Objective To compare the clinical efficacy and safety of domestic orlistat and imported orlistat in Chinese overweight and obese patients. Methods In a randomized, double-blinded and positive-controlled study, 228 adults (BMI 24-< 40 kg/m~2) evaluated at seven research centers were randomized to receive domestic orlistat or imported orlistat 120 mg 3 times a day with an energy-controlled diet for 24 weeks. Results After 24 weeks, domestic orlistat treated patients got significant weight-loss (5.0±3.7) kg, which was comparable with that of imported orlistat treated patients (4.5±3.5) kg (P=0.3922).Compared with the findings before treatment, there was significant decrease of systolic blood pressure (4.4±11.5)mm Hg (1 mm Hg=0.133 kPa) and serum levels of TC (0.54±0.79) mmol/L and LDL-C (0.32±0.64) mmol/L in the domestic orlistat treated group(compared with levels of baseline, P< 0. 0001). There was no significant difference between the two groups in the changes of blood pressure and lipid levels. Both groups had similar adverse event profiles, most of which were mild and transient gastrointestinal events. There were no serious adverse events in beth groups. Conclusions Domestic orlistat combined with a light low-energy diet promoted significant weight loss, which was comparable with that of imported orlistat after 24 weeks of treatment. There was also improvement in blood pressure and serum levels of TC and LDL-C. Domestic orlistat was as effective and safe as imported orlistat in the treatment of obesity.

Bone morphogenetic protein 2(BMP-2) is a member of the of BMPs family, its osteoinductive capacity has already been demonstrated. We tried to express hBMP-2 in CHO cell. In this study, we inserted hBMP-2 cDNA into vector pCDNA3.1(+) to construct hBMP-2 eukaryotic expression vector pCDNA3.1(+)-hBMP-2. Recombinant Chinese hamster ovary (rCHO) cell line expressing high-level recombinant human bone morphogenetic protein 2(rhBMP-2) was constructed by co-transfecting the expression vectors pCDNA3.1(+)-hBMP-2 and plasmid pSV2-dhfr into dihydrofolate reductase (dhfr)-deficient CHO cells and the subsequent gene amplification in medium containing stepwise increments in methotrexate level such as 0.1 and 1 micromol/L. Western blot analyses showed a specific band of about 18 kD in reduced sample lane and a specific band of about 32 kD in non-reduced sample lane, this indicated that rCHO cells secret rhBMP-2 as a homodimeric glycoprotein form. Finally, we obtained a single clone cell strain expressing a high level (7.83 microg/24 h/10(6) cells) of rhBMP-2 tested by ELISA. Biological activity of rhBMP-2 was tested by the induction of alkaline phosphatase(ALP) activity in C2C12 cells. We treated C2C12 with different concentration of rhBMP-2 condition medium(CM) for 5d. The results showed that the rhBMP-2 could significantly increase the ALP activity of C2C12.
Alkaline Phosphatase ; biosynthesis ; Animals ; Blotting, Western ; Bone Morphogenetic Protein 2 ; biosynthesis ; chemistry ; isolation & purification ; pharmacology ; CHO Cells ; Cell Line ; Cricetinae ; Cricetulus ; Enzyme Induction ; drug effects ; Gene Expression ; Genetic Vectors ; genetics ; Humans ; Mice ; Recombinant Proteins ; biosynthesis ; chemistry ; isolation & purification ; pharmacology ; Solubility

BMP6 is a member of TGF-beta superfamily, represent more effective osteogenic activity. Two recombinant plasmids were constructed to expression rhBMP6 in mammalian cells, one contained the cDNA encoding the signal peptide, propeptide and mature peptide of human BMP6, wich was named pcDNA-BMP6, the other contained the recombinant DNA encoding the signal peptide, propeptide of human BMP2 and the mature peptide of BMP6, which was named pcDNA-BMP2/6. Transient expression in Cos7 cells demonstrated that the pcDNA-BMP2/6 produced more rhBMP6 than pcDNA-BMP6. For stable expression, the CHO-dhfr- cells were transfected with pcDNA-BMP2/6 and pSV2-dhfr, then screened by G418 and treated with MTX for targeting gene amplification. The partially purified rhBMP6 by heparin affinity chromatography was shown to possess bone induction activity tested by the induction of alkaline phosphatase activity in C2C12 cells.
Alkaline Phosphatase ; metabolism ; Animals ; Blotting, Western ; Bone Morphogenetic Protein 2 ; genetics ; Bone Morphogenetic Protein 6 ; genetics ; metabolism ; pharmacology ; CHO Cells ; COS Cells ; Cell Line ; Cercopithecus aethiops ; Cricetinae ; Cricetulus ; Gene Expression ; Humans ; Myoblasts ; cytology ; drug effects ; enzymology ; Protein Precursors ; genetics ; Protein Sorting Signals ; genetics ; Recombinant Fusion Proteins ; genetics ; metabolism ; pharmacology ; Reverse Transcriptase Polymerase Chain Reaction ; Transfection

