BACKGROUND:Estrogen/progestins replacement therapy prevents excess bone loss in postmenopausal women.Recently osteoprotegerin (OPG) has been identified in osteoblast and displayed to inhibit bone resorption.OBJECTIVE: To compare the action between 17β-estradiol (E2) and progesterone on OPG expression in cultured normal human osteoblast-like cells (hOB).DESIGN: A comparative investigation.SETTING: Institute of Metabolic Endocrinology, the Second Xiangya Hospital of Central South University.MATERIALS: α-MEM (Sigma Chemical Corp., St. Louis, MO, USA); Type Ⅳ collagenase (Sigma); Fetal bovine serum (Gibco-BRL Corp., Grand Island, NY, USA); Osteocalcin radioimmunoassay kit (DiaSorin Corp., Stillwater, MN, USA).METHODS: The experiments were carried out in the Institute of Metabolic Endocrinology, Second Xiangya Hospital of Central South University from January 2003 to March 2006. The osteoblasts were extracted from the cancelous bone of anterior superior iliac spine of normal people, then cultured. The hOB were treated with E2 and progesterone, and the expressions of OPG mRNA and OPG protein were determined by Northern blot analysis and enzyme-linked immunoabsorbent assay (ELISA) respectively.MAIN OUTCOME MEASURES: ①Characterization of human osteoblast-like cells; ②Effect of E2 and progesterone on OPG mRNA levels by Northern blot analysis; ③ Effect of E2 and progesterone on OPG protein levels in the conditioned medium by ELISA.RESULTS: ① Characterization of hOB in vitro The ALP levels in normal human osteoblasts were (74.3±4.7) U/g protein,and the detectable osteocalcin levels was (3.84±0.39) μg/L protein], which suggested that osteoblasts were the primary cell type found in our bone-derived cell cultures from donors. ② Effects of E2 and progesterone on the levels of OPG mRNA by Northern blot analysis: The OPG mRNA band was week in the control group [(12.3±3.5)%], treatment with 1 × 10-10, 1 ×10-9 1 ×10-8 mol/L E2 caused an increase in the levels of OPG mRNA. The expression of OPG mRNA in the 1×10-8 mol/L E2 group was gradually increased at 12, 24 and 48 hours. Progesterone had no influence on OPG mRNA expression. ③ Effects of E2 and progesterone on OPG protein production in conditioned medium determined with ELISA:ELISA revealed that treatment with 1 ×10-10, 1 ×10-9, 1 ×10-8 mol/L E2 induced obvious increase in the levels of OPG protein in cells media as compared with that in the control group [(1.27±0.26), (2.34±0.35), (3.62±0.23), (0.64±0.14)μg/L, P ＜ 0.01]. In the presence of 1×10-8 mol/L E2, OPG protein production in cells media at 12, 24 and 48 hours were significantly higher than that in the control group [(1.30±0.30), (3.07±0.14), (3.50±0.33), (0.62±0.12) μg/L, P ＜ 0.01]. 1 × 10-10, 1 ×10-9 1 × 10-8 mol/L progesterone had no influence on the OPG protein production after 12-24 hours (P ＞ 0.05).CONCLUSION: The different regulation of OPG production in osteoblasts by E2 and progesterone may contribute to the mechanisms by which estrogen or progestins exerts its different action on bone resorption.

OBJECTIVETo identify the genetic alteration in human keloid.

METHODSComparative genomic hybridization was applied in 6 cases of keloid to investigate the genomic imbalance (the gain or loss of genetic material).

RESULTSThe study showed that the loss of chromosome DNA copies included chromosome, 1,7,9,13,16,17,18,19,20,22. Among them, the frequently detected chromosome loss was chromosome 1 p(66.7%), 16 (83.3%), 20 (83.3%) and 22 (83.3%). The minimum overlapping regions were 1 pter-32.2,16p13.2p11.l,20q11.1-q13.2 and 16p13.2-p11.1. Frequent gain of DNA copy numbers was not found in the special regions.

