BACKGROUND:Previous studies have found that miR-21 expression is increased during osteogenic differentiation of bone marrow mesenchymal stem cells, but the action and molecular mechanism of miR-21 are stil unclear. OBJECTIVE:To verify the target gene of miR-21, Spry1, and to explore the role of Spry1 in osteogenic differentiation of human bone marrow mesenchymal stem cells. METHODS:Luciferase report was used to verify Spry1 gene targeted by miR-21, and western blot assay was used to detect the expression of Spry1 in the osteogenesis of human bone marrow mesenchymal stem cells. Spry1 expression vector was established and transfected into human bone marrow mesenchymal stem cells. Osteogenesis ability of human bone marrow mesenchymal stem cells was analyzed after Spry1 high expression by alkaline phosphatase, alizarin red staining, RT-PCR and western blot. RESULTS AND CONCLUSION:Luciferase report suggested that Spry1 was a target gene of miR-21. The expression level of Spry1 was decreased in the osteogenesis of human bone marrow mesenchymal stem cells. Increasing expression of Spry1 could inhibit osteogenic differentiation of human bone marrow mesenchymal stem cells. These results indicate that Spry1 as a target gene of miR-21 negatively regulates osteogenic differentiation of human bone marrow mesenchymal stem cells, and plays an important role in bone formation process.

BACKGROUND:Osteogenic differentiation is a complex process involving transcriptional and post-transcriptional regulation by multiple signaling pathways, and the specific mechanisms remain unclear. It is of great significance to study the role of critical miRNAs in the osteogenic differentiation of bone marrow mesenchymal stem cells (BMSCs) in the treatment of osteoporosis and bone defects. OBJECTIVE: To explore the regulatory ability of miR-21/Sprouty1 function axis in the osteogenic differentiation of BMSCs in patients with postmenopausal osteoporosis. METHODS:BMSCs were isolated from healthy people (H-hBMSCs) and patients with postmenopausal osteoporosis (PMOP-hBMSCs), and their osteogenic ability was compared. Expression of miR-21 and Spry1 at gene and protein levels was detected by real-time RT-PCR and western blot assay, respectively. miR-21 expression was upregulated via transfection in PMOP-hBMSCs, and the osteogenic ability and Spry1 expression of the cells were detected, while real-time RT-PCR and western blot were used to detect the expression of osteogenic marker genes, Runx2 and Osterix. RESULTS AND CONCLUSION:Compared with H-hBMSCs, PMOP-hBMSCs osteogenic ability was weakened significantly, miR-21 expression decreased, and Spry1 expression increased, indicating an inhibition to the miR-21-Spry1 function axis. Through the transfection of miR-21 and down-regulation of Spry1, the expression levels of Runx2 and Osterix were increased, and PMOP-hBMSCs osteogenic ability was partially restored.

Adult ; Hamartoma ; pathology ; Humans ; Lung ; pathology ; Lung Neoplasms ; pathology

OBJECTIVETo study the pathology, diagnosis and differential diagnosis of papillary carcinoma of the breast featuring expansile invasion.

METHODSMorphologic analysis and immunohistochemical study using MaxVision method were carried out in 25 cases of papillary carcinoma of the breast featuring expansile type of invasion.

RESULTSAll the 25 cases were female, and ranged from 26 to 84 years old in age with median of 69. The diameters of the neoplasms were from 1 cm to 5 cm, among which those of two cases were 4 cm and 5 cm respectively and the others were less than 2.5 cm. The tumor appeared solid and well-demarcated on gross examination. Histologically, the tumor was non-encapsulated. In some cases, the tumor tissue was separated by fibrous septa or intervening native breast tissue. The tumor was composed of arborizing papillae with fibrovascular cores, associated glandular fusion and cribriform pattern. The fibrovascular cores were covered by cuboidal to columnar cells. In 3 of the cases studied, focal transition with micropapillary ductal carcinoma-in-situ was demonstrated. The tumor cells showed mild or moderate degree of nuclear pleomorphism and contained amphophilic to eosinophilic cytoplasm. Mitotic figures were not frequently seen. One case displayed squamous metaplasia and 4 cases showed apocrine metaplasia. Dimorphic features were identified in 2 cases. The stromal tissue within the tumor was often scanty and sometimes sclerotic, associated with various degrees of inflammatory infiltrate and hemosiderin deposition. Immunohistochemical study for smooth muscle actin, p63, CD10 and CK5/6 showed negative staining in all of the 25 cases studied, including the 2 cases with dimorphic features. Seven cases had breast cancer marker study performed. Six cases were positive for estrogen receptor and progesterone receptor. HER2 oncoprotein was not over-expressed in 6 cases. Nine patients underwent axillary dissection and 2 of them showed axillary nodal metastasis.

