AIM:To research the effects of lithium chloride on transforming growth factor beta (TGF-β) and connective tissue growth factor (CTGF) in cultured human Tenon capsule fibroblasts (HTFs) and explore its mechanism.METHODS:HTFs were cultured and identified by vimentin staining with immunofluorescence and the morphological characteristics.The experimental group was processed 48h with LiCl in concentration of 80mmol/L, the control group without LiCl.The mRNA expression of TGF-β and CTGF in two groups were analyzed with real-time fluorescent quantitative polymerase chain reaction (real time-qPCR) and the protein expression was detected with Western blot.RESULTS:The cultured HTFs expressed TGF-β and CTGF.The mRNA expression of TGF-β and CTGF significantly decreased compared with the control group(t=20.042, 14.995, P<0.05).the protein expression of TGF-β and CTGF also decreased significantly compared with the control group(t=46.058、12.452, P<0.05)CONCLUSION:The cultured HTFs can express TGF-β and CTGF in mRNA and proteins' level.LiCl can reduce the expression of TGF-β and CTGF both in gene and proteins' level.LiCl has the potential to modulate wound healing for glaucoma filtration surgery.

Objective To investigate the CT and MRI findings of primary hepatic neuroendocrine carcinomas.Methods Imaging findings of 6 patients with pathologically confirmed hepatic neuroendocrine carcinomas were retrospectively analyzed.Four patients underwent plain CT and contrast enhanced CT examinations.The other 2 patients underwent plain MRI and contrast enhanced MRI.Results One out of the 6 cases manifested multicentric tumor which appeared as one large tumor surrounded with multiple small nodules peripherally,and the tumors of the other 5 cases were solitary.On plain CT images,all lesions were well-defined and hypoattenuating with central areas of even lower density except punctuate calcifications at tumor periphery in 1 case.On pre-contrast MRI,the cases manifested heterogeneous low signal intensity on T1 wI and slightly high intensity on T2 WI.On dynamic contrast enhanced CT and MRI,Tumors manifested slight to mild enhancement in arterial phase,slight enhancement in portal venous phase or delayed phase with no enhancement in the center,accompanied by dislocation of surrounding vessels.There Was no lymphadenopathy in peritoneal cavity and retroperitoniurm. Conclusion CT and MRI scanning can demonstrate specific features of primary hepatic neuroendocrine carcinoma,which may help the diagnosis and differential diagnosis of the tumor.

AIMTo introduce the manufacture and use of a simple, new type of drop recorder of frog heart.

METHODSTo improve the perfusion device of (see text for symbol) and Straub method. Two electrodes of drop recorder were fixed in an injector of 20 ml. The input tube, output tube and resistance tube were all made of plastic material.

RESULTSThis device could be used to observe effects of preload, after-load, hormone and electrolyte on the cardiac output in isolated frog heart.

CONCLUSIONThe new type of drop recorder was economical and could be easily operated, it could be also connected to computer. Using the new type of drop recorder, effects of various physical and chemical factors on cardiac function could be observed directly, accurately.

Animals ; Electrodes ; Heart ; physiology ; Infusion Pumps ; Ranidae ; Weights and Measures ; instrumentation

OBJECTIVETo observe blood uric acid levels and Goldstein grading, as well as their correlation in Wilson's disease (WD) patients with different Chinese medical syndrome types.

METHODSTotally 906 WD patients in line with inclusive criteria were assigned to 6 groups, i.e., the heart spirit confused by phlegm group (HSCP, 26 cases), the phlegm-fire disturbing heart group (PFDH, 90 cases), the retention of damp-heat group (RDH, 113 cases), deficiency of qi and blood group (DQB, 168 cases), the deficiency of Gan-yin and Shen-yin group (DGYSY, 327 cases), the deficiency of Gan and Shen group (DGS, 182 cases) due to different Chinese medical syndrome types. Recruited were another 160 healthy subjects having similar ages and diet structures, who came for medical examinations, as the healthy control group. Venous blood was collected from the medial cubital vein of each-patient on an empty stomach in early mornings to detect blood uric acid levels. Results Blood uric acid levels were lower in each syndrome type group than in the healthy control group (146.08 +/- 67.24 micromol/L in the HSCP group; 157.08 +/- 69.77 micromol/L in the PFDH group; 162.58 +/- 97.72 micromol/L in the RDH group; 156.20 +/- 62.63 micromol/L in the DQB group; 161.83 +/- 111.23 micromol/L in the DGYSY group; 194.41 +/- 90.01 micromol/L in the DGS group; 242.39 +/- 87.55 micromol/L in the healthy control group, P < 0.01). Blood uric acid levels were higher in the DGYSY group than in the other 5 syndrome groups (P < 0.01). Correlation analyses between Goldstein grading and blood uric acid showed that, along with increased Goldstein grade (that was aggravating disease conditions), WD patients' blood uric acid levels decreased (P < 0.01).