CONCLUSIONSThe mapping of DNA copy variation frequency in keloid showed that there may be inhibitory genes in chromosomes 1p,16,20,22. The loss of these genes may be involved in the development and progress of keloid.

Adult ; Chromosome Aberrations ; Chromosomes, Human, Pair 1 ; genetics ; Comparative Genomic Hybridization ; DNA Probes ; Female ; Humans ; Keloid ; genetics ; Male ; Middle Aged ; Young Adult

Objective To evaluate the efficacy of tibial bone-skin flap grafts in the management of se- vere traumatic osteomyelitis complicated with bone and skin defect in leg to avoid amputation.Methods From March 1998 to Aug.2004,12 cases of the traumatic osteomyelitis complicated with bone and skin defect in leg were treated with vascularized tibial bone-skin flap graft.The longest flap was 17cm,widethest 10cm, The longest bone flap was 12cm.They were followed up for 0.6 to 5 years.Results All the tibial bone-skin flaps survived completely,2 cases of osteomyelitis recurred.The followed-up,from 0.5 to five years,showed good bone union in all cases,averageing 15 weeks.The infection was under control.The leg function and con- tour were satisfactory.Conclusion The tibial bone-skin flap has the advantages of having distinguished sign of anatomy,highly vascularized,easy to obtain,simply and flexible procedure,improving circulation,short- ens hospitalization and suitable for treatment of traumatic osteomyelitis complicated with bone and skin defect in leg.

To investigate the metabolic rate and metabolites of 9-dehydro-17-dehydro-andrographolide, which is the main active ingredient in Xiyanping injection, by using the in vitro rat liver microsome incubation system. 9-dehydro-17-dehydro-andrographolide was incubated together with liver microsome mixed with NADPH. Its metabolic rate was studied by determining its residual concentrations with the UHPLC-MS/MS method; Its metabolites were identified by the UPLC-TOF-MS(E) method. The results showed that 9-dehydro-17-dehydro-andrographolide was metabolized faster than rat liver microsomes mixed with coenzymes, with t½ and CL of (19.7 ± 0.5) min and (35.1 ± 0.8) mL x min(-1) x g(-1) (protein), respectively. Based on the high resolution mass spectrum data and information from literatures, altogether nine metabolites of 9-dehydro-17-dehydro-andrographolide were identified in the incubation system, particularly hydroxylated and dehydrogenized products. The results of identification would provide a basis for screening out more active andrographolide derivatives.
Animals ; Chromatography, High Pressure Liquid ; Diterpenes ; chemistry ; metabolism ; Drugs, Chinese Herbal ; chemistry ; metabolism ; Microsomes, Liver ; chemistry ; metabolism ; Molecular Structure ; Rats ; Tandem Mass Spectrometry

Objective To investigate the efficacy and safety of CT-guided radiofrequency ablation (RFA) in treating lung tumors. Methods A total of 33 patients with lung cancer (35 lesions in total), who were admitted to authors’ hospital during the period from May 2007 to August 2013 to receive treatment, were enrolled in this study. RFA was carried out in all patients. After RFA the patients were followed up regularly (once every 3 months) to evaluate the therapeutic efficacy and the adverse reaction. The deadline for the following-up was November 2013, or to the time when tumor progression occurred. Results Of the total 34 lesions in 32 patients who had received RFA and had complete follow-up data, the one-year local control rate was 85.3%. The average one-year progression-free survival rate was 75.0%, among them 15 cases with primary lung cancer had a mean one-year progression-free survival rate of 80.0% and 17 cases with metastatic lung cancer had a mean one-year progression-free survival rate of 70.6%. The overall median progression-free survival (PFS) was(18.0±1.3) months. No obvious correlation existed between PFS and age, sex, tumor size, pathological type, clinical stage (P<0.05). The main adverse reactions of RFA were pain, hydrothorax and pneumothorax; no serious life-threatening complications occurred. Conclusion RFA is a safe, effective and minimally-invasive treatment for lung cancer, regardless of early stage or late stage of the tumor.