CONCLUSIONSDiagnosis of papillary carcinoma of the breast is one of the most difficult areas in breast pathology. Demonstration of expansile type of invasion, when coupled with complex papillary fusion, cribriform pattern and absence of myoepithelial cells on immunohistochemistry, is helpful in arriving at a correct diagnosis.

Adult ; Aged ; Aged, 80 and over ; Breast Neoplasms ; metabolism ; pathology ; surgery ; Carcinoma, Papillary ; metabolism ; pathology ; surgery ; Diagnosis, Differential ; Female ; Humans ; Lymph Node Excision ; Lymphatic Metastasis ; Mastectomy, Modified Radical ; Middle Aged ; Mucin-1 ; metabolism ; Neoplasm Invasiveness ; Receptors, Estrogen ; metabolism ; Receptors, Progesterone ; metabolism

OBJECTIVETo retrospectively study and analyse the immune regulatory effect of Bailing Capsule (BLC, a dry powder preparation of Cordyceps sinensis mycelia) on patients after renal transplantation, its influences on various systems of organism, and to explore its possible acting mechanism.

METHODSIn accordance with the entry criteria, 67 recipients of renal homo-allograft were assigned to two groups. The 42 cases in the control group were treated with mycophenolate mofetil (MMF) plus cyclosporine A (CsA), or tacrolimus (FK506) plus prednisone (Pred); the 25 in the treated group treated with the chemotherapy the same as in the control group plus BLC. They were followed up for 48 weeks by checking up blood routine, urine routine, hepatic and renal function, total serum protein, serum albumin, uric acid, etc., and the dosage of immunoinhibitory used was recorded periodically.

RESULTSComparison showed no significant difference in graft survival rate, occurrence of reject reaction and renal function recovery between the two groups; but levels of urinary erythrocytes and leucocytes, blood alanine transaminase, aspartate amino transferase, uric acid, total bilirubin, direct bilirubin, as well as the incidence of infection were significantly lower, and serum total protein and albumin were significantly higher in the treated group (all P < 0.01); moreover, counts of erythrocyte and leukocyte from 12 to 48 weeks, T-lymphocyte from 4 to 48 weeks after transplantation were significantly higher in the treated group (P < 0.05 and P < 0.01), and the recovery appeared earlier, the dosage of CsA or FK506 used 12 weeks after operation was significantly lower in the treated group than in the control group (P < 0.05, P < 0.01).

CONCLUSIONSBLC could effectively protect liver and kidney, stimulate hemopoietic function, improve hypoproteinemia, as well as reduce the incidence of infection and the dosage of CsA and FK506 used, etc. Therefore, it is a useful drug for immunoregulation after organ transplantation.

Adult ; Capsules ; Cordyceps ; Cyclosporine ; administration & dosage ; Drugs, Chinese Herbal ; therapeutic use ; Female ; Humans ; Kidney Transplantation ; Male ; Middle Aged ; Tacrolimus ; administration & dosage

OBJECTIVETo investigate the pathologic and immunohistochemical features, diagnosis and differential diagnosis of low-grade adenosquamous carcinoma of the breast and syringomatous adenoma of the nipple.

METHODSSix cases of low-grade adenosquamous carcinoma of the breast and four cases of syringomatous adenoma of the nipple were examined histologically and immunohistochemically (MaxVision method), and the literature was reviewed.

