Objective To improve the quality standard of Flos Chrysanthemum Indicum from diffrent origins by analyzing on linarin and cumambrin A qualitatively and quantitatively. Methods Qualitative analysis of linarin and cumambrin A was carried out by thin layer chromatography (TLC); Content determination of linarin and cumambrin A was using high performance liquid chromatography (HPLC) on YMC C18 column (250 mm × 4.6 mm, 5 μm); The mobile phase was a mixture of acetonitrile-0.05% phosphoric acid solution; The elution mode was gradient system (0-18 min, 25%-26% A; 18-26 min, 26%-32% A; 26-33 min, 32%-34% A; 33-35 min, 34%-40% A; 35-65 min, 40%-50% A); The flow rate was 0.8 mL/min; The detection wavelengths were 203 nm and 340 nm; The column temperature was 35 ℃. Results Linarin and cumambrin A by TLC was obvious. The concents of linarin and cumambrin A in Flos Chrysanthemum Indicum from diffrent origins were different. The concents of linarin and cumambrin A from Xinyang were the highest (6.53% and 0.81% respectively). Conclusion It is the first time to establish a method to evaluate different components in Flos Chrysanthemum Indicum by TLC and HPLC. The method is simple, accurate and reproducible, which can effectively improve the existing quality standard of Flos Chrysanthemum Indicum. The result also showed Linarin and cumambrin A could reflect the quality of Flos Chrysanthemum Indicum.

Objective To investigate the expression level of hyper sensitivity C-reactive protein (hsCRP) in children infected with different pathogens in order to enhance the application value of hsCRP .Methods One hundred and one children patients with infection from March 2010 to March 2015 were selected as the research subjects and divided into the bacterial infection group (67 ca-ses) and mycoplasma infection group (34 cases) according to the hematology and bacterial culture results .Contemporaneous 50 healthy children were selected as the control group .Venous blood was collected for detecting hsCRP ,white blood cell(WBC) and neutrophil percentage(NEUT% ) .Results The levels and abnormal rates of hsCRP and WBC in the bacterial infection group were significantly higher than the those in the mycoplasma infection group and control group (P<0 .05);NEUT% and abnormal rate in the mycoplasma infection group was higher than that in the other two groups (P<0 .05) .The levels and abnormal rates of hsCRP and WBC after treatment in the bacterial infection group were lower than those before treatment ,the difference was statistically sig-nificant(P<0 .05);the area under the ROC curve of hsCRP for diagnosing bacterial infection wa maximal ,which was 0 .68 ,the sensitivity and specificity were 89 .9% and 88 .3% respectively .Conclusion The hsCRP level detection has early diagnostic value for child bacterial infection and can dynamically reflects the curative effect .

Objective To investigate the mutations of CLCNKB gene in a family with classic Bartter syndrome. Methods Genetic DNA was extracted from peripheral blood leucocytes of family members.The coding exons and intron exon junctions of CLCNKB gene were amplyfied by PCR and sequenced directly.Fifty unrelated healthy subjects were selected to exclude the possibility of polymorphism. Results A heterozygous(missense)mutation(482T>G,L161R)was detected in the exon 4 of patients.The hetemzygous mutation(L161R)was found in the mother,while no mutation was found in the father of this family.L161R had not been reported and was a novel mutation when referring to literatures and human genomic database home and abroad.Conclusion A new CLCNKB gene mutation(L161R)is identified for the first time.

The experiments was conducted on the fibroblasts of human prepuce in vitro todemonstrate the distribution of cytoplasmic microfilaments with Coomassie Blue R250.It has been found that the distribution of the cytoplasmic microfilaments canbe disturbed by the specific inhibitor,Cytochalasin B.We also observed the inhi-bition by Cytochalasin B was reversible.The cytoplasmic microfilaments regainedtheir normal distribution after the Cytochalasin B was removed.This result further confirmed that the staining method of Coomassie BrilliantBlue R 250 is reliable to demonstrate microfilaments in cultured fibrablasts.

OBJECTIVETo discuss the protective effects of Xin'ganbao Capsule (XC) on early kidney injury in streptozocin (STZ)-induced diabetic rats and its mechanisms.

METHODSTwenty-four male Wistar rats were selected to establish STZ induced diabetes mellitus (DM) model. After modeling they were randomly divided into the model group,the XC group (at the daily dose of 0.5 g/kg), and the benazepril group (at the daily dose of 4 mg/kg), 8 in each group. Another 8 rats were chosen as the blank control group. Rats in the model group and the blank control group were administered with equal volume of normal saline by gastrogavage for 8 successive weeks. The blood glucose was monitored by the end of the 4th week and the 8th week. The 24 h urine protein (24 hUP), blood urea nitrogen (BUN), and serum creatinine (SCr) were detected by the end of the 8th week. The transforming growth factor-beta1 (TGF-beta1), laminin (LN), collagen IV (Col-IV) expression were detected using immunohistochemical assay. The mRNA expressions of renal TGF-beta1, tissue inhibitor of matrix metalloproteinase-1 (TIMP-1), and plasminogen activator inhibitor-1 (PAI-1) were detected using RT-PCR. The pathological changes of the renal tissue were observed by HE, PAS, and Masson stain methods.