AIM To establish the HPLC fingerprints of Yupingfeng Powder aqueous decoction and to determine the contents of nine constituents.METHODS The analysis of aqueous decoction was developed on a 30 ℃ thermostatic Hypersil ODS column (250 mm × 4.6 mm,5 μm),with the mobile phase comprising of acetonitrile-water flowing at 1.0 mL/min in a gradient elution manner,and the detection wavelength was set at 220 nm.RESULTS There were fifteen common peaks in the fingerprints of ten batches of samples,with the similarities of more than 0.95.Nine of them were identified as prim-O-glucosylcimifugin,calycosin-7-O-β-D-glucoside,cimifugin,4'-O-β-glucopyranosyl-5-O-methylvisamminol,psoralen,calycosin,sec-O-glucosylhamaudol,formononetin and atractylon,which showed good linear relationships within their own ranges (r ≥ 0.999 7),the average recoveries were 97.91％-99.81％ with the RSDs of 0.58％-1.27％.CONCLUSION This stable and reliable method can beused for the quality control of Yupingfeng Powder aqueous decoction.

BACKGROUNDBone morphogenetic protein-6 (BMP-6) is closely correlated with tumor differentiation and skeletal metastasis. Estrogen is considered as a stimulant for the initiation and promotion of breast cancer. Previous studies demonstrated that 17beta-estadiol (E2) can selectively increase the expression of BMP-6. This experiment is designed to detect the molecular mechanism of estrogen activating BMP-6 gene transcription in human estrogen receptor positive (ER+) breast cancer cell line MCF-7.

METHODSAfter the treatment of MCF-7 cells with E2 at different concentrations (10(-11) mol/L, 10(-9) mol/L, 10(-7) mol/L), the BMP-6 expression level was examined through real-time polymerase chain reaction. Through restriction enzyme digestion, human BMP-6 1.2 kb long promoter, BMP-6 0.7 kb long promoter was cloned into pGL-3 basic vector; after the treatment with 10(-7) mol/L E2, luciferase activities of the two promoters were detected. Site-directed mutagenesis was performed to obtain the mutant forms of estrogen response element half-site (1/2 ERE) element and Sp1 sites in the BMP-6 promoter, the activities of these mutant form promoters were detected following the methods mentioned above. Chromatin immunoprecipitation (ChIP) assay was also used to confirm the binding of estrogen receptor alpha (ERalpha) on BMP-6 promoter in the presence of E2.

RESULTSE2 dose dependently increased BMP-6 mRNA expression in human ER+ breast cancer cell line MCF-7. At a dose of 10(-7) mol/L E2, human BMP-6 1.2 kb promoter activity was increased by 90% compared with the control group treated with ethanol (P < 0.05). Both the 1/2 ERE response element mutant form and the Sp1 site mutant form of the BMP-6 promoter abolished the activation of the BMP-6 promoter's response to E2. Through ChIP assay, the binding of ERalpha on 1/2 ERE response element in BMP-6 promoter was further validated.

CONCLUSIONEstrogen induces BMP-6 expression in human ER+ breast cancer cell line MCF-7 through its receptor ERalpha binding on 1/2 ERE element in the BMP-6 promoter.

