OBJECTIVETo observe the improvement of thyroid function and changes of Akt, p-Akt, mammalian target of rapamycin (mTOR), and para-mTOR (p-mTOR) expression in Graves' disease (GD) mice after intervened by Jiakangning Capsule (JC), and to explore possible mechanism for JC in treating GD.

METHODSGD model was established by immunizing female BALB/c mice with thyroid stimulating hormone receptor A subunit (Ad-TSHRα-289). Totally 70 successfully modeled mice were divided into the model group (n =20), the JC intervened group (n =25), the Methimazole Tablet intervened group (n =25) according to random digit table. A normal control group (n =15) and a vehicle control group (n =20, injected with Ad-null) were also set up. Mice in the JC intervened group were administered with JC suspension at the daily dose of 1. 5 g/kg by gastrogavag. Mice in the Methimazole intervened group were administered with Methimazole suspension at the daily dose of 2. 5 g/kg by gastrogavage. Equal volume of normal saline was administered to mice in the rest 3 groups by gastrogavage. All intervention lasted for 5 weeks. Six mice were selected from each group to observe pathological changes of thyroid tissues. Serum levels of thyroxine (T4), triiodothyronine (T3), thyroid stimulating hormone (TSH), and thyrotropin receptor antibody (TRAb) were analyzed by radioimmunoassay. Expression levels of Akt, p-Akt, mTOR, and p-mTOR in thyroid tissues were etermined by Western blot.

RESULTS(1) The thyroid gland in the GD model group showed proliferative changes, with enlarged follicles of various sizes. Interstitial stroma was filled with blood vessels. Structures of thyroid tissues in the JC intervened group and the Methimazole intervened group were significantly restored, and follicular hyperplasia was relieved. (2) Compared with the normal control group and the vehicle control group, levels of TRAb, T4, and T3 increased; ratios of P-Akt/β-actin, p-Akt/Akt, p-mTOR/β-actin, and p-mTOR/mTOR also increased in the model group (all P <0. 01). Compared with the model group, levels of TRAb, T4, and T3 decreased in the JC intervened group and the Methimazole intervened group (P <0. 01); ratios of p-mTOR/β-actin and pmTOR/mTOR decreased in the JC intervened group (P <0.01); ratios of P-Akt/β-actin, p-Akt/Akt, p-mTOR/β-actin, and p-mTOR/mTOR decreased in the Methimazole intervened group (P <0. 05, P <0. 01). Conclusion JC could reduce thyroid hormonc levels of GD mice and lower expression levels of mTOR, and its mechanism for improving thyroid function of GD mice might be associated with this influence.

Actins ; Animals ; Capsules ; Disease Models, Animal ; Drugs, Chinese Herbal ; pharmacology ; therapeutic use ; Female ; Graves Disease ; drug therapy ; pathology ; Methimazole ; Mice ; Mice, Inbred BALB C ; Receptors, Thyrotropin ; Signal Transduction ; TOR Serine-Threonine Kinases ; Thyrotropin ; Thyroxine ; Triiodothyronine

A in vitro platelet aggregation bioassay was developed for the quality control of XST capsules. The in vitro anti-platelet aggregation effect in rats was observed to detect the bioactivity of XST capsules. Panax notoginseng saponins and Xuesaitong lyophilizedpowder for injection were taken as standard control substances to determine the potency. According to the results, XST capsules showeda significant inhibitory effect on thrombin-induced platelet aggregation in a dose-dependent manner. The in vitro anti-platelet activity oflyophilized powder for injection was stabler than that of Panax notoginseng saponins, and so suitable to serve as a standard control substance. The biological potency of XST capsules compared with standard control substance was detected by using parallel line assay. According to the results, the established bioassay method had a good repeatability (RSD 2.92%). The sample test results could pass thereliability test(linear deviation P > 0.05, parallel deviation P > 0.05). This bioassay method could be used as one of the complementary quality control methods for XST capsules.
Animals ; Capsules ; pharmacology ; Drugs, Chinese Herbal ; pharmacology ; Male ; Panax notoginseng ; chemistry ; Platelet Aggregation ; drug effects ; Platelet Aggregation Inhibitors ; pharmacology ; Rats ; Rats, Sprague-Dawley ; Saponins ; pharmacology

