OBJCETIVETo investigate the method of separation of culture of bone microvascular endothelial cells (BMECs) of human femoral head in vitro.

METHODSFrom October 2013 to January 2014,15 femoral heads without pathologic change from patients resected during hip replacement were selected involving 2 males and 13 females with a mean age of 71.2 years old ranging from 38 to 92. Cancellous bone in femoral head was bited into broken bone grain and transfered into medium in aseptic contidion. Cells were isolated by the methods of enzymic digestion and density gradient centrifugation,purified by differiential attachment. The characteristics of cells was observed by inverted microscope. vWF and CD31 immunofluorescence analysis was applied for identification of cells.

RESULTSThe number of cells was positively correlated with patients' age after 24 hours in primary culture. The older patients had the less cells numbered. After 4 to 5 days' culture, primary cells appeared short spindle,polygon shaped and cobblestone-like morphology. After 7 to 10 days' culture, primary cells proliferated densely, became fusion, arranged in swirl, and contact inhibition appeared significantly. Immunofluorescence staining revealed the cells were 100% positive for vWF and CD31, and it showed that the cultured cells were BMECs.

CONCLUSIONIt was a simple, steady, effective method with good reproducibility, by which highly purified human BMECs can be obtained.

Adult ; Aged ; Aged, 80 and over ; Cell Culture Techniques ; Cell Proliferation ; Cell Separation ; methods ; Cells, Cultured ; Endothelial Cells ; cytology ; Female ; Femur Head ; blood supply ; Humans ; Male ; Microvessels ; cytology ; Middle Aged

By studying the relationship between fingertip temperature changes and arterial function during vascular reactivity test, we established a new non-invasive method for detecting vascular function, in order to provide an assistance for early diagnosis and prevention of cardiovascular diseases. We customized three modules respectively for blood occlusion, measurement of finger temperature and blood oxygen acquisition, and then we established the hardware of data acquisition system. And the software was programmed with Labview. Healthy subjects [group A, n = 24, (44.6 ± 9.0) years] and subjects with cardiovascular diseases [group B, n = 33, (57.2 ± 9.9) years)] were chosen for the study. Subject's finger temperature, blood oxygen and occlusion pressure of block side during and after unilateral arm brachial artery occlusion were recorded, as well as some other regular physiological indexes. By time-domain analysis, we extracted 12 parameters from fingertip temperature signal, including the initial temperature (Ti), temperature rebound (TR), the time of the temperature recovering to initial status (RIt) and other parameters from the finger temperature signal. We in the experiment also measured other regular physiological body mass index (BMI), systolic blood pressure (SBP), diastiolic blood pressure (DBP) and so on. Results showed that 8 parameters difference between the two group of data were significant. based on the statistical results. A discriminant function of vascular function status was established afterwards. We found in the study that the changes of finger temperature during unilateral arms brachial artery occlusion and open were closely related to vascular function. We hope that the method presented in this article could lay a foundation of early detection of vascular function.
Adult ; Aged ; Arteries ; physiopathology ; Blood Pressure ; Cardiovascular Diseases ; diagnosis ; Fingers ; Humans ; Middle Aged ; Skin Temperature

Objective To observe the change of capillary pericapillary cells in rats with myocardial infarction and the influence of supplementing qi and activating blood circulation herbs, and explore its mechanism of improving myocardial perfusion. Methods The rat model was established by ligaturing the left anterior descending coronary artery. On the base of ECG evaluation, successfully modeled rats were randomly divided into the model group, group treated with supplementing qi and activating blood circulation Chinese medicine (activating blood and supplementing qi group), group treated with Perindopril (Perindopril group), group treated with Tongxinluo Capsules (Tongxinluo group). The sham-operation group was taken as the control. There were totally 5 groups. The model group and the sham-operation group were treated with normal saline. The changes of myocardial capillary density (MCD) and number of pericapillary cells on the 7th, 28th day after medicinal administration were observed. Results On the 7th and 28th day, the MCD decreased significantly and the number of capillary pericapillary cells increased significantly in the model group compared with the sham-operation group (P<0.01). The MCD increased significantly in the activating blood and supplementing qi group, Perindopril group and Tongxinluo group compared with the model group (P<0.01). The number of pericapillary cells decreased significantly in the activating blood and supplementing qi group, Perindopril group and Tongxinluo group compared with the model group (P<0.01). Conclusion The supplementing qi and activating blood circulation herbs can improve regional myocardial blood supply by decreasing the number of pericytes and promoting regeneration of capillary.

