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 .

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.

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.

BACKGROUND:Previous animal experiments have demonstrated that mandibular advancement can cause the remodeling of temporomandibular joint tissue of young SD rats. This is mainly characterized by accelerated growth rate of the condyle tissue and secondary growth of mandible. But the ultrastructural remodeling of condylar chondrocytes remains poorly understood. OBJECTIVE:To observe the histological and ultrastructural variations of reconstructed condylar cartilage of young rats under the effect of continuous mandibular advancement. METHODS:SD rats aged 4 weeks were randomly divided into control and experimental groups. Rats in the experimental group were subjected to mandibular advancement for 24 hours and sacrificed at 3, 7, 14, 21 and 30 days of intervention. Condylar cartilage samples were harvested and their histological and ultrastructural changes were observed under optical microscope and transmission electron microscope. RESULTS AND CONCLUSION:After 14 days of intervention, the thickness of condylar cartilage in the experimental group increased first and then became thin in the period of observation. The cartilage thickness variations in the postmedian condylar were significant (P < 0.01). After 7 days of intervention, the ultrastructure of condylar chondrocytes was reconstructed, including intracelular karyopyknosis, rough endoplasmic reticulum compartment sweling, smaler even absent lipid droplets, less and irregular microfilaments around the nucleus, broadened and increased extracelular matrix and the emergence of large gaps. These results demonstrate that under continuous mandibular advancement, the rat condylar cartilage wil become thick or thin with the endurance time, and chondrocyte matrix synthesis ability wil be significantly enhanced.

AIM: To evaluate cell proliferation, apoptosis and migration of human retinal pigment epithelial cells ( RPE) when co - cultured with human marrow mesenchymal stem cells ( hMSCs ) in condition of hypoxia and hyperglycemia so as to explore possible mechanisms of diabetes aggravating choroidal neovascularization ( CNV) preliminarily.
METHODS:Both hMSCs and RPE cells were co-cultured in a transwell system. The experiment was divided into four groups: 21% O2 with 5. 56mmol/L glucose ( control group, A ), 21% O2 with 30mmol/L glucose ( hyperglycemia and normoxia group, B ) , 5% O2 with 5.56mmol/L glucose ( normoglycemia and hypoxia group, C ) and 5% O2 with 30mmol/L glucose ( hyperglycemia and hypoxia group, D) . Cell Counting Kit-8 (CCK-8) was used to detect the proliferation of RPE cells in each group at 12, 24 and 48h respectively. Flow cytometry was performed to observe apoptosis of RPE cells at 24h. Additionally, we assessed migration
capabilities of RPE via transwell assay under the condition of hyperglycemia and hypoxia by co-culturing of hMSCs.RESULTS:In this co-culturing system, at 12, 24 and 48h, group B (1. 61±0. 41, 1. 80±0. 34;1. 91±0. 35), C (1.34±0. 46, 1. 94±0. 40, 2. 14±0. 41) and D (1. 98±0. 47, 2.26±0.42, 2. 55±0. 40) showed significantly higher proliferation rate than group A (0. 92±0. 45, 1. 27±0. 32, 1.59±0. 41, P<0. 05). The migration capabilities of RPE in group B (149. 5±9. 19), C (140±9. 90) and D (170. 5±7. 78) increased dramatically compared with group A ( 114. 5±7.78, P<0.05) at 24h, whereas there was no significant difference of apoptosis ratio among four groups (P>0. 05).
CONCLUSION:By coexistence with hMSCs, the synergy of hyperglycemia and hypoxia can improve migration and proliferation of RPE cells, and have no effect on apoptosis of RPE cells within short period.

Objective To assess the value of magnetic resonance diffusion weighted imaging(DWI) and apparent diffusion coefficients(ADC) values of hepatic alveolar echinococcosis(HAE) in children.Methods 20 cases of children(≤14 years) with HAE were collected in this restrospective study.PNM staging was determined, the HAE peripheral area of DWI lesions with different P stages was observed, and the ADC value of the peripheral area was measured.The comparison of alveococcus lesions in different stages of DWI with continuous edge degree and ADC value difference was done to evaluate the growth activity.Results There were 5 cases of P1 lesions, 7 cases of P2 lesions, 2 cases of P3 lesions and 6 cases of P4 lesions.DWI features of peripheral area were as follows: High signal ring band between HAE lesion edge and adjacent normal hepatic parenchyma was observed.P1 lesions showed almost complete obviously high signal peripheral area, indicating the most active proliferation, P2 and P3 lesions of peripheral area were continuous and with high signal, and still had obvious growth activity.P4 lesions of peripheral area were not continuous, while the signal decreased, indicating the activity also decreased.The highest ADC value was detected in P1 lesions group of and the ADC value of P2 lesions group were lower than P1, and the ADC value of P4 lesions group were the lowest.P3 lesions samples were too small and thus no statistical analysis was done.Differences of ADC value between the three groups were statistically significant (P<0.05).Conclusion DWI image features could be used to assesse the growth activity of HAE in children with different stages to a certain extent.ADC values measurement provides important reference value for evaluating the growth activity at various stages of the lesions.

In order to clarify the chemical constituents of Si-Wu Decoction rapidly and holistically, we analyzed the ethanol extract of Si-Wu Decoction by UPLC/Q-TOF-MSE and UNIFI which based on traditional Chinese medicine database, the probable structures of 113 compounds were identified. The results show that this method can rapidly and effectively characterize the chemical compounds of Si-Wu Decoction and provide a new solution for identification of components from complex TCM extract.

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.

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