentiation (OR=0.19, P=0.033), stratified enhancement (OR=7.02, P=0.005) and serosa invasion (OR=14.27, P<0.001) were independent predictors of OPM. The nomogram was established based on the three significant features. The area under the curve for predicting OPM was 0.826 (95%CI 0.745-0.908), the sensitivity was 0.566 and the specificity was 0.938. The Hosmer-Lemeshow test showed a good agreement between the OPM risk predicted by the nomogram and the actual risk ( P=0.525). Conclusions:The CT features of Borrmann type Ⅳgastric cancer complicated with OPM have specific characteristics. The diagnosis model based on the degree of histological differentiation, stratified enhancement, and serosa invasion had high efficacy in evaluating OPM.