RESULTSCompared with the blank control group, hyperglycemia, polydipsia, polyphagia, polyuria, weight loss, emaciation, dry and dim body hair, and irritability appeared in the diabetic rats. After 8 weeks the symptoms of the two medication groups were attenuated. When compared with the blank control group, the 24 hUP, SCr, blood glucose, Col-IV, LN, TGF-beta1 positive expression ratio, the levels of TGF-beta1, TIMP-1, PAI-1 mRNA, the area of glomerular (GA), extracellular matrix (ECM), and ECM/GA all increased in the model group with statistical difference (P<0.01). The pathological changes showed obvious glomerular enlargement, the capillary loop expansion, the proliferation of the mesangial cells, increased mesangial matrix, widen and thicken glomerular basement membrane (GBM), and tubular derangement. The vacuolar degeneration and shedding could be seen in partial epithelial cells. The protein cast could also be seen with infiltration of interstitial inflammatory cells. Compared with the model group, each index of the two medication groups decreased with statistical difference (P<0.01). The pathological changes were less in the two medication groups. The mesangial cells were slightly proliferated and the mesangial matrix slightly increased. The mRNA expressions of SCr and PAI-1 were lower in the XC group than in the benazepril group (P<0.05). There was no statistical difference in the other indices between the two medication groups (P > 0.05). Conclusions XC had some protective effects and anti-glomerulosclerosis effects on early kidney injury in STZ-induced diabetic rats. Its mechanisms might be associated with down-regulating the mRNA expressions of TGF-beta1, TIMP-1, PAI-1, and Col-IV, reducing ECM and urine protein.

Animals ; Benzazepines ; therapeutic use ; Collagen Type IV ; metabolism ; Diabetes Mellitus, Experimental ; drug therapy ; metabolism ; pathology ; Drugs, Chinese Herbal ; pharmacology ; therapeutic use ; Extracellular Matrix ; metabolism ; Kidney ; pathology ; Laminin ; metabolism ; Male ; Plasminogen Activator Inhibitor 1 ; metabolism ; Rats ; Rats, Wistar ; Tissue Inhibitor of Metalloproteinase-1 ; metabolism ; Transforming Growth Factor beta1 ; metabolism

ObjectiveTo investigate the clinical application of tacrulimus (TAC, FK506) in children with primary nephrotic syndrome (NS). MethodsSixty-five primary NS children received routine or decreased-dosage glucocorticosteroid according to clinical NS types after hospitalization. At the same time, TAC was given orally with the dosage of 0.1 to 0.15 mg/kg, once every 12 hours, for 6 to 24 months. And the serum concentration of TAC was monitored during the course. ResultsAfter the treatment of TAC for 1 to 2 months, 65 patients were recovered with gradually reduced urinary protein, rapidly increased serum albumin, and improvement of cholesterol and triglycerides. Total remission rate was 83.1% and onset time was 7 to 54 days. Twelve cases experienced recurrence. Increased CD4, as well as 3/3 or 3/1 TAC genotype, indicated higher remission rate. Various pathological types had different remission rates or ratio, which were as follows: minimal change nephropathy (96.4%), mesangial proliferative glomendonephritis (90.0%), membranous nephropathy (2/3), membranous proliferative glomerulonephritis (3/5), focal segmental glomerulosclerosis (4/9). The patients would recover in the course of treatment under the conditions of TAC initial dose as 0.1 to 0.15 mg /kg per 12 hours and controlled serum concentration as 5 to 10 g/L. During the treatment, 12 cases appeared gastrointestinal symptoms, mainly as anorexia, nausea and vomiting, 1 abdominal pain, 2 headache, 1 tremor, 1 paresthesia, 3 insomnia, 4 transient increased Scr, 8 slightly increased NAG, 6 increased C3 and α-2 macroglobulin. The symptoms disappeared within one week or after stopping TAC. ConclusionsTAC is effective in primary NS children, even with abnormal liver function or tuberculosis infection. TAC can also be a substitute to cyclosporine A.

Aged ; Breast Neoplasms ; pathology ; surgery ; Female ; Follow-Up Studies ; Humans ; Mastectomy ; methods ; Metaplasia ; pathology ; Papilloma, Intraductal ; pathology ; surgery ; Sebaceous Glands ; pathology