Objective To study the plasma folate concentrations in the third trimester of pregnant women and newborn babies so as to assess the association between them.Methods Pregnant women in Yuanshi and Laoting counties in Hebei province from May to June in 2009 were recruited with related information collected at enrollment.Those pregnant women being enrolled were followed up until delivery.Maternal blood was collected before delivery,and cord blood was collected after the expulsion of the placenta.Data from 437 pairs of women and newborns were analyzed.Plasma folate concentration was measured by Microbiological assay,with maternal plasma folate concentration ＜6.8 nmol/L defined as folate deficiency.Neonatal plasma folate concentration below 10％ was defined as relative deficiency.Student t-test and ANOVA were used to compare the plasma folate concentrations between the groups and x2 test was used to compare the situation of folate deficiency.In order to assess the association between maternal and newborn folate levels,logistic regression analysis was used to estimate the odds ratio of the neonatal plasma folate relative deficiency between the maternal folate deficient and normal groups after adjusting factors as age,BMI,region,career and education.Linear regression was used to test the trend by quintiles of maternal plasma folate concentration.Pearson' s test was used to test the relationship between the ratio of neonatal and maternal plasma folate level and the level of maternal plasma folate.Results The geometric mean of maternal plasma folate concentration was 8.0(95％CI:7.6-8.5) nmol/L and the deficiency was 29.3％,but in newborn babies,they were 24.0(95％CI:23.1-25.0) nmol/L and 0.9％ respectively.The plasma folate level in newborn babies was 3.0 times as high as in maternal (t=32.519,P＜0.01 )but the neonatal plasma folate deficiency status was higher than in matemal ( x2=137.2,P＜0.01 ).When compared with the normal plasma folate level group,the risk on neonatal plasma folate relative deficiency in the maternal folate deficiency group was significantly higher aiter adjusted for confounders (OR=1.96,95％CI:1.02-3.80).The neonatal plasma folate level significantly increased along with the maternal plasma folate level (Ptrend＜0.05).The ratio of neonatal and maternal plasma folate level was significantly inversely correlated with the maternal folate level (r=-0.810,P＜0.001 ).Conclusion Folate status in newborns was much better than in their mothers',in the northern rural areas of China.The maternal folate status was positively correlated with their offspring' s.Active placental transport for folate was significantly increasing when the maternal plasma folate level decreased.

Objective To evaluate the pattern of energy metabolism and nutrients intake in patients with chronic viral hepatitis and posthepatitic cirrhosis to effectively direct their nutrition therapy.Methods Resting energy expenditure (REE) was measured with open-circuit indirect Jorimetry in 60 patients with chronic viral hepatitis and 60 patients with posthepatitic cirrhosis.Their normal basal energy expenditure (BEE) was predicted by Harris-Benedict equation and energy intake (EI) was determined by diet recall. Correlation between REE and indicators for nutrition assessment was analyzed.Results REE was (77? 21) kJ?kg~(-1)?d~(-1) in 60 patients with pusthepatitic cirrhosis,significantly lower than BEE[(95?16) kJ? kg~(-1)?d~(-1)(P0.05,and their EI was (127?34) kJ?kg~(-1)?d~(-1),1.41?0.43 times as REE,in which PROI was (1.02?0.29) g?kg~(-1)?d~(-1),1.31?0.61 times as PROE (0.87?0.34) g?kg~(-1)?d~(-1),also indicating a negative nitrogen balance (-2.02?4.07).REE,EI and intake of three nutrients,serum level of albumin and prealbumin (PA) and body weight significantly decreased in patients with posthepatitic cirrhosis,as compared to those in patients with chronic viral hepatitis (P

