Animals ; Concanavalin A ; adverse effects ; Immunoglobulin G ; blood ; Interleukin-18 ; blood ; immunology ; pharmacology ; Liver Cirrhosis ; blood ; chemically induced ; prevention & control ; Mice ; Mice, Inbred BALB C

Objective To compare the differences of clinical characteristics between genotype B and C chronic hepatitis B(CHB)patients and to summarize clinical factors related to genotype C hepa- titis B virus(HBV)infection.Methods Seventy eight CHB patients who were diagnosed with genotype B or C infection by liver puncture biopsy and genotyping were enrolled.Their serum HBV DNA levels were detected.Severe hepatitis,liver cirrhosis,hepatocellular carcinoma and HBeAg positive rate were analyzed to determine the pathologic inflammation and fibrosis degree of liver tissue.Chi square test and Logistic multiple regression analysis were employed for the statistical analysis.Results The serum albumin and pre-protein were lower in genotype C CHB patients than that in genotype B.The alanine aminotrans- ferase,total bilirubin and prothrombin time were higher in genotype C CHB patients than that in genotype B.The rates of genotype C patients increased significantly with the grade of liver necroin- flammation progressing from GO to G4(1.8%,11.1%,20.4%,33.3%,33.3%) and the stage of liver fibrosis progressing from SO to S4(5.6%,5.6%,14.8%,33.3%,40.7%),but the rates of genotype B patients did not change significantly with the grade of liver necroinflammation(16.7%, 25.0%,25.0%,20.8%,12.5%)and stage of liver fibrosis progressing(16.7%,29.2%%,20.8%, 16.7%,16.7%).There was statistical significance in grades of liver necroinflammation(X~2= 11.49,P=0.022)and stages of liver fibrosis(X~2=13.56,P=0.006)between genotype B and gen- otype C patients.The rates of genotype C CHB patients were higher than,similar with and lower than the rates of genotype B patients of HBV DNA level above 1.0?10~6 copy/mL,between 5.0?10~2-1.0?10~6 copy/mL and under 5.0?10~2 copy/mL,respectively(51.8% vs 12.5%,35.2% vs 45.8% and 13.0% vs 41.7%).There was statistical significance of HBV loads between genotype B and genotype C patients(X~2=13.25,P=0.001).HBeAg positive rate in genotype C patients was significantly higher than that in genotype B patients(61.1% vs 25.0%,X~2=8.67,P=0.003).The rates of decompensated cirrhosis,compensated cirrhosis and no-cirrhosis in genotype C patients were higher than,similiar with and lower than the rates in genotype B patients,respectively(40.7% vs 4.2%,22.2% vs 20.8% and 37.0% vs 75.0%).There was statistical significance of the rate of cirrhosis between genotype B and genotype C patients (X~2=12.47,P=0.002).Conclusions The degree of liver necroinflammation and fibrosis,the HBeAg positive rate and the incidence of cirrhosis are all related with genotype C HBV infection.

OBJECTIVETo establish a HPLC-based method for simultaneous determination of 2 classes of compounds (flavonoids and chromones) and 6 their effective components,(including prin-O-glucosylcimifugin, cimifugin, 4'-O-beta-D-glucosyl- 5-O-methylvisamminol, quercetin, sec-o-glucosylhamaudol and formononetin), in Yupingfeng Decoction.

METHODSHPLC-based separation of the agents was performed on Agilent Extend-C(18) column (4.6 mm x 250 mm, 5 microm) at 25 degrees with the mobile phase of MeOH-1% acetic acid water solution (gradient elution), flow rate of 0.8 ml/min and detection wavelength of 254 nm.

RESULTS AND CONCLUSIONHPLC allowed simultaneous quantitative determination of the 6 components in Yupingfeng Decoction, and they showed good linear relationships when their sample amount ranged 90-1810 ng, 97-1940 ng, 190-1906 ng, 105-3144 ng, 88-2625 ng and 109-3279 ng, respectively, with correlation coefficients all beyond 0.9999 and average recovery rates of 98.2%, 99.1%, 97.3%, 97.8%, 98.8% and 99.2%, respectively. This simple and convenient method accommodated a broad linear range with high sensitivity and precise and reproducible results.