Objective To investigate insulin resistance and the effect of physical training on it in the pa- tients with chronic heart failure (CHF). Methods One hundred and twenty NYHAⅡ-ⅢCHF patients were ran- domly divided into a training group( n = 65 ) and a routine therapy group (n = 55 ). Another 35 healthy subjects were recruited as control group. All the patients were treated with routine anti-CHF drugs, and the training group patients had received physical training twice a day in addition. The HOMA-IR, insulin sensitivity index (ISI) , left ventricu- lar ejection fraction (LEVF), left ventricular fractional shortening( LVFS), 6-minute walking distance, heart rate and mean blood pressure were compared between the training and routine therapy groups before and after physical ex- ercise in both groups, and a comparison was made between the patients and the controls before the intervention with regard to HOMA-IR and ISI. Results Comparing with control group, ISI was reduced while the HOMA-IR in- creased (P

OBJECTIVETo study the effects of the extensive resection of the tumor-loading segment and artificial humerus head replacement combined with perioperative rehabilitation for the treatment of stage II to III giant cell tumor of bone in the proximal humerus.

METHODSFrom March 2007 to March 2010, 7 patients with stage II to III giant cell tumor of bone in the proximal humerus were treated. Among the patients, 3 patients were male and 4 patients were female with a mean age of 34.6 years (ranged, 18 to 49 years). The mean course of disease was 19 months (ranged, 6 to 35 months). All the patients were confirmed to suffer stage II to III giant cell tumor of bone in the proximal humerus by pathology and X-ray examinations. Clinical manifestations of the patients included persistence aggravated pain of the shoulder, swelling in the proximate arm with obviously tenderness, activity limited of the joint. All the patients were treated with extensive resection of the tumor-loading segment and artificial humerus head replacement combined with perioperative rehabilitation. CMS and OSIS score system were used to evaluate shoulder function and shoulder stability.

RESULTSAll the patients were followed up, and the duration ranged from 14 to 35 months, with an average of 17 months. There were no serious complications or recurrence in all cases. One year after the surgery CMS and OSIS score system were 70.7 scores (ranged,63 to 82 scores) and 25.1 scores (ranged, 18 to 29 scores) respectively. According to evaluation for shoulder function, 2 patients got an excellent result and 5 good. According to evaluation of shoulder stability, 1 patient got an excellent result and 6 good.

CONCLUSIONExtensive resection of the tumor-loading segment and artificial humerus head replacement combined with perioperative rehabilitation for the treatment of stage II to III giant cell tumor of bone in the proximal humerus would not only preserve the upper extremity but also preserve the function of upper extremity.

Adolescent ; Adult ; Bone Neoplasms ; pathology ; rehabilitation ; surgery ; Female ; Giant Cell Tumor of Bone ; pathology ; rehabilitation ; surgery ; Humans ; Humerus ; surgery ; Limb Salvage ; Male ; Middle Aged ; Neoplasm Staging

Objective: To delineate the variants spectrum of phenytalanine hydroxylase (PAH) gene among 78 unrelated patients with phenylketonuria (PKU) from Jiangxi province. Methods: The 13 exons and flanking intronic regions of the PAH gene were subjected to PCR amplification and sequencing. Results: A total of 143 variants were detected among the 156 alleles, which included 54 types of variants, which yielded a detection rate of 91.7%. Common variants have included R243Q (26/143, 18.2%), R408Q (10/143, 7.0%), EX6-96A>G (8/143, 5.6%), IVS4-1G>A (7/143, 4.9%), R241C (7/143, 4.9%) and V399V (7/143, 4.9%). In addition, 6 novel variants were detected, which included IVS4-3T>G, Q172H, C284Y, V291L, V329del, and L430R. The variants consisted of missense, splicing, nonsense and deletion variants, which have mainly located in exons 7 (45, 31.5%), 12 (17, 11.9%), 11 (16, 11.2%) and 6 (14, 9.8%). Conclusion Variants of the PAH gene identified in Jiangxi province mainly involve exons 7, 12, 11 and 6, with the most common variants being R243Q and R408Q. Six novel variants were identified.