Objective To investigate the expression of the interleukin-1 receptor(IL-1R)Ⅰ,IL-1RⅡand IL-1R accessory protein(IL-1RAcP)in osteoarthritis and analyse their biological significance.Methods Immunohistochemistry and reverse transcription-polymerase chain raction(RT-PCR)were adopted to detect the expression of IL-1RⅠ,IL-1RⅡand IL-1RAcP on the synovium of 107 OA patients.Results Immunohis- tochemistry showed strong positive expression of IL-1RⅠand IL-1RAcP,and positive expression of IL-1RⅡ. The expression was distributed in lining cells,monocyts and vascular endothelial cells of the sublining area, but all of them were negative or weak positive in normal synoviums.RT-PCR showed the expression of IL-1RⅠ,IL-1RⅡand IL-1RAcP in OA synoviums was significantly enhanced than normal synoviums (P＜0.05),and the expression of IL-1RⅠwas significantly enhanced than IL-1RⅡ(P＜0.05),but no sig- nificant difference with IL-1RAcP(P＞0.05).In stageⅡandⅢOA synoviums,the expression of IL-1RⅠand IL-1 RAcP had no significant difference with normal synoviums(P＞0.05).The expression of IL-1RⅡin stageⅢOA synoviums was significantly enhanced than normal(P＜0.05).Conclusion IL-1RⅠ,IL-1RⅡand IL-1RAcP play significant roles in the pathogenesis of OA,especially IL-1RⅠand IL-1RAcP.But their increase is only observed in the early stage of OA.These suggest that they may have no association with the development of OA and have no direct association with the severity of OA.OA can be cured by interrupting the signal transduction path in which IL-1 has played biological roles.

Purpose To investigate the difference of expression of autophagy-related gene (Beclin1,LC3,mTOR) in the development of esophageal squamous cell cancer.Methods Immunohistochemical EnVision method was adopted to detect the expression of autophagy-related gene Beclinl,LC3 and mTOR in 30 cases of normal esophageal mucosa,32 cases of low-grade intraepithelial neoplasia (LGIN),34 cases of highgrade intraepithelial neoplasia (HGIN),35 cases of early carcinoma and 126 cases of advanced esophageal carcinoma,respectively.The correlation between their expression with clinicopathologic factors was also analysed.Results The expression of Beclin1 in advanced esophageal carcinoma was obviously higher than that in another four groups (P ＜ 0.005).LC3 expression in advanced esophageal carcinoma was significantly higher than that in normal esophageal mucosa,LGIN and early carcinoma (P ＜ 0.005).The expression of mTOR in advanced esophageal carcinoma was significantly higher than that in normal esophageal mucosa,LGIN and HGIN (P ＜ 0.005).In advanced esophageal carcinoma group,the expression of Beclin1,LC3 and mTOR was related to tumor TNM stage and lymph node metastasis (P ＜ 0.05).Beclin1 expression was positively associated with LC3 and mTOR expression in advanced squamous cell carcinoma (P ＜ 0.05).Positive correlation was also observed between the expression of mTOR and LC3 in advanced esophageal carcinoma and HGIN (P ＜ 0.05).Conclusion In the carcinogenesis and development of esophageal cancer,Beclin1,as a tumor suppressor gene,activates autophagy and leads to excessive self consumption and death of tumor cells.mTOR promotes tumor growth by inhibiting autophagy and promoting angiogenesis.The combined detection of Beclinl,LC3 and mTOR may be beneficial to evaluate the progression and prognosis of esophageal squamous cell cancer.

OBJECTIVETo investigate the genetic polymorphism of human cytomegalovirus (HCMV) glycoprotein H (gH) from infantile clinical isolates, to analyze the genotypic distribution of gH in different diseases of HCMV infection and try to find the correlations between the diseases and genotypes.

METHODFresh urine specimens were collected from the hospitalized children with different diseases whose blood HCMV-IgM and HCMV-IgG were positive. Virus was isolated from these specimens. Glycoprotein H of harvest clinical isolates was genotyped by nested-PCR combined with restriction fragment length polymorphism (RFLP), the purified PCR products were digested by restriction endonuclease HhaI. The digested products were genotyped by polyacrylamide gel electrophoresis and silver staining. Classification and results of sequencing were compared.

RESULTTotally 102 HCMV clinical isolates were obtained. Glycoprotein H gene of these clinical isolates (43 cases had infantile hepatitis syndrome, 38 cases had anicteric hepatitis, 13 pneumonia, 7 thrombocytopenic purpura, and 1 congenital CMV infection) were positive by nested-PCR, whose positive rate was 100%. The results showed that 62 strains were gH1 genotypes (60.8%), while 40 strains were gH2 (39.2%), mixed type or new genotype was not observed. In infantile hepatitis syndrome (26 clinical isolates were gH1 genotypes, 17 clinical isolates were gH2 genotypes), anicteric hepatitis (25 were gH1, 13 were gH2) and pneumonia (9 were gH1, 4 were gH2), the distribution of HCMV gH genotypes of infantile clinical isolates was consistent with the overall trend (χ(2) = 0.357, P > 0.05). However , the gH2 was more common than gH1 in the clinical isolates of patients with thrombocytopenic purpura (6 were gH2, 1 were gH2, χ(2) = 6.083, P < 0.05).