Bone Morphogenetic Protein 6 ; Bone Morphogenetic Proteins ; genetics ; Breast Neoplasms ; genetics ; Cell Line, Tumor ; Dose-Response Relationship, Drug ; Estradiol ; pharmacology ; Estrogen Receptor alpha ; physiology ; Female ; Humans ; Parathyroid Hormone-Related Protein ; secretion ; Promoter Regions, Genetic ; Transcriptional Activation ; drug effects

OBJECTIVE: To compare the results between auditory steady-state response (ASSR) and 40 Hz auditory event related potential (AERP), and explore the accuracy of hearing thresholds by using ASSR and AERP and the clinic forensic value. METHODS: Thirty seven ears were tested with pure-tone audiometer, 40Hz AERP and ASSR, respectively. All the volunteers in our study were awake during 40 Hz AERP test and ASSR test. RESULTS: Thresholds acquired with ASSR and 40Hz AERP test had a close correlativity and showed higher than those acquired with PTA test. There was no significant difference between the accuracy of ASSR and 40Hz AERP in estimating pure-tone thresholds. CONCLUSION After determining the correct value, ASSR can be used directly to evaluate hearing loss objectively.
Acoustic Stimulation ; Adult ; Audiometry, Evoked Response ; Audiometry, Pure-Tone/methods* ; Auditory Threshold ; Evaluation Studies as Topic ; Evoked Potentials, Auditory, Brain Stem/physiology* ; Female ; Hearing Loss, Sensorineural/physiopathology* ; Humans ; Male ; Middle Aged ; Predictive Value of Tests ; Sleep/physiology* ; Wakefulness ; Young Adult

OBJECTIVETo investigate the feasibility and efficacy of tubed pectoralis major myocutaneous flap in the reconstruction of circumferential defects following resection for locally advanced hypopharyngeal and cervical esophageal carcinoma.

METHODSFrom Dec. 2004 to Oct. 2008, 30 patients underwent immediate reconstruction by tubed pectoralis major myocutaneous flap for circumferential defects following resection of primary tumours. Among them, 22 were hypopharyngeal carcinoma, 7 were cervical esophageal carcinoma and one was recurrent laryngeal carcinoma involved the hypopharyngeal lumen. Five of 30 patients had received previous radiotherapy and three had failed in the previous surgical procedure. In this series, 12 patients had total pharyngolaryngectomy and 18 had total pharyngolaryngectomy and partial cervical esophagectomy.

RESULTSPostoperative pharyngocutaneous fistula formation occurred in 4 patients, 2 of them with previous radiotherapy and 2 with diabetes, and the fistulae healed later. Two patients developed anastomotic strictures at the upper junction, but they had good responses to dilatation treatment and had satisfactory oral intake. The postoperative follow-up time ranged from 8 to 56 months. Median follow-up was 18 months. One-year survival rate was 71.4% and three-year survival rate was 42.5%.

CONCLUSIONSThe tubed pectoralis major myocutaneous flap is a reliable procedure to reconstruct hypopharyngeal circumferential defects following resection of advanced hypopharyngeal and cervical esophageal carcinoma. This method may be the optimal choice for the reconstruction of hypopharyngeal circumferential defects following resection of recurrent carcinoma. The incidence of fistula and stenosis could be kept at an acceptable level.

Adult ; Aged ; Carcinoma, Squamous Cell ; surgery ; Esophagus ; pathology ; surgery ; Humans ; Hypopharyngeal Neoplasms ; surgery ; Hypopharynx ; surgery ; Male ; Middle Aged ; Neck ; surgery ; Pectoralis Muscles ; transplantation ; Reconstructive Surgical Procedures ; methods ; Surgical Flaps

OBJECTIVETo investigate the mechanisms of PTEN gene inactivation starting from DNA, mRNA and protein levels in ovarian cancers.

METHODSTumor tissue samples were obtained from 48 patients with epithelial ovarian cancers. Using four polymorphic markers (D10s541, D10s583, D10s1687 and D10s2491) within and flanking the PTEN gene located in chromosome 10q 23.3, polymerase chain reaction (PCR) and loss of heterozygosity (LOH) were introduced to examine LOH of PTEN gene; PCR-single strand conformation polymorphism (PCR-SSCP) was introduced to examine mutations of the fifth, sixth, seventh, and eighth exons of PTEN. Reverse transcriptase-polymerase chain reaction (RT-PCR) and immunohistochemistry (SP method) were applied to detect PTEN mRNA and PTEN protein expressions, respectively.

RESULTSLOH of PTEN gene was observed in 19 of 48 (39.6%) ovarian cancers. PTEN mutations were found only in 2 (4.2%) of the cases. Absence of PTEN mRNA expression was 18.8% (9 of 48). Immunostaining of 48 cancer samples revealed that 13 (27.1%) were PTEN immunostain negative. Of these 13 samples, only 2 (15.4%) had structural, biallelic inactivation by intragenic PTEN mutations and loss of the remaining wild-type allele; 7 (53.8%) showed evidence of LOH, 5 of these 7 samples showed deletion of PTEN mRNA expression, another 2 samples showed positive expression of PTEN mRNA; 4 (30.8%) tumors had neither PTEN gene mutation nor LOH but exhibited no PTEN protein expression, 2 of these 4 cases showed deletion of PTEN mRNA expression, another 2 showed positive expression of PTEN mRNA. For the cases of PTEN protein absent staining, the rate of LOH was 69.2% (9 of 13), higher than 28.6% (10 of 35) for the positive staining (P < 0.05).