The expression of the vgb gene in vivo could improve the fermentation density and then contribute the extracellular secretion of the product of bxn gene. Constructed the recombination plasmid pPIC9K-vgbbxn and transformed into Pichia pastoris GS115. The results of PCR and SDS-PAGE indicate that the vgb gene and bxn gene had integrated into the genome of Pichia pastoris GS115 and expressed in efficient level. Also, the protein activity of their products had been verified respectively. Shake flask fermentation experiments showed that the presence of VHb in yeast Pichia pastoris efficiently enhanced cell growth and secretive expression of bxn gene under hypoxic habitats.
Aminohydrolases ; genetics ; metabolism ; Bacterial Proteins ; genetics ; physiology ; Electrophoresis, Polyacrylamide Gel ; Pichia ; genetics ; Plasmids ; Polymerase Chain Reaction ; Recombinant Proteins ; biosynthesis ; Truncated Hemoglobins ; genetics ; physiology

Professor LUO Cai-gui's experience of acupuncture at acupoint "Baliao" with twisting manipulation for treatment of low back pain is introduced. This method has significant efficacy on improving low back pain and numbness of lower extremities, which is characterized with short-time manipulation, quick de-qi and long effective time. The acupuncture methods, manipulations, precautions, etc. are elaborated in details. A typical case is added.
Acupuncture Points ; Acupuncture Therapy ; instrumentation ; methods ; Humans ; Low Back Pain ; therapy ; Male ; Middle Aged

BACKGROUNDAppropriate expression and regulation of the transcriptome, which mainly comprise of mRNAs and lncRNAs, are important for all biological and cellular processes including the physiological activities of bone microvascular endothelial cells (BMECs). Through an intricate intracellular signaling systems, the transcriptome regulates the pharmacological response of the cells. Although studies have elucidated the impact of glucocorticoids (GCs) cell-specific gene expression signatures, it remains necessary to comprehensively characterize the impact of lncRNAs to transcriptional changes.

METHODSBMECs were divided into two groups. One was treated with GCs and the other left untreated as a paired control. Differential expression was analyzed with GeneSpring software V12.0 (Agilent, Santa Clara, CA, USA) and hierarchical clustering was conducted using Cluster 3.0 software. The Gene Ontology (GO) analysis was performed with Molecular Annotation System provided by CapitalBio Corporation.

RESULTSOur results highlight the involvement of genes implicated in development, differentiation and apoptosis following GC stimulation. Elucidation of differential gene expression emphasizes the importance of regulatory gene networks induced by GCs. We identified 73 up-regulated and 166 down-regulated long noncoding RNAs, the expression of 107 of which significantly correlated with 172 mRNAs induced by hydrocortisone.

CONCLUSIONSTranscriptome analysis of BMECs from human samples was performed to identify specific gene networks induced by GCs. Our results identified complex RNA crosstalk underlying the pathogenesis of steroid-induced necrosis of femoral head.

Cells, Cultured ; Endothelial Cells ; drug effects ; metabolism ; Femur Head ; cytology ; Gene Expression Profiling ; Glucocorticoids ; pharmacology ; Humans ; Oligonucleotide Array Sequence Analysis ; Osteonecrosis ; genetics ; RNA, Messenger ; genetics ; RNA, Untranslated ; genetics ; Transcriptome ; drug effects ; genetics

OBJECTIVETo analyze the clinical and epidemiological characteristics of Dengue fever (DF) during the Dengue-1 epidemic in Guangzhou.

METHODSClinical and epidemiological data of 1032 patients with DF from May 2002 to November 2003 were retrospectively analyzed. Dengue virus were isolated by cell culture and typed by reverse transcriptase polymerase chain reaction (RT-PCR).

RESULTSAge of the patients ranged from 55 days to 91 years old (average 34.7 +/- 13.2 years) with sex ratio 1.03:1. Incubation period ranged from 2 to 12 days with mean periods of 5.3 +/- 2.4 days. Most (45.0%) cases appeared in September and the epidemic last from July to November. Dengue outbreak had involved 675 cases in 26 common places. The common manifestations were seen as fever (100%), headache (90.9%), myalgia (68.4%), bone soreness (48.8%), fatigue (79.3%), skin rash (60.1%), positive tourniquet test (45.3%), leukopenia (63.3%) and thrombocytopenia (60.8%), respectively. Dengue virus was isolated from serum of 19 out of 54 patients' and identified as Dengue virus type 1. DNA sequence analyzes on rates of nucleotide homology were 97%, 97% and 98% compared with those of Dengue virus type 1 strain of DF outbreak in Cambodia, in 1997 and 1999 in China.

CONCLUSIONThe epidemic of DF in Guangzhou in 2002/2003 was caused by Dengue virus type-1 with most patients showing classic type of the disease. Date suggested that change can happen from non-endemic to hypoendemic regions in Guangdong province.

Adolescent ; Adult ; Aged ; Aged, 80 and over ; Base Sequence ; Child ; Child, Preschool ; China ; epidemiology ; Dengue ; epidemiology ; Dengue Virus ; classification ; genetics ; isolation & purification ; Female ; Humans ; Incidence ; Infant ; Male ; Middle Aged ; Molecular Sequence Data