Objective To evaluate the safety and clinical efficacy of CT-guided 125I seeds interstitial implantation in treating the refractory liver cancers that show poor response to commonly used therapies. Methods A total of 40 patients with refractory clinically or pathologically-proved liver cancer were enrolled in this study, the diseases included primary liver cancer (n = 27, with coexisting portal vein cancerous thrombus in 2) and metastatic liver cancer (n = 13). CT-guided 125I seeds interstitial implantation was performed in all patients. Preoperative plan of seeds implantation protocol was formulated by using the treatment plan system (TPS); the 125I seed activity was 0.6 -0.8 mCi and the peripheral matching dose (MPD) was 100 -140 Gy. The procedure of 125I seeds interstitial implantation was performed under local anesthesia in all patients. By using percutaneous trans-hepatic puncturing and single-or multiple-needle technique, the 125I seeds were implanted along a line parallel to the long axis of the tumor and/or tumorous thrombus with an interval of 0.5 -1.0 cm. The short-term efficacy was evaluated by modified response evaluation criteria in solid tumors (mRECIST), and the median time to tumor progression (mTTP) and the median overall survival time (mOS) were calculated by Kaplan and Meier method. Results The technical success rate was 100%. The diameter of the tumor was 1.5 -12.0 cm (mean 4.0 cm), and a total of 1 748 125I seeds were implanted in 40 patients (mean 44 seeds per patient). The short-term effective rate was 37.5%(n = 15), including complete remission in 8 cases and partial remission in 7 cases, the stable disease was seen in 15 cases (37.5%), and the disease control rate was 75%. The mTTP was 7.0 months (95%CI:4.524-9.476 months), while mOS was 10 months (95%CI: 6.901 -13.099 months). The procedure-related adverse reactions included small amount of subcapsular hemorrhage (n =2, 5%), intrahepatic migration of 125I seeds (n=2, 5%), pain at liver area (n=1, 2.5%); and no special treatment was needed in these patients. One patient developed high fever with chills 3 hours after the procedure, which was relieved after symptomatic and antipyretic treatment. Conclusion For the treatment of refractory liver cancers, CT-guided 125I seeds permanent interstitial implantation, used as a remedial therapy, is safe and effective. This technique is worth popularizing in clinical practice.

Objective To investigate the regularity of abdominal pain and its influence factors in patients with primary hepatic carcinoma (PHC) after receiving transcatheter arterial chemoembolization (TACE).Methods A total of 121 patients with PHC were enrolled in this study.All patients were admitted to the Department of Interventional Radiology of Zhejiang Provincial Cancer Hospital from December 2012 to June 2013,and all patients were suffered from PHC.The occurrence,duration and severity of the abdominal pain as well as the used dosage of morphine within 48 hours after TACE were documented.The results were statistically analyzed.Results A total of 96 patients (96/121,78.5％) complained of different degrees of abdominal pain after interventional therapy,and 72 patients (72/121,59.5％) showed moderate to severe pain,with the VAS score being more than 4 points.The average dosage of morphine used each time for one patient was 19.7 mg.Statistical analysis indicated that these patients were more prone to develop abdominal pain after TACE if they carried more than one of the following risk factors:age ≥60 years (when compared with patients ＜60 years,OR:0.307,P=0.008),preoperative ECOG score ＞2 (when compared with a ECOG score of 0-1,OR:0.195,P=0.006),the distance between tumor and liver capsule ＞1 cm (when compared with the distance ≤ 1 cm,OR:0.296,P=0.007),the use of THP in performing chemoembolization (when compared with other chemotherapeutic drugs,OR:0.232,P＜0.003 4).Conclusion After TACE abdominal pain is a high-frequency event.The independent factors affecting the occurrence of abdominal pain are age＜60 years,preoperative ECOG score ＞2,tumor located close to liver capsule,and the use of THP-lipiodol mixture as embolic agent.Therefore,for patients carrying moderate-high risk of abdominal pain,routine use of analgesics before TACE as well as within 12 hours after TACE to prevent the occurrence of abdominal pain is quite necessary.