CONCLUSIONSWD patient's blood uric acid levels decreased more. Blood uric acid levels and Goldstein grading were different in various Chinese medical syndrome types. Blood uric acid levels had certain value in assessing the severity of WD.

Asian Continental Ancestry Group ; Heart ; Hepatolenticular Degeneration ; blood ; classification ; diagnosis ; Humans ; Medicine, Chinese Traditional ; Syndrome ; Uric Acid ; blood

OBJECTIVESTo explore the changes of hepatic gene expression during the course of nonalcoholic fatty liver disease (NAFLD) development in rats.

METHODSA rat model of NAFLD was developed by feeding the animals a high-fat diet for 24 weeks. Liver tissues of the model rats and the control rats were analyzed at different time points using rat U230A (Affymetrix GeneChip), which covers 15650 genes.

RESULTSCompared with the control rats, the number of genes expressed differently in the model group rats at 4 and 8 weeks was 426 and 540. The up-regulated genes among them were intracellular phosphorylase genes, metabolic enzyme genes, fatty acid binding protein genes, cytochrome P450 genes, cellular transcription and differentiation genes. The down-regulated genes were ionic channel genes, hormone receptor genes, and cytoskeleton genes. At the 12th week, the number of the genes expressed differently was 501, in which 352 were up-regulated genes, including genes related to inflammation and apoptosis such as interleukin and Toll-like receptor 4. At the 16th week, the number of the differently expressed genes was 665, with 430 up-regulated, such as those related to the inflammation and apoptosis genes and collagen I and fibrosis genes, however cell regeneration genes were down-regulated. At the 24th week the number was 663, of which fibroblast growth factor, transforming growth factor and insulin-like growth factor genes were up-regulated. Of all the differently expressed genes, the number of up-regulated genes was 128, including 10 lipogenic genes, 46 metabolic genes, 15 inflammation genes, 10 apoptosis genes, and 16 fibrosis genes; and the down-regulated genes were 52, including 6 hormone receptor genes, 5 cell regeneration genes and 11 electron transport genes.

CONCLUSIONThe changes of the hepatic gene expression of rats fed a fat-rich diet are related to the duration of the feeding, and are correlated with their histopathology in the livers.

Animals ; Dietary Fats ; Fatty Liver ; etiology ; genetics ; Gene Expression Profiling ; Liver ; metabolism ; Oligonucleotide Array Sequence Analysis ; Rats ; Rats, Sprague-Dawley

