Thirty-five patients with ketosis-prone diabetes (KPD) and 38 patients with newly-diagonosed common type 2 diabetes mellitus (DM) were treated with insulin for 3 months.The results showed that the existing insulin resistance in patients with KPD was mild as compared with common type 2 DM, and the recovery of insulin sensitivity was less marked in patients with KPD after blood glucose was well controlled.

To evaluate the clinical efficacy and safety of Qinghouyan lozenge in the treatment of acute pharyngitis due to Lung-heat and Yin-deficiency, and compare with Qinghouyan oral Liquid. Totally 144 subjects were enrolled and randomly divided into two groups (72 in the test group and 72 in the control group). The participants in the test group were given Qinghouyan lozenge for 5 days, and those in the control group were given Qinghouyan oral Liquid for 5 days. The effectiveness evaluation indexes were pharyngalgia/odynophagia disappearance rate, overall efficacy of TCM syndromes, TCM syndrome scores, and single syndrome and sign disappearance rate. During the test, the safety was evaluated by vital sign, lab examination indexes and adverse events. The results for the full analysis set showed that the couth disappearance rate, the incidence rate of TCM syndromes, and the throat/uvula congestion disappearance rate of the test group were higher than that of the control group (P < 0.05), with significant differences in the changes in syndrome scores between the two groups (P < 0.05). Altogether 3 adverse events were observed in the test group while 6 adverse events in the control group, without significant differences in the adverse event rate between the two groups (P < 0.05), serious abnormal laboratory examinations and vital signs. In conclusion, Qinghouyan lozenge has better efficacy in treatment of acute pharyngitis due to Lung-heat and Yin-deficiency than Qinghouyan oral liquid, with good safety.
Acute Disease ; Double-Blind Method ; Humans ; Medicine, Chinese Traditional ; Pharyngitis ; drug therapy

BACKGROUNDJaw osteonecrosis possibly associated with the administration of bisphosphonates is expected to be treated with a non-pharmacologic approach. This study aimed to determine whether noninvasive, mechanically mediated vibration would inhibit the decline in bone mineral density (BMD) that follows menopause, enhance the BMD of the lumbar and femoral neck, and reduce chronic back pain in postmenopausal women with osteoporosis.

METHODSA total of 116 postmenopausal women with osteoporosis participated in this study, and they were divided into groups A (66 patients) and B (50). Group A received vibration treatment (Subjects vertically stand on the vibration platform, with a vibration frequency of 30 Hz, amplitude of 5 mm; they received the treatment five times per week, ten minutes each time and totally for six months), whereas women of group B served as controls without any treatment. L2 - 4 BMD, bilateral femoral neck BMD, and body mass index (BMI) were recorded before the treatment or at the third and sixth months of the treatment respectively. After the ending of the treatment, the change of BMD in each group was compared and analyzed. Chronic back pain was evaluated by visual analogue scale (VAS) at baseline and the third and sixth months of the treatment.

RESULTSOf the 116 women, 94 including 51 women from group A ((61.23 +/- 8.20) years) and 43 women from group B ((63.73 +/- 5.45) years), completed the study. There were no significant differences in baseline characteristics including age, BMI, menopausal years, lumbar BMD, femoral neck BMD, and VAS between the two groups. The lumbar BMD of the 51 women in group A increased by 1.3% (P = 0.034) after vibration treatment for 3 months and by 4.3% at the sixth month (P = 0.000). The lumbar BMD in group B was decreased at the third month, but there was not statistical significance (P > 0.05). At the sixth month, it was decreased by 1.9% (P < 0.05). The femoral neck BMD of the 51 women in group A was slightly increased after vibration treatment for 3 months, but without statistical significance (P > 0.05). At the sixth month, the BMD was increased by 3.2% (P < 0.05). In group B, the BMD was not decreased significantly (P = 0.185) at the third month, but decreased significantly at the sixth month (1.7%) (P < 0.05) compared with the baseline. Chronic back pain (VAS) reduced more significantly in group A at the third and the sixth months (P < 0.05) after vibration therapy in comparison with the baseline. The BMI was not significantly changed in the two groups during the period of follow-up.

CONCLUSIONSVibration therapy appears to be useful in reducing chronic back pain and increasing the femoral neck and lumbar BMD in postmenopausal women with osteoporosis.