Chromatography, High Pressure Liquid ; methods ; Chromones ; analysis ; Drugs, Chinese Herbal ; chemistry ; Flavonoids ; analysis ; Isoflavones ; analysis ; Quercetin ; analysis ; Reproducibility of Results

The 1H-NMR fingerprints of three different species tibetan medicine sea buckthorn were established by 1H-HMR metabolomics to find out different motablism which could provide a new method for the quality evaluation of sea buckthorn. The obtained free induction decay (FID) signal will be imported into MestReNova software and into divide segments. The data will be normalized and processed by principal component analysis and.partial least squares discriminant analysis to perform pattern recognition. The results showed that 25 metabolites belonging to different chemical types were detected from sea buckthorn,including flavonoids, triterpenoids, amino acids, carbohydrates, fatty acids, etc. PCA and PLS-DA analysis showed three different varietiest of sea buckthorn that can be clearly separated by the content of L-quebrachitol, malic acid and some unidentified sugars, which can be used as the differences metabolites of three species of sea buckthorn. 1H-NMR-based metabonomies method had a holistic characteristic with sample preparation and handling. The results of this study can offer an important reference for the species identification and quality control of sea buckthorn.
Hippophae ; metabolism ; Magnetic Resonance Spectroscopy ; methods ; Medicine, Tibetan Traditional ; Metabolomics

For in vitro studies, both CD34+ selected cell and mononuclear cell (MNC) can be used to expand hematopoietic stem/progenitor cells. To investigate the expansion characteristics of mononuclear cells (MNC) and CD34+ selected cells the two cell fractions were cultured in the medium containing cytokine cocktails of SCF + IL-3 + IL-6 + FL + Tpo. It was found that the CD34+ selected cells had presented a high proliferation potential. The expansion of CD34+ selected cells could be maintained for 8 weeks while that of MNCs declined after 4 weeks. During the culture period, the maximum expansion of total cells in CD34+ selected cell culture achieved 31,270.9 +/- 8640.5 times, while that of MNC reached 50.9 +/- 8.2 times only. In the culture of MNCs, the colony density and the proportion of CD34+ cells increased from day 0 to day 7. However, in the culture of CD34+ selected cells, both the colony density and the proportion of CD34+ cells declined continuously during the whole culture period. During the ex vivo culture of CD34+ selected cells, the maximum expansion of CFU-GM and CD34+ cells achieved 185.7 +/- 14.1 fold and 191.7 +/- 188.8 fold, respectively. They are much higher than that of MNC, which were 12.4 +/- 3.2 fold and 50.6 +/- 33.2 fold only. While the BFU-E of both cell fractions only expanded by few times, which were 7.2 +/- 5.2 and 10.1 +/- 3.4 times, respectively. The results showed that the CD34+ selected cells culture could obtain more CFU-GM cells and CD34+ cells during the whole culture period.
Antigens, CD34 ; analysis ; Cell Count ; Cell Separation ; Cells, Cultured ; Fetal Blood ; cytology ; Hematopoietic Stem Cells ; physiology ; Leukocytes, Mononuclear ; cytology

OBJECTIVETo observe the application effect of minimally invasive decompression, bone graft implantation and metal trabecular bone reconstruction system for early stage osteonecrosis of femoral head and discuss the treatment of hip-salvage operation in early stage osteonecrosis of femoral head;

METHODSFrom January 2010 to June 2011, 50 patients (62 hips) Which were osteonecrosis of femoral head of early stake,were treated with minimally invasive decompression, bone graft implantation and metal trabecular bone reconstruction system, including 31 males (40 hips), 19 females (22 hip) with an average age of 36.2 years old ranging from 22 to 54 years old. The course of disease was from 6 to 15 months (averaged 10.5 months). Among them, 19 cases (23 hips) were steroid-induced, 25 cases (33 hips) were alcohol-induced, 6 cases (6 hips) were idiopathic; According to ARCO stage, 28 hips were at stage I, 34 hips were at stage II. All of them were diagnosed as femoral head necrosis by imaging examination before operation. Then each patient was followed to assess by Harris hip score, curative effect, and conduct the femoral head survival analysis during the postoperation.