CONCLUSIONGenotype 1 was the dominant genotype of glycoprotein H in HCMV clinical isolates from our hospital infants. There was no significant difference between the distribution of gH genotypes in infantile hepatitis syndrome, anicteric hepatitis and pneumonia. However, gH2 was the dominant genotype in thrombocytopenic purpura. These findings suggested that there may be a certain relevance between gH genotype and different clinical manifestations.

Amino Acid Sequence ; Base Sequence ; Child, Preschool ; Cytomegalovirus ; classification ; genetics ; isolation & purification ; Cytomegalovirus Infections ; virology ; DNA Primers ; DNA, Viral ; genetics ; Female ; Genotype ; Hepatitis ; virology ; Humans ; Infant ; Infant, Newborn ; Male ; Pneumonia, Viral ; virology ; Polymerase Chain Reaction ; Polymorphism, Genetic ; Polymorphism, Restriction Fragment Length ; Urine ; virology ; Viral Envelope Proteins ; genetics

OBJECTIVETo study the effects of sodium tanshinone II A sulfonate (STS) on proliferation of fibroblasts (Fbs) in human hypertrophic scar (HS), the mRNA and protein expressions of transforming growth factor beta 1 (TGF-β1) and alpha smooth muscle actin (α-SMA), and to investigate the scar inhibition mechanism of STS.

METHODSFbs were isolated from HS tissues that were removed from eight patients after burn injury, and they were cultured in vitro. Cells from the 3rd to the 6th passages were used in the experiment. Fbs were divided into control group and experimental group according to the random number table, and cells in the experimental group was divided into 0.050, 0.075, 0.100, 0.125, 0.150, 0.200 mg/mL STS subgroups. Cells in each subgroup were cultured with the corresponding concentration of STS, and cells in control group were cultured in equal volume of serum-free medium. After being cultured for 24 and 48 h, cell morphology was observed with inverted phase contrast microscope; cell proliferation was determined with MTT method and the proliferation inhibition rate (IR) was calculated. After being cultured for 48 h, the protein levels of TGF-β1 and α-SMA were determined with Western blotting; the mRNA expressions of TGF-β1 and α-SMA were determined with RT-PCR (no 0.200 mg/mL STS subgroup was set for these two indicators). Data were processed with factorial analysis of variance; differences between groups were processed with LSD test or Games-Howell test for unequal variances.

RESULTS(1) Fbs grew well in control group, but reduction in adherence and disorderly arranged Fbs were observed in experimental group. The cells in experimental group became smaller and round, with increasing intracellular particles and necrosis. A large amount of necrotic debris of cells was observed in 0.200 mg/mL STS subgroup. (2) The absorbance value of Fbs in each experimental subgroup was significantly lower than that in control group (with P values all below 0.01). Along with the increase in the concentration of STS and extension of culture time, the IR value increased, showing a certain degree of time-concentration dependence. After being cultured with STS for 24 and 48 h, IR values of cells in the experimental subgroups were respectively 23.58%, 32.11%, 37.56%, 57.98%, 79.53%, 96.69% and 34.72%, 38.48%, 47.62%, 64.40%, 89.70%, 98.01%. (3) Except for the 0.050 mg/mL STS subgroup, the protein levels of TGF-β1 and α-SMA in the other subgroups were significantly lower than those in control group (with F values respectively 57.674, 47.795, P values all below 0.001). The protein levels of TGF-β1 and α-SMA reached the nadir in 0.150 mg/mL STS subgroup, respectively 0.34 ± 0.06, 0.33 ± 0.07. The relative expression amounts of TGF-β1 and α-SMA mRNA in the experimental subgroups were obviously decreased compared with those in control group (with F values respectively 68.548, 47.522, P values all below 0.001), which was most significant in 0.150 mg/mL STS subgroup, with TGF-β1 mRNA and α-SMA mRNA respectively 0.39 ± 0.07 and 0.42 ± 0.08.