RESULTSThe two types of tumors were similar in morphology, but located in different regions with low-grade adenosquamous carcinoma being present in the deep parenchyma and syringomatous adenoma in nipple. Both types of tumors were composed mainly of well-differentiated glands with angulated, comma shaped or polliwog appearance in a disordered infiltrative pattern. The tumor cells also formed solid tubules, strips or nests, with frequent areas of squamoid differentiation. Mitosis was rare. The interstitial tissue showed abundant spindle cells or sclerotic fibrosis with mixed inflammatory cells infiltration. One case of low-grade adenosquamous carcinoma showed a concomitant malignant adenomyoepithelioma, and another case showed concomitant spindle cell metaplastic carcinoma. One case of syringomatous adenoma involved the deep parenchyma. Immunohistochemistry of low-grade adenosquamous carcinoma showed that CK5/6 and p63 were positive in the outer layer of the glands and the squamoid epithelium, and CD10 was also positive in the outer layer of the glands. ER and HER2 were negative, and PR was also negative except for one case in which the spindle cells were positive for CK5/6, AE1/AE3 and PR focally. Immunostaining of syringomatous adenoma demonstrated that p63 and CK5/6 were positive in the outer layer of the glands and the squamoid epithelium. Calponin, SMA, ER, PR and HER2 were all negative.

CONCLUSIONSLow-grade adenosquamous carcinoma of the breast and syringomatous adenoma of the nipple are similar in morphology and immunohistochemical phenotype, while the biological features are opposite due to different locations. The differential diagnoses include tubular carcinoma, adenosquamous carcinoma, radial sclerosing lesions and others.

Adenocarcinoma ; pathology ; Adult ; Aged ; Breast ; pathology ; Breast Neoplasms ; diagnosis ; metabolism ; pathology ; Carcinoma, Adenosquamous ; diagnosis ; metabolism ; pathology ; Carcinoma, Squamous Cell ; pathology ; Diagnosis, Differential ; Female ; Humans ; Immunohistochemistry ; Keratin-5 ; metabolism ; Keratin-6 ; metabolism ; Middle Aged ; Neprilysin ; metabolism ; Nipples ; pathology ; Sclerosis ; Sweat Gland Neoplasms ; diagnosis ; metabolism ; pathology ; Syringoma ; diagnosis ; metabolism ; pathology ; Transcription Factors ; metabolism ; Tumor Suppressor Proteins ; metabolism

Objective To explore the relationship between dynamic changes in ultra-micro-structural of pulmonary arteries and endogenous hydrogen sulfide in rats with left-right shunt.Methods Rats in shunt group were subjected to an abdominal aorta-inferior vena cava shunt to create an animal model of pulmonary artery structural remodeling. After 1 day, 3 days, 1 week, 4 weeks and 8 weeks of experiment, the ultra-micro-morphologic changes of pulmonary arteries of rats were observed under electronic microscope and H_2S concentration in serum was evaluated by modified sulfide electrode method.Results The changes of ultra-micro-structure of pulmonary arteries were progressively exacerbated, endothelial cells became swollen and large in size on 3 days, smooth muscular cells increased in size as well as the change of endothelial cells in 1 week, and they changed from contractile phenotype to synthetic phenotype in 4 weeks.Conclusions Shunt exhibited changes of ultra-micro-structure of pulmonary arteries are accompanied by the changes of endogenous H_2S. It is suggested that endogenous H_2S might play a protective role in changes of ultra-micro-structure of pulmonary artery.

Objective To evaluate the nutritional status of iodine of pregnant and lactating women in coastal and inland areas of Zhejiang province,and to provide scientific basis for iodine supplementation.Methods In 2009,five coastal cities,five coastal counties and three inland areas from Zhejiang province were selected by the cluster sampling method,and one sub-district(township) was randomly selected in each city (county),then one community (village) was randomly selected in each sub-district (township).Thirty pregnant women,thirty lactating women and fifty children aged 8 - 10,were randomly selected in each community(village) as the investigation objects to collect instant urine samples to detect urine iodine,and to collect thirty resident edible salt and some water samples to detect salt iodine and water iodine,if the numbers of water source were more than 5 then 1 water sample was collected by their location of east,south,west,north and center in each community(village),otherwise,the numbers of water samples collected were equal to the number of community (village).Results The medians of salt iodine of coastal city,coastal county and inland area were 28.42,25.29,and 33.24 mg/kg,respectively,and the medians of water iodine correspondingly were 5.41,11.52,and 2.80 μg/L,respectively.The medians of urinary iodine of pregnant and lactating women in Zhejiang province were 148.72 and 161.34 μg/L,respectively,which were less than that of children aged 8 - 10(231.02 μg/L,Z =8.57,8.24,all P ＜ 0.017).The medians of urinary iodine in pregnant women from different areas were ranked in order as inland areas(211.42 μg/L),coastal counties (150.51 μg/L) and coastal cities (123.33 μg/L,Z =2.80,5.31,2.42,all P ＜ 0.017),and in lactating women,the medians of urinary iodine in inland areas( 197.43 μg/L) was higher than that in coastal cities( 139.64 μg/L,Z =4.03,P ＜ 0.017).Conclusions The overall levels of iodine nutrition in lactating and pregnant women in Zhejiang province are in the appropriate level,but that of the pregnant women in the coastal cities is inadequate.