RESULTSAll patients had finished operation, the operation time was between 30 and 85 min, intraoperative blood loss was 50 to 220 ml, and 47 cases (58 hips) were follow-up from 24 to 46 months with an average of 34.05 months. As compared with preoperative, the Harris hip score at the last follow-up was improved, the difference was statistically significant (P<0.01). The Harris hip score, curative effect and survival time of femoral head in ARCO stage I was superior to these in ARCO Stage II, the difference was statistically significant (P< 0.05).

CONCLUSIONEffect of minimally invasive decompression,bone graft implantation combine with the metal trabecular bone reconstruction system for early stage osteonecrosis of femoral head was good,it could significantly improve the Harris hip score, increase the femoral head survival time, delay the hip replacement, and performance better in ARCO stage I.

Adult ; Bone Transplantation ; Decompression, Surgical ; Female ; Femur Head ; injuries ; pathology ; surgery ; Femur Head Necrosis ; surgery ; Follow-Up Studies ; Humans ; Male ; Middle Aged ; Prostheses and Implants ; Reconstructive Surgical Procedures ; Young Adult

OBJECTIVETo observe the curative effect of porous tantalum rod and Gugutou Huaisiyu Capsule (GHC) for steroid-induced osteonecrosis of femoral head (SONFH).

METHODSA total 60 hips of 50 SONFH patients were randomly assigned to the treatment group and the control group according to grouping time, 25 in each group (30 hips). Patients in the control group were implanted with porous tantalum rod, while those in the treatment group additionally took GHC (5 pills each time, three time per day for 2 successive months; and then twice per day for 4 successive months). Then all patients were followed-up to observe Harris hip score. The curative effect and the femoral head survival time were assessed.

RESULTSA total of 49 patients (59 hips) were followed-up. The Harris hip score of the two groups at the final follow-up was significantly improved after treatment, with statistical difference when compared with before treatment (P < 0.01). Besides, it was higher in the treatment group than in the control group. The curative effect and the survival time were superior in the treatment group, with statistical difference when compared with the control group (P < 0.05).

CONCLUSIONSPorous tantalum rod combined GHC got better effect in treating SONFH. It could significantly improve the function of affected hips and prolong the survival time of femoral head.

Capsules ; Drugs, Chinese Herbal ; administration & dosage ; pharmacology ; therapeutic use ; Femur Head Necrosis ; drug therapy ; Humans ; Prostheses and Implants ; Steroids ; adverse effects ; Tantalum

Objective To observe the influence of long-term treatment with inhaled corticosteroid on biochemical bone indexes and bone mineral density (BMD) in children with asthma. Methods The design was a randomized, paralleled group study with 3 low dose regiments of 100, 200,300 micrograms of budesonide per day in 45 children with asthma aged 5-8 years old for 12 months. Before inhaled corticosteroid therapy and 6th,12th month,clinical effects were observed and lung function(FEV1) was measured; concentration of serum osteocalcin(OST),insulin-like growth factor-1(IGF-1),bony alkaline phosphatase (BALP) and urinary deoxypyridinolin: creatinine (DPD/Cr) were measured; BMD was examined by dual energy X-ray absorptiometry. Results Clinical evaluation was improved and there was significant increase in FEV1 of asthmatic children. The amount of serum OST was slightly higher,yet no significant compared with that of normal control group. There was significant increase of serum BALP in asthmatic children after treatment; there was significant increase in serum IGF-1 of patients group after treatment compared with in normal children at the same age group; there was significant decrease in urinary DPD/Cr after treatment.There was no significant decrease in BMD before and after treatment at the hip (neck of femur , trochanter of femur ,Ward′s triangle),the lumber area of the spine (L2-4) and forearm (ultradis, distal). Conclusion Long-term treatment with low does corticosteroid dose not restrictedly affect bone metabolism and BMD in children with asthma.