CONCLUSIONSSTS can inhibit the proliferation of Fbs, reduce the protein and mRNA expressions of TGF-β1 and α-SMA, which may be beneficial to ameliorate the formation and contracture of HS, and it is assumed as a potential drug for treating scars.

Actins ; genetics ; metabolism ; Adult ; Cell Proliferation ; drug effects ; Cells, Cultured ; Cicatrix ; metabolism ; pathology ; Female ; Fibroblasts ; cytology ; drug effects ; metabolism ; Humans ; Male ; Middle Aged ; Phenanthrenes ; pharmacology ; RNA, Messenger ; genetics ; Transforming Growth Factor beta1 ; genetics ; metabolism ; Young Adult

OBJECTIVETo study the response of rat bone marrow mesenchymal stem cells (MSCs) to a single period of mechanical strain and expression patterns of transforming growth factor-beta (TGF-beta) and insulin-like growth factor-II (IGF-II) after mechanical stretch.

METHODSBone marrow MSCs were isolated from SD rats and cultured in vitro. A four-point bending apparatus were used to perform a single period of mechanical strain (2000 microepsilon, 40 min) on MSCs. Cellular proliferation and alkaline phosphatase (ALP) activity of MSCs were examined and gene expression patterns of TGF-beta and IGF-II were detected by SYBR green quantitative real-time RT-PCR.

RESULTSCell proliferation, ALP activity and expression of TGF-beta and IGF-II were all significantly up-regulated in stretched MSCs when compared with their controls. The mRNA levels of TGF-beta and IGF-II got top increase immediately after mechanical loading and increased about 51.44 and 8.92 folds, respectively, when compared with control cells. Expression of TGF-beta and IGF-II decreased with time and returned to control level at 12 h after mechanical stimulus, despite of a small increase at 6 h.

CONCLUSIONThe mechanical stretch can promote MSCs proliferation, up-regulate its ALP activity and induce a time-dependent expression increase of TGF-beta and IGF-II which in turn result in osteogenic differentiation of MSCs. Mechanical stimulus is a key stimulator for osteogenic differentiation of MSCs and vital for bone formation in distraction osteogenesis.

Animals ; Bone Marrow Cells ; Cell Differentiation ; Cell Proliferation ; Insulin-Like Growth Factor II ; Mesenchymal Stromal Cells ; Osteogenesis ; Osteogenesis, Distraction ; RNA, Messenger ; Rats ; Rats, Sprague-Dawley ; Somatomedins ; Transforming Growth Factor beta

OBJECTIVETo study the clinicopathologic features, diagnosis and differential diagnosis of myositis ossificans (MO).

METHODSThe clinical features, radiologic results and pathologic findings of 15 cases of MO (including biopsy and surgical specimens) were analyzed. The hematoxylin and eosin sections were reviewed under light microscope. Immunohistochemical staining for S-100 protein, vimentin, desmin, actin and osteonectin was performed.

RESULTSThe age of the patients ranged from 12 to 46 years. The male-to-female ratio was 11:4. Thirteen cases were located in the parosteum of long bone or subperiosteal soft tissue. The remaining two cases occurred in iliac region and palm, respectively. Five patients had history of injury, while 2 patients had operation before. Four patients had no history of trauma and the remaining one had unknown clinical history. Histologically, zonation pattern was not conspicuous in 10 biopsy cases and 8 corresponding surgical specimens. On the other hand, zonation pattern was observed in 5 biopsy cases and 7 corresponding surgical specimens. Follow up revealed relapses in two patients. Immunohistochemical study showed various degree of positivity for vimentin, desmin, actin and osteonectin. S-100 protein was focally positive in 2 of the cases. The Ki-67 index varied from 1% to 10%.

CONCLUSIONCorrect diagnosis of MO relies on correlation of clinical features, radiologic examination and pathologic findings.

Adolescent ; Adult ; Biopsy ; Child ; Female ; Humans ; Male ; Middle Aged ; Myositis Ossificans ; diagnosis ; genetics ; pathology ; S100 Proteins ; genetics ; Vimentin ; X-Rays ; Young Adult