OBJECTIVETo identify the genetic alterations in nonsyndromic cleft lip and palate (NSCLP).

METHODSComparative genomic hybridization was applied to investigate the genomic imbalance (the gain or loss of genetic material) in 7 cases of NSCLP.

RESULTSIt showed that the loss of chromosome DNA copies happened in chromosome 6, 7, 10, 13, 14, 16, 20, 22 and the gain of chromosome DNA copies happened in chromosome 5, 15, 18, 19. Conclusions 13q had a high frequency (71.4%) of chromosome loss.

CONCLUSIONSAbnormal chromosome DNA copies happen in all the patients with NSCLP. Most of the patients have chromosome DNA copies loss. It suggests that loss of inhibitory gene may be related to the NSCLP. The related inhibitory gene may locate in 13q.

Adolescent ; Adult ; Child ; Child, Preschool ; China ; ethnology ; Cleft Lip ; genetics ; Cleft Palate ; genetics ; Comparative Genomic Hybridization ; DNA ; Genetic Variation ; Genotype ; Humans ; Infant ; Mutation ; Phenotype ; Young Adult

OBJECTIVETo study the mutation in RH120480 fragment of RUNX3 gene among the Chinese patients with keloid.

METHODS20 samples of keloids were collected with each patient's venous blood sample as normal control group. The genomic DNA was extracted from each sample. RH120480 fragment of RUNX3 gene was amplified by Polymerase Chain Reaction (PCR). The amplification products were analyzed by denaturing high-performance liquid chromatography (DHPLC). Some fragments were sequenced directly and then compared with the GenBank data.

RESULTSBy DHPLC, the results of all the blood samples showed single chromatographic peak indicating homoduplexes, meanwhile the results of keloid tissue samples showed double peak indicating heteroduplexes. Through gene sequencing, 19 cases showed gene mutation among the 20 samples of keloid. The mutation incidence was 95%. Two mutation sites were detected including base A absence in 96th sites and base C insert in 279th sites. The base A absence rate was 90% (18/20) in keloid group, and 10% (2/20) in control group. The base C insert mutation rate was 95% (19/20) in keloid group, and 0% (0/20) in control group. There was significant difference in the mutation rate between two groups on the two mutation sites.

CONCLUSIONSThere is a strong correlation between the RH120480 fragment of RUNX3 gene mutation and Keloid. RUNX3 gene could be possibly a scar suppressor gene (SSG).

Adolescent ; Adult ; Core Binding Factor Alpha 3 Subunit ; genetics ; DNA ; genetics ; Female ; Humans ; Keloid ; genetics ; Male ; Middle Aged ; Mutation ; Young Adult

OBJECTIVEThe aim of this study was to investigate the loss of heterozygosity (LOH) on chromosome 1 pter-36.21 of keloid in order to locate the deletion areas probably harboring scar suppressor genes.

METHODSUsing polymerase chain reaction ( PCR )-denaturing polyacrylamide gel electrophoresis, 25 samples of keloid tissues and peripheral blood were analyzed.

RESULTS15 out of 25 samples of keloid tissues exhibited LOH in at least one microsatellite locus. There were deletions at more than one locus of one keloid tissue. No MSI was found. The frequency of LOH was remarkably higher in the keloid tissues (n = 25, 15, 60%) than in the normal control samples (n = 25, 1, 4%). The frequency of LOH in D1S243, D1S468, D1S507 and D1S199 was as following: (n= 25, 7, 28%), (n =25, 10, 40%), (n = 25, 13, 52%) and (n= 25, 3, 12%). The frequency of LOH in D1S243, D1S468, D1S507 were statistically significant.

CONCLUSIONThe most common LOH occurred at D1S243-D1S468-D1S507 might imply the existence of potential tumor suppressor gene of a subset of keloid , while MSI on 1 pter-36.21 may not a crucial event.

Adolescent ; Adult ; Chromosome Mapping ; Chromosomes, Human, Pair 1 ; Female ; Humans ; Keloid ; genetics ; Loss of Heterozygosity ; Male ; Middle Aged ; Young Adult