Aged ; Back Pain ; prevention & control ; Bone Density ; Female ; Femur Neck ; Humans ; Lumbar Vertebrae ; Middle Aged ; Osteoporosis, Postmenopausal ; therapy ; Vibration ; therapeutic use

OBJECTIVETo investigate the effect of antisense vascular endothelial growth factor (VEGF)(121) cDNA transfection on the growth of K562 cells in nude mice.

METHODSK562 cells transfected with the antisense (AS) or sense (S) VEGF(121) cDNA, and the vector (V, pcDNA3) alone were transplanted subcutaneously into nude mice and the growth of the transfected cells in vivo was investigated. The effects of transfected K562 cells on human bone marrow endothelial cells (BMEC) were analyzed by MTT assay, the microvessel density (MVD) in tumor mass by vWF immunohistochemistry stain.

RESULTSK562/V tumor grew more slowly [(207.5 +/- 192.9) mm(3) vs (445.0 +/- 150.9) mm(3), P < 0.05] and K562/S tumor more rapidly than K562/V tumor did [(1 174.6 +/- 508.7)/mm(3) vs (445.0 +/- 150.9) mm(3), P < 0.01]. K562/S cell culture supernatant was more strongly in promoting the proliferation of BMEC than K562/V supernatant did, but K562/AS supernatant resulted in a marked decrease of the promoting effect as compared with K562/V's. The MVDs in K562/AS, K562/S, and K562/V tumors were [(11.0 +/- 7.6)/0.72 mm(2) vs (50.8 +/- 11.7)/0.72 mm(2) vs (18.9 +/- 7.0)/0.72 mm(2)], respectively.

CONCLUSIONSAntisense VEGF(121) cDNA transfected K562 cells show growth retardation in transplanted nude mice, decrease of tumor MVD, and decrease of promoting BMEC proliferation capacity.

Animals ; Bone Marrow Cells ; cytology ; drug effects ; Cell Division ; genetics ; physiology ; Culture Media, Conditioned ; pharmacology ; DNA, Antisense ; genetics ; DNA, Complementary ; genetics ; Endothelial Growth Factors ; genetics ; physiology ; Endothelium, Vascular ; cytology ; drug effects ; Female ; Humans ; K562 Cells ; Lymphokines ; genetics ; physiology ; Mice ; Mice, Inbred BALB C ; Mice, Nude ; Neoplasm Transplantation ; Neoplasms, Experimental ; blood supply ; genetics ; pathology ; Neovascularization, Pathologic ; genetics ; physiopathology ; Transfection ; Vascular Endothelial Growth Factor A ; Vascular Endothelial Growth Factors

OBJECTIVETo investigate the polymorphism of human cytomegalovirus (HCMV) UL150 open reading frame (ORF) in low-passaged clinical isolates, and to study the relationship between the polymorphism and different pathogenesis of congenital HCMV infection.

METHODSPCR was performed to amplify the entire HCMV UL150 ORF region of 29 clinical isolates, which had been proven containing detectable HCMV-DNA using fluorescence quantitative PCR. PCR amplification products were sequenced directly, and the data were analyzed.

RESULTSTotally 25 among 29 isolates were amplified, and 18 isolates were sequenced successfully. HCMV UL150 ORF sequences derived from congenitally infected infants were high variability. The UL150 ORF in all 18 clinical isolates shifted backward by 8 nucleotides leading to frame-shift, and contained a single nucleotide deletion at nucleotide position 226 compared with that of Toledo strain. The nucleotide diversity was 0.1% to 6.8% and the amino acid diversity was 0.2% to 19.2% related to Toledo strain. However, the nucleotide diversity was 0.1% to 6.4% and amino acid diversity was 0.2% to 8.3% by compared with Merlin strain. Compared with Toledo, 4 new cysteine residues and 13 additional posttranslational modification sites were observed in UL150 putative proteins of clinical isolates. Moreover, the UL150 putative protein contained an additional transmembrane helix at position of 4-17 amino acid related to Toledo.

CONCLUSIONHCMV UL150 ORF and deduced amino acid sequences of clinical strains are hypervariability. No obvious linkage between the polymorphism and different pathogenesis of congenital HCMV infection is found.