Stirred culture offers a number of advantages over static systems as it maintains a stable, homogeneous culture environment and is easy to scale-up. This paper focused on the development and application of stirred tank bioreactor to culture hematopoietic cells. Preliminary study of stirred culture of hematopoietic cells was carried out in cord blood mononuclear cells culture in spinner flask. The results showed that the amplification rates of total cell, CFU-GM and BFU-E, with the exception of CFU-Mk, were greater in spinner flask than T-flask. The number of total cells increased 20 fold after 14 days incubation in spinner flask. The amplification rates of CFU-GM, CFU-Mk and BFU-E reached maximum at 10th day, 10th day and 7th day respectively, and the maximal amplification rates were 9.2-fold, 5.5-fold and 2.4-fold respectively, whereas the rate of CD34+ cells in spinner flask was (6.7 +/- 4.0)-fold at day 10. These results indicated that the stirred culture system is better than the static culture systems for hematopoietic cell proliferation. The biocompatibility of cord blood MNC to different types of materials used in bioreactors was also tested. The results showed that glass, stainless steel 316L and polytetraflouroethylene (PTFE) supported the growth of hematopoietic cells well. A higher cell density was reached in stirred bioreactors with controlled pH and DO than static culture. These findings suggested that the controlled large-scale culture could be used to overcome the clinical shortage of hematopoietic cells.
Antigens, CD34 ; metabolism ; Bioreactors ; Cell Culture Techniques ; instrumentation ; methods ; Erythroid Precursor Cells ; cytology ; Fetal Blood ; cytology ; Granulocyte-Macrophage Progenitor Cells ; cytology ; Humans ; Polytetrafluoroethylene ; Stainless Steel

Clinical evidence of hematopoietic restoration with umbilical cord blood (UCB) grafts indicates the UCB can be a useful source of hematopoietic stem cells for routine bone marrow reconstitution. Considering (10 +/- 5) x 10(8) nucleared cells per cord blood unit, there is a potential limitation for the use of cord blood in adults, which, however, can be overcome by ex vivo expansion of cells. A prerequisite for expansion is the significantly higher recovery of MNC, CD34+ cells and colony-forming cells (CFC) by thawing cryopreserved MNC. Cooling rate always acts as a critical factor that can affect the recovery of cells. Although the rate of - 1 degrees C/min is adopted in most of the cryopreservations, no data has been reported about the detailed effects of different cooling rates. The aim of the study was to reveal the different effects of cooling rates on cryopreservation of hematopoietic stem cells from cord blood. UCB samples were collected, and cryopreserved as mononuclear cells (MNC) with different cooling rates of - 0.5 degrees C/min, - 1 degrees C/min, - 5 degrees C/min, and the recovery and viability of MNC and CD34+ cells, the clonogenic capacity and the ex vivo expansion potential of UCB progenitor cells were evaluated after thawing. With - 1 degrees C/min cooling rate, the recovery of MNC reached 93.3% +/- 1.8% , viability 95.0% +/- 3.9% , recovery of CD34+ cells 80.0% +/- 17.9% , and clonogenic recovery were 87.1% +/- 5.5%, 88.5% +/- 8.9%, 86.2% +/- 7.4% for BFU-E CFU-GM CFU-MK, respectively. After 14 days of liquid culture, no significant difference was detected in CFC expansion between fresh and cryopreserved MNC cells with - 1 degrees C/min cooling rate, but this was not the case with - 0.5 degreesC/min and - 5 degrees C/min. In conclusion, it was demonstrated that controlling the rate at - 1 degrees C/min is more suitable for cryopreservation of hematopoietic stem cells than - 0.5 degrees C/min and - 5 degrees C/min.
Cell Survival ; physiology ; Cells, Cultured ; Cryopreservation ; methods ; Erythroid Precursor Cells ; cytology ; Fetal Blood ; cytology ; Flow Cytometry ; Granulocyte-Macrophage Progenitor Cells ; cytology ; Hematopoietic Stem Cells ; cytology ; Humans