Fatigue is a common complication of type 2 diabetes mellitus (T2DM). We examined the relationship between T2DM fatigue and the skeletal muscle 5-hydroxytryptamine (5-HT) system. In animal experiments, a T2DM model was established in mice by feeding a high-fat diet with intraperitoneal injection of streptozotocin. The mice were treated with the 5-HT_2A receptor antagonist sarpogrelate hydrochloride (SH) and the 5-HT synthesis inhibitor carbidopa (CDP) (separately and in combination). In cell culture experiments, C2C12 cells were stimulated with D-glucose, palmitic acid or 5-HT. 5-HT_2AR, 5-HT synthesis and 5-HT degradation were inhibited by SH, CDP, or monoamine oxidase A (MAO-A) inhibitor. The animal experiments were in accordance with the regulations of the Animal Ethics Committee of China Pharmaceutical University. The results showed that 5-HT_2AR, 5-HT synthase and MAO-A were expressed in mouse skeletal muscle and C2C12 cells. The expression of these proteins was significantly up-regulated in T2DM mice or when C2C12 cells were exposed to palmitic acid and D-glucose; palmitic acid was a stronger stimulant of their expression than D-glucose. Rotating rod experiments and biochemical index tests have shown that T2DM fatigue is associated with an increase in skeletal muscle 5-HT_2AR, 5-HT synthesis and 5-HT degradation. 5-HT_2AR mediates the expression of MAO-A and the synthesis of 5-HT, which indirectly regulates the degradation of 5-HT. MAO-A regulates cell inflammation, mitochondrial ROS production and membrane potential depolarization by mediating 5-HT degradation. MAO-A also inhibits the expression of peroxisome proliferator-activated receptor γ coactivator-1 (PGC-1), carnitine palmitoyltransferase-1 (CPT1) and ATP synthase-6 (ATP6), thus inhibiting mitochondrial functions such as fatty acid β oxidation and ATP synthesis. SH and CDP can effectively treat T2DM fatigue, and can also reduce blood glucose and blood lipid, and the combination of SH and CDP has a clear synergistic effect.

OBJECTIVETo investigate the number of peripheral blood CD5(+) B cells and their ability of secreting IL-10 in patients with immune thrombocytopenia (ITP).

METHODSPeripheral blood lymphocytes were isolated from 57 pre-treated, 40 post-treated ITP patients and 25 controls using Ficoll-Hypaque density centrifugation and then stained with PE-CD5/FITC-CD19 for flow cytometric analysis. After 24-hour culture, lymphocytes were stained with APC-IL-10 for intracellular cytokine detection. ELISA assay was employed to determine IL-10 concentration in supernatants.

RESULTSThe percentage and absolute number of CD5(+) B cells in peripheral blood from pre-treated ITP patients were significantly higher than that from normal controls (3.75 ± 2.37)% vs (2.10 ± 1.08)%, P < 0.01; (6.29 ± 5.77)× 10(7)/L vs (3.06 ± 1.90)× 10(7)/L, P < 0.01. CD5(+) B cells expressed more intracellular IL-10 than other lymphocyte subsets both in ITP patients and normal controls. The percentages of IL-10(+) cells within CD5(+) B cells in pre-treated ITP patients and normal controls were (29.51 ± 20.73)% and(15.90 ± 9.58)%, respectively(P < 0.01). Intracellular mean fluorescence intensity (MFI) of IL-10 in CD5(+) B cells was 27.95 ± 13.99 in pre-treated patients, which was significantly higher than that in controls (P < 0.01). In contrast, IL-10 concentration in supernatants was (173.05 ± 102.50) ng/L in pre-treated ITP group, which was lower than that (230.61 ± 76.96) ng/L in controls. In patients who achieved remission, the number of CD5(+) B cells decreased to level comparable to normal controls. While intracellular IL-10 MFI of CD5(+) B cells in post-treated ITP patients remained as high as in pre-treated ones, the IL-10 concentration in supernatants increased to level similar to controls.

CONCLUSIONThe significantly increased number of CD5(+) B cells and accumulated IL-10 in CD5(+) B cells suggested impaired IL-10 secretion in ITP patients. The number and the ability of secreting IL-10 of CD5(+) B cells could be restored after effective treatments in patients with ITP.

Adult ; Aged ; B-Lymphocytes ; immunology ; metabolism ; CD5 Antigens ; metabolism ; Case-Control Studies ; Female ; Humans ; Interleukin-10 ; blood ; Male ; Middle Aged ; Purpura, Thrombocytopenic, Idiopathic ; blood ; immunology ; Young Adult