Amino Acid Sequence ; Base Sequence ; Cytomegalovirus ; genetics ; isolation & purification ; Cytomegalovirus Infections ; congenital ; virology ; DNA, Viral ; genetics ; Genes, Viral ; Genetic Variation ; Humans ; Infant ; Infant, Newborn ; Molecular Sequence Data ; Open Reading Frames ; Polymorphism, Single Nucleotide ; Sequence Homology, Amino Acid ; Viral Proteins ; chemistry ; genetics

According to our previous experiments, Ph(+) chronic myeloid leukemia (CML) cell line K562 cells have defects in beta 1 integrin activation. In order to search the same regularity in Ph(+) CML bone marrow cells, bone marrow mononuclear cells (BMMNC) from 12 cases of Ph(+) CML and 10 cases of normal individuals were studied. Their expression rate of 9EG7 epitope on beta1 integrin post treatment by 8A2 or GM- or G-CSF and cell adhesion ability with soluble fibronectin (FN) were evaluated by flow cytometry; in addition, the effects of CGP57148B, a highly specific ABL tyrosine kinase inhibitor, were observed. Our results showed that 9EG7 expression rate and FN binding rate were very low in all the inactivated cells. The parameter increased markedly post 8A2 activation in both NBMMNCs and CMLBMMNCs, but the degree of increase in CMLBMMNCs was significantly lower than that in NBMMNCs; GM-CSF or G-CSF could significantly increase the parameters in NBMMNCs while had no effects on that in CMLBMMNCs. CGP57148B could increase the beta1 integrin activation potential of CMLBMMNCs but had no effects on that of NBMMNCs. The results indicate that decreased activation potential of beta1 integrin in CMLBMMNCs is the major cause of adhesion defects of Ph(+) CML cells; beta1 integrin functional insufficiency in CMLBMMNCs could not be directly reversed by ABL tyrosine kinase inhibitor CGP57148B.

Host genetic factors, such as human leukocyte antigen (HLA) alleles, are important in Human immunod-eficiency virus (HIV) infection and its progression to AIDS. HLA class I genes, especially highly polymorphicHLA-B genes, are involved in the activation of HLA-restricted cytotoxic T lymphocytes (CTLs) against HIV, andthus control susceptibility to or protect against this virus. The present study was aimed to determine the distributionof HLA-B alleles in the Chinese Uygur ethnic group and its association with HIV infection. One hundred ten healthycontrol (HIV negative) and 128 HIV positive Chinese Xinjiang Uygur ethnic individuals were used in this study.HLA typing for B allele was performed by polymerase chain reaction (PCR) with sequence-specific primers (SSP).Hardy-Weinberg equilibrium was calculated using POPGENE software for the healthy control group. The HLA-Bfrequency of each allele was compared between the patients and the controls using the chi-square test. In HIV-1-pos-itive group, gene frequency of allele B * 4901 was significantly higher compared to the healthy control subjects (P=0.02, OR=3.06, 95%CI=1.16～8.10 forB*4901). In contrast, the gene frequency of B * 40 in healthy controlswas significantly higher than in the HIV-positive patients (P=0.02, OR=0.39, 95%CI=0.07～0. 92 for B* 40).In this study, HLA allele B * 4901 may be associated with increased susceptibility to HIV-1 infection, whereas the B* 40 allele may be associated with resistance to H HIV-1 infection.

OBJECTIVETo investigate the expression profiles of myocardin gene during the differentiation of bone marrow-derived mesenchymal stem cell to smooth muscle cells in the conditional medium combined with a high concentration of fetal bovine serum (FBS).

METHODSMarrow-derived mesenchymal stem cells were isolated and purified from mouse femoral bone and shinbones using differential adherent methods. Cells at the third passage were induced by 20% FBS in conditioned medium, conditioned medium alone, 20% FBS or 10% FBS alone respectively. Mouse aortic smooth muscle cells were cultured as the positive control. Levels of mRNA and protein expression of myocardin and several smooth muscle cells marker genes were determined by immunofluorescence, RT-PCR and Western blot before and 3, 7, 10, 14 d after the induction. The presence of smooth muscle myofilaments was detected by using transmission electron microscope.