AIMTo develop an HPLC-ESI-MS assay for determination of trimebutine in human plasma and to investigate the pharmacokinetics and bioequivalence of two trimebutine tablets in human.

METHODSAfter being made alkaline with saturated sodium bicarbonate, plasma was extracted by cyclohexane and separated by HPLC on a reversed-phase C18 column with a mobile phase of 10 mmol x L(-1) ammonium acetate buffer solution (pH 3.5)-methanol (18:82). HPLC-ESI-MS was performed in the selected ion monitoring (SIM) mode using target ions at m/z 388 for trimebutine and m/z 280 for the internal standard (sibutramine, IS). The fragmentor voltage was 50 V. A randomized cross-over design was performed in 20 healthy volunteers. In the two study periods, a single 100 mg dose of each tablet was administered to each volunteer.

RESULTSCalibration curve was linear over the range of 0.3 - 150 microg x L(-1). The main pharmacokinetic parameters of T1/2, Tmax and Cmax were (9.2 +/- 2.8) h, (1.0 +/- 0.3) h and (40 +/- 20) microg x L(-1) for the reference tablet; (9.2 +/- 2.3) h, (0.9 +/- 0.4) h and (41 +/- 20) microg x L(-1) for the test tablet. The relative bioavalability of the test tablet was (97 +/- 13)%. The results of variance analysis and two one-sided t-test showed that there was no significant difference between the two formulations in the AUC and Cmax.

CONCLUSIONThe assay was proved to be sensitive, accurate and convenient. The two formulations were bioequivalent.

Adult ; Area Under Curve ; Chromatography, High Pressure Liquid ; Gastrointestinal Agents ; administration & dosage ; pharmacokinetics ; Humans ; Male ; Spectrometry, Mass, Electrospray Ionization ; Tablets ; Therapeutic Equivalency ; Trimebutine ; administration & dosage ; pharmacokinetics

OBJECTIVETo investigate the effect of chemically synthetic small interfering RNA (siRNA) targeting connective tissue growth factor (CTGF) on the synthesis and secretion of extracellular matrix (ECM) in hepatic stellate cells (HSC).

METHODSChemically synthetic siRNA targeting CTGF was transfected into HSC T6 (an active HSC line in rats) by oligofectamine package, and untreated HSC T6 were used as control. Total RNA and protein of the cells, after their incubation with siRNA for 24, 48 and 72 hours, were extracted, and the supernatants were collected. The expressions of CTGF and type I and III collagen genes were detected by means of reverse transcription-polymerase chain reaction (RT-PCR) and/or Western blot. Contents of hyaluronic acid and type III pro-collagen in the supernatants were determined by radioimmunoassay.

RESULTSThe expression of CTGF at mRNA and protein level and type I and III collagen at mRNA levels were markedly down-regulated in siRNA-transfected HSCs. The contents of hyaluronic acid and type III pro-collagen in the supernatants decreased by 46%+/-7%, 52%+/-7%, 53%+/-7% and 29%+/-18%, 29%+/-7%, 27%+/-5%, compared with those of the blank control at 24, 48 and 72 hours.

CONCLUSIONSChemically synthetic anti-CTGF siRNA can significantly inhibit CTGF gene expression in HSC, and markedly reduce the synthesis and secretion of ECM including type I and III collagen and hyaluronic acid. The siRNA-directed suppression of CTGF gene in HSC was maintained for 72 hours. This suggests that chemically synthetic siRNA may be a potential in preventing and treating liver fibrosis and may have a promising future for development

Cell Line ; Connective Tissue Growth Factor ; Extracellular Matrix ; metabolism ; Gene Targeting ; Humans ; Immediate-Early Proteins ; genetics ; Intercellular Signaling Peptides and Proteins ; genetics ; Liver ; cytology ; metabolism ; RNA, Small Interfering ; genetics