A GeXP based multiplex PCR assay was developed to simultaneously detect six different chicken respiratory viruses including H5, H7, H9 subtypes of avian influenza virus(AIV), new castle disease virus (NDV), infectious bronchitis virus(IBV) and infectious laryngotracheitis virus(ILTV). According to the conserved sequences of genes of each pathogen, seven pairs of specific primers were designed, and the reaction conditions were optimized. The specificity and accuracy of GeXP were examined using samples of single and mixed infections of virus. The sensitivity was evaluated by performing the assay on serial 10-fold dilutions of cloned plasmids. To further evaluate the reliability, thirty-four clinical samples were detected by GeXP. The corresponding specific fragments of genes were amplified. The detection limit of GeXP was 10(2) copies/microL when all of 7 pre-mixed plasmids containing target genes of six chicken respiratory viruses were present. In the detection of thirty-four clinical samples, the results of GeXP were accorded with the viral isolation completely. In conclusion, this GeXP assay is a rapid, specific, sensitive and high-throughput method for the detection of chicken respiratory virus infections. It can be applied in rapid differential diagnosis for clinical samples, and also provide an effective tool to prevent and control chicken respiratory diseases with similar clinical symptoms.
Animals ; Chickens ; Influenza A virus ; classification ; genetics ; isolation & purification ; physiology ; Influenza in Birds ; diagnosis ; virology ; Multiplex Polymerase Chain Reaction ; methods ; Poultry Diseases ; diagnosis ; virology ; Respiratory Tract Infections ; diagnosis ; veterinary ; virology

Objective To explore the influence of cyasterone on the osteoclast and osteoblast differentiation and then to investigate its effect on the bone quality in the osteoporosis mice. Methods CCK8 assay was firstly used to detect the toxic effect of cyasterone on the mouse bone marrow derived mononuclear macrophages (BMMs) and anterior osteoblast lines MC3T3E1. Cell apoptosis was measured by flow cytometry. Then TRAP staining and ALP staining were employed to detect osteoclast differentiation and osteoblast differentiation, respectively. Realtime PCR was carried out to test the expression of osteoclast special gene TRAP and osteogenesis crucial gene ALP. In vivo, 15 mice were divided into three groups: sham-operated group, OVX group and OVX+cyasterone treatment group. In treatment group, cyasterone was used as 5mg/kg every day. Sham-operated group and OVX group were treat with saline solution. After 4 weeks, the tibia was collected for Micro-CT detection to observe the bone quality and microstructure changes. Results Cyasterone with the concentration of less than 10 mg/L had no significant cytotoxicity nor influence on the apoptosis (P＞0.05) . Cyasterone could significantly inhibit the osteoclast differentiation of BMMs (P＜0.05), simultaneously, it also had the effect to promote the osteoblast differetiation of MC3T3E1. Real-time PCR indicated that cyasterone could block the expression of TRAP and increase the expression of ALP (P＜0.05) . In vivo, cyasterone was able to obviously improve the osteoporosis status caused by estrogen deficiency without general toxicity. Conclusion cyasterone could provide a good treatment for osteoporosis through the bidirectional effect of inhibiting osteoclast differetiation and promoting osteoblast differentiation.

OBJECTIVETo assess the association between 2 single nucleotide polymorphisms (SNPs) of ETS1 gene and susceptibility to systemic lupus erythematosus (SLE) in a northern Chinese Han population.

METHODSTwo SNPs within the ETS1 gene mapped to 11q23 were selected based on HapMap data. Genotyping was conducted with Taqman method in 231 patients with SLE and 474 healthy controls from Qilu Hospital, Shandong and analyzed with PLINK1.07 software. Haplotypes were analyzed with SHEsis software.

RESULTSA statistically significant difference was detected in the distribution of rs1128334 and rs4937333 genotypes between the two groups (all P< 0.01). For rs1128334, the frequency of the minor allele was 0.291 and 0.428 in controls and cases, respectively. For rs4937333, the minor allele frequency was 0.381 and 0.476 in controls and cases respectively. An A-C haplotype was found to be strongly associated with increased risk for SLE, while another haplotype G-C may reduce this risk.

CONCLUSIONOur study has suggested that rs1128334 and rs4937333 are strongly associated with the risk for SLE in northern Chinese Han population.

3' Untranslated Regions ; Adolescent ; Adult ; Aged ; Asian Continental Ancestry Group ; ethnology ; genetics ; Female ; Genetic Association Studies ; Humans ; Lupus Erythematosus, Systemic ; ethnology ; genetics ; Male ; Middle Aged ; Polymorphism, Single Nucleotide ; Proto-Oncogene Protein c-ets-1 ; genetics ; Young Adult