Objective To identify the specific microRNA (miRNA) that can be taken as a molecular marker for human bone marrow-derived mesenchymal stem cells (MSCs). Methods MSCs were isolated and cultured from bone marrow through density centrifugation and then were induced to differentiate into osteoblasts and chondrocytes. Samples of MSCs, osteoblasts and chondrocytes were detected by miRNA microarrays single channel fluorescence chip to determine the expression levels of miRNAs. Significance Analysis of Microarrays ( SAM, version 2.1 ) software was used to analyze the raw data to determine the miRNAs overexpressed in MSCs, which was validated in the same sample using real time reserve transcriptase polymerase chain reaction (RT-PCR). Results MSCs were successfully isolated from bone marrow and induced to differentiate into osteoblasts and chondrocytes in vitro. Microarrays showed that eight miRNAs (has-miR-424, has-miR-34a, has-miR-593, has-miR-10a, has-miR-148a,has-miR-602, mmu-miR-709 and mmu-miR-665) were overexpressed in MSCs but underexpressed in osteoblasts. Three miRNAs including has-miR-424, PREDICTED_MIR189 and mmu-miR-665, were overexpressed in MSCs but underexpressed in chondrocytes. The has-miR-424 expression in MSCs was 6.6times higher than in osteoblasts and 4.4 times higher than in chondrocytes. The results of real time RTPCR showed that the miR-424 was overexpressed in MSCs, 3.6 times higher than that in osteoblasts and 3.1 times higher than that in chondrocytes, which was coincident with the results of microarray. Conclusions The screened MSCs express more miRNAs in comparison with osteoblasts and chondrocytes,play important roles in maintaining self renewal and undifferentiation of MSCs and is a promising specific molecule marker for MSCs.

Objectives: The aim of the study was to evaluate the therapeutic effects of 6-mercaptopurine in the treatment of refractory childhood nephrotic syndrome (NS). Methods: According to the varieties of NS, 6-mercaptopurine (2 mg/kg body weight daily) combined with corticosteroid or 6-mercaptopurine (2 mg/kg body weight daily) alone after tapering of steroids were given to 28 consecutive children with primary NS in our hospital. Results: One month after the use of 6-mercaptopurine, proteinuria was decreased. The duration of improvement was 9～28 days, with mean duration of 17 days. Over-all effective rate was 85.7%. Among different varieties of NS, the best therapeutic effect was noted in steroid-dependent children; the better therapeutic effect in steroid-resistant children; and good therapeutic effect in frequently relapsing children. The effective rates were 100%, 84.6%, 81.8% respectively. All the pathological varieties of 28 children were confirmed by renal biopsy. The better therapeutic effects were noted in slight mesangial proliferative glomerulonephritis (MsPGN) and minimal change nephrotic syndrome (MCNS). The less therapeutic effect was noted in membranoproliferative glomerulonephritis (MPGN). Their therapeutic effective rates were 92.9%, 80%, 66.7% respectively. Unfortunately, drug-induced aplastic anemia was seen in 2 cases. Slight gastrointestinal reactions were present in 6 cases. There were no side reaction on the gonad. Conclusions: The great difference in the therapeutic effects is related to the different pathologic varieties of NS. With regard to the treatment of refractory NS in children, the pathological varieties should be confirmed by renal biopsy as soon as possible. Based on the renal biopsy, 6-mercaptopurine can be considered in the treatment of MsPGN and MCNS. As a result, relapses could be reduced; the duration of remission could be prolonged, and the side reactions from steroid treatment could be avoided. The use of 6-mercaptopurine for the treatment of refractory NS is one of the effective therapy.

Objective To investigated the effects of IL -18 on middle ear allergic inflammation in rat model of OME .Methods Eighteen SD rats were randomly divided into three group :group A(control group ,n=12 ears) , group B (OME model group ,n=12 ears) ,group C (IL -18 injectiion group ,n=12 ears) .The rat model of OME was established by sensitizing with ovalbumin (OVA) and later challenging in tympanic bullae .Recombinant rat IL-18 (1 μg ,+0 .2 ml saline ,) were injected in group C at 1 ,2 ,7 ,8 ,15 ,16 day .At the same time sacle ,0 .2 ml sa-line ,instead of IL -18 ,were intraperitoneal injection in group A and B .The morphologic changes of the middle ear epithelial cells and inflammatory cells infiltration were observed under light microscope .The level of IFN -γand IL-4 in tympanic lavage fluid(TLF) were determined by ELISA .Results Pathological examination showed that middle ear mucosa inflammation and eosinophil infiltration in group C were no less severe than group B .The numbers of neutrophils in group C increased significantly compraring with group B (P<0 .05) .Numbers of eosinophils in group C were slightly increased comparing with group B (P>0 .05) ,while significantly greater than that in group A (P<0 .05) .ELISA showed that the level of IFN -γ in group C was stronger than that in group B and A (both P<0 .05) .As compared to the group A ,the expression of IL -4 in group B and group C were remarkably stronger (both P<0 .05) ,no significant difference was found between group B and group C (P>0 .05) .Addtionally ,there was no significant difference in the ratio of Th1 /Th2(IFN -γ/IL -4)between group B and group C (P>0 .05) . Conclusion IL -18 acts as an immune regulatory factors ,significantly increases Th1 cytokine IFN -γ.Although to some extent alleviate the OME rat middle ear Th1 /Th2 imbalance ,there is still excessive activation of Th cells . Th1 and Th2 cells factor are excessive for the secretion disorder of the immune response status .The OME rat mid-dle ear allergic inflammation has not been fundamentally alleviated ,the underlying mechamism should be further studied .