RESULTSNaive bone marrow-derived mesenchymal stem cells displayed multiple morphological forms including fusiform, polygon, oval, and micro-spherical, as compared to the single macro-spindle form after the induction. Typical appearance of peak valley was displayed on the 21st day after induction. At the same time, the expression of smooth muscle marker genes was reinforced along with an up-regulation of myocardin expression. Immunofluorescence study showed that the cells expressing myocardin and smooth muscle marker genes such as alpha-SMA and SM-MHC increased. Fluorescence domain of myocardin translocated from cytoplasm to nucleus and the amounts of double positive cells for myocardin with alpha-SMA or SM-MHC also increased. RT-PCR confirmed that the mRNA expression of myocardin increased gradually and remained stabilized after achieving its peak on the 7th day after induction. The expression of smooth muscle marker genes, alpha-SMA and SM22alpha, remained stable on the 10th day of induction. It was also confirmed by Western blot that the protein expression of both myocardin and alpha-SMA were markedly increased during the induction. Finally, transmission electron microscopy revealed the presence of myofilament on the 21st day after induction.

CONCLUSIONSBone marrow-derived mesenchymal stem cells can be effectively induced into smooth muscle-like cells by conditioned medium combined with 20% FBS. Myocardin plays an important role in the differentiation process of bone marrow-derived mesenchymal stem cells to the peripheral smooth muscle cells.

Animals ; Bone Marrow Cells ; cytology ; physiology ; Cattle ; Cell Differentiation ; physiology ; Male ; Mesenchymal Stromal Cells ; cytology ; metabolism ; Mice ; Muscle, Smooth, Vascular ; cytology ; Myocytes, Smooth Muscle ; physiology ; Nuclear Proteins ; genetics ; metabolism ; physiology ; Reverse Transcriptase Polymerase Chain Reaction ; Trans-Activators ; genetics ; metabolism ; physiology ; Up-Regulation

Objective To construct the small interfering RNA (siRNA) targeting heparanase gene and its expressing vector,and to observe its interference effect on the expression of heparanase gene and inhibitory effect on the invasive potential of human malignant melanoma A375 cells.Methods Three siRNAs were designed.The recombinant plasmid pRNATU6.1/heparanase-siRNA was designed and constructed. A375 cells were cultured,and transfected with pRNATU6.1/heparanase-siRNA.The cells treated with lipo- fectamine or Opti-MEM served as the controls.Real-time fluorescence quantitative PCR and Western blot were performed to evaluate the expression of heparanase RNA and protein in these treated A375 cells.The in vitro invasive potential of treated A375 cells was assessed by Matrigel gel assay.Results The siRNA targeting heparanase gene was successfully cloned to the eukaryotic expressing vector pRNATU6.1.The expression levels of both heparanase RNA and protein decreased significantly in siRNA-transfected A375 cells than those in the control cells.The in vitro invasive potential of siRNA-transfected cells was also signifi- cantly inhibited as compared with that of the control cells (P

Vascular endothelial growth factor (VEGF) is a specific mitogen for vascular endothelial cells. It has been associated with angiogenesis, growth, metastasis and poor prognosis in solid tumors. Lately, it has been known that VEGF expression is higher in bone marrow from chronic myeloid leukemia (CML) patients than that in normal subjects. However, it is not clarified that the effect of VEGF on the abnormal proliferation of CML cells. In order to explore the effect of autocrine VEGF on CML cells, K562 cells were transfected with the VEGF(121) cDNA sense vector (K562/S) or with the VEGF(121) cDNA antisense vector (K562/As). K562 cells were transfected with the pcDNA(3) vector (K562/V) as the control. Cell proliferation was determined by MTT and colony forming assay in vitro. Flow cytometric Annexin-V-FITC/PI dual labeling technique was performed to observe the effect of VEGF(121) cDNA transfection on apoptosis of K562 cells. Results indicated that K562/S transfectants exhibited a 3-fold increase in VEGF secretion, and K562/As transfectants exhibited a 49% reduction in VEGF secretion. K562/As showed a reduced growth rate and colony forming efficiency as compared to K562/V. K562/S showed an increasing growth rate and colony forming efficiency as compared to K562/V. K562/As had more apoptotic cells than K562/V and K562/S in the same culture condition. These data suggest that VEGF plays an important role in the abnormal proliferation and apoptosis in CML cells through an autocrine mechanism.