CONCLUSIONSPTEN gene inactivation may contribute to epithelial ovarian carcinogenesis. There may be several mechanisms of PTEN gene inactivation in ovarian cancers. Protein expression deletions may be a significant mechanism.

Adult ; Aged ; Chromosomes, Human, Pair 10 ; Exons ; Female ; Gene Deletion ; Gene Expression Regulation, Neoplastic ; Genes, Tumor Suppressor ; Humans ; Loss of Heterozygosity ; Middle Aged ; Mutation ; Ovarian Neoplasms ; genetics ; metabolism ; PTEN Phosphohydrolase ; biosynthesis ; genetics ; Polymerase Chain Reaction ; Polymorphism, Single-Stranded Conformational ; RNA, Messenger ; biosynthesis ; genetics

OBJECTIVETo investigate the change of skeleton structure and masseter after mandibular angle osteotomy and its clinical significance in preoperative design.

METHODS3-dimensional CT cephalometry was performed before and after surgery in 18 cases of prominent mandibular angle.

RESULTSPre- and post-operative data were compared. Significant differences between the pre- and post-operative data were found in the mandibular angle, the distance between mandibular angle, length of ramus, ectropion angle of mandibular angle, triangle Go-Me-Go, the thickness, width and length of masseter.

CONCLUSIONSIt indicates the skeleton structure is changed and some kind of atrophy happens in the masseter after operation. So we suggest partial resection of masseter should be unnecessary for mandibular angle osteotomy.

Cephalometry ; methods ; Facial Bones ; diagnostic imaging ; Female ; Humans ; Imaging, Three-Dimensional ; Mandible ; diagnostic imaging ; Masseter Muscle ; diagnostic imaging ; Osteotomy ; Skull ; diagnostic imaging ; Tomography, X-Ray Computed ; Young Adult

OBJECTIVETo observe the effect and safety of supplying sodium chloride in the treatment of patients with severe heart failure.

METHODSConsecutive 51 hospitalized patients with severe heart failure and cardiac edema were included in this study. Normal diet (6 g NaCl/d) was supplied to all patients. On the basis of controlling fluid intake and treating related etiological factors as well as standard medications including furosemide for severe heart failure, patients with mild hyponatremia (serum sodium level 130 - 134 mmol/L) ate additional salted vegetables, patients with moderate hyponatremia (serum sodium level 125 - 129 mmol/L) and severe hyponatremia (serum sodium level < 125 mmol/L) ate additional salted vegetables and were received additionally intravenous 3%NaCl hypertonic saline infusion (10 ml/h) until reaching normal serum sodium level.

RESULTSOn admission, 37.25% (19/51) patients had hyponatremia. During the first two weeks hospitalization period, 88.24% (45/51) patients were treated with intravenous diuretics and total incidence of hyponatremia was 64.71% (33/51), mild hyponatremia was 50.98% (26/51), middle and severe hyponatremia was 13.73% (7/51); among them, hyponatremia lasted less than 3 d in 57.58% (19/33) patients and ≥ 3 d in 42.42% (14/33) patients. Heart failure exacerbation and hypernatremia were not observed in patients receiving additional sodium chloride therapy. Hospitalization time was similar among patients with different blood natrium levels [average (16 ± 12) d]. Fifty out of 51 (98%) patients discharged from the hospital with improved heart failure symptoms and signs.

CONCLUSIONSupplying additional sodium chloride could rapid correct hyponatremia in heart failure patients with or without intravenous diuretics therapy which might contribute to a favorable prognosis in hospitalized heart failure patients.

Adult ; Aged ; Aged, 80 and over ; Female ; Heart Failure ; drug therapy ; Humans ; Hyponatremia ; etiology ; prevention & control ; Male ; Middle Aged ; Prognosis ; Retrospective Studies ; Sodium Chloride ; administration & dosage ; adverse effects ; therapeutic use ; Sodium Chloride, Dietary