OBJECTIVETo develop the procedure for separating the ethanol-soluble and acidic components (ESAC) from Ganoderma lucidum, and to establish a method for quantifying ESAC in G. lucidum.

METHODThe ethanol extract of G. lucidum was extracted with a saturated NaHCO3 solution, acidified and re-extracted by chloroform to obtain ESAC. The quantitative analysis of ESAC was based on the characteristic color reaction between ESAC and H2SO4.

RESULTThe optimal conditions for separating ESAC on a 10 g scale are as follows: ratio of material and ethanol (mL), 1:15; immersing time, 24 h; volume of saturated NaHCO3 and chloroform, 1300 mL; extract 3 times. The condition for measuring ESAC is as follows: sample weight, 1 g; solution volume, 1.5 mL; immuersing time, 0.5 h; detecting reagent, 50% H2SO4 in ethanol; heating time in 100 degrees C water bathe, 3 min; measuring wavelength, 490 nm.

CONCLUSIONThe procedure for ESAC preparation is simple and well-designed, and the established method for ESAC can be used for the qualitative analysis of the G. lucidum related products.

Drugs, Chinese Herbal ; analysis ; isolation & purification ; Ethanol ; Reishi ; chemistry ; Technology, Pharmaceutical ; methods ; Triterpenes ; analysis ; isolation & purification

OBJECTIVETo observe the change of the salivary occult blood after periodontal mechanical therapy, and to assess the correlations between salivary occult blood and the level of volatile sulphur compounds (VSC) in oral cavity, periodontal clinical parameters, respectively.

METHODSFifty patients with gingivititis, mild or moderate periodontitis were included. The level of VSC were measured by Halimeter(®) and salivary occult blood was tested by Perioscreen(®) before periodontal examination. Then full mouth plaque index (PLI), probing depth (PD), bleeding index (BI) were charted. Attachment loss (AL) of the Ramfjörd teeth were recorded lastly. Intensive periodontal mechanical therapy was conducted including oral hygiene instruction, scaling and root planing (SRP). Four weeks after SRP, the same examinations were repeated.

RESULTSSalivary occult blood was significantly correlated with BI (r = 0.294) and PLI (r = 0.308) before periodontal therapy (P < 0.01), and also significantly correlated with VSC level (r = 0.386), PLI (r = 0.456), BI (r = 0.352), AL (r = 0.325) after therapy (P < 0.05). The improvement of VSC level [211.0 (111.0 - 389.5) × 10⁻⁹ vs 100.0 (46.3 - 165.3) × 10⁻⁹], the clinical periodontal parameters including PLI [(1.3 ± 1.0) vs (0.4 ± 0.6)], PD [(3.7 ± 1.5) mm vs (2.7 ± 0.9) mm], BI [(1.8 ± 1.2) vs (0.4 ± 0.7)] and AL [(1.0 ± 1.1) mm vs (0.1 ± 0.5) mm after the treatment was statistically significant (P < 0.001). However, standing on the viewpoint of salivary occult blood changes from positive before therapy to negative after therapy, only 80% (40/50) individuals were totally cured. VSC level in oral cavity and periodontal clinical parameters significantly decreased (P < 0.001) following the trends from strong positive, weak positive, to negative results of salivary occult blood test.

CONCLUSIONSSalivary occult blood was correlated with VSC level in oral cavity of periodontal treated patients. It may be an objective parameter to evaluate the gingival inflammation and the efficacy of the periodontal therapy at individual level.

Dental Plaque Index ; Gingivitis ; blood ; therapy ; Humans ; Mouth ; chemistry ; Occult Blood ; Periodontitis ; blood ; therapy ; Saliva ; chemistry ; Sulfur Compounds ; chemistry ; Volatile Organic Compounds ; chemistry

OBJECTIVETo observe the effects of fenvalerate (Fen) on ovarian calcium homeostasis.

METHODShGLCs were obtained from pre-ovulatory follicles in an in vitro fertilization program, and were cultured for 72 hours. Changes in cellular [Ca(2+)]i induced by Fen in hGLCs were detected with laser scanning confocal microscopy (LSCM) by using the fluorescent Ca(2+) indicator fluo-3/AM. SD female rats were divided into four groups (control, 1/15LD(50), 1/50 LD(50) and 1/250 LD(50)) in experiment. The activity of ovarian Ca(2+)-ATPase and phosphorylase A (P-a) and the contents of calmodulin (CaM) were assessed after a 30-day Fen exposure. In addition, serum estradiol-17 beta (E(2)) and progesterone (P(0)) concentration were measured by radioimmunoassay, which the sampling rats were ensured at diestrus stage before killed according to vaginal smear.

RESULTS20.0 and 2.0 micromol/L Fen induced the increased of [Ca(2+)]i in hGLC. This [Ca(2+)]i increase mostly resulted from Ca(2+) influx in the studied concentration. Fen had shown the inhibition effects on activity of Ca(2+)-ATPase in 1/250 LD(50) group (P < 0.001) while the activity of phosphorylase A (P-a) in treated groups had significantly enhanced than those of in control. The contents of CaM in ovaries were found to be increased in treated groups. E(2) in 1/250 LD(50) group were higher while P(0) in 1/15 LD(50) group were significantly lower (P < 0.05).

CONCLUSIONExposure to Fen interferes the serum steroid hormone concentrations partly through calcium signal pathway.

Adenosine Triphosphatases ; metabolism ; Animals ; Calcium ; metabolism ; Cells, Cultured ; Female ; Gonadal Steroid Hormones ; blood ; Granulosa Cells ; drug effects ; metabolism ; Homeostasis ; drug effects ; Humans ; Insecticides ; toxicity ; Nitriles ; Ovary ; cytology ; drug effects ; metabolism ; Pyrethrins ; toxicity ; Rats ; Rats, Sprague-Dawley

OBJECTIVETo explore the recent clinical curative effect of Tantalum rod in treating the early avascular necrosis.

METHODSFrom January 2008 to November 2008, the 25 patients (39 hips) with early avascular necrosis accepted tantalum rod placement and included 9 males (11 hips) and 16 females (28 hips) with an average age of 37 years old ranging from 18 to 74 years old. Four patients (6 hips) caused by Alcoholic, 6 patients (8 hips) by hormone, 2 cases (2 hips) by traumatic, 13 cases (23 hips) by idiopathic. Steinberg preoperative stage involved 7 hips in period I, 24 hips in period II, 8 hips in period III. Curative effect analysis included preoperative and postoperative Harris score, radiographic changes and hip replacement for follow-up to accept the end of the femoral head survival rate.

RESULTSAll patients were followed up for 6 to 47 months (averaged 37.4 months). All 12 hips imaging appeard progress,including tantalum rod exit in 1 hip, hip hemiarthroplasty collapse in 3 hips, the area increased to avascular necrosis in 8 hips. Six hips accepted total hip replacement, including imaging progress in 5 hips (41.7%, 5/12), no imaging progress in 1 hip (3.7%,1/27). All hips' Kaplan-Meier survival curves showed 6-month survival rate was (97.4 +/- 2.5)% after tantalum stick insertion, 1-year survival rate was (94.7 +/- 3.6), and 2-year survival rate was (88.6 +/- 5.4)%, 3-year survival rate was (72.5 +/- 11.2).

CONCLUSIONIt is effective for treatment of avascular necrosis of femoral head in Steinberg I and II by Tantalum rod, and it can effectively relieve femoral head replacement time.

Adolescent ; Adult ; Aged ; Female ; Femur Head Necrosis ; mortality ; surgery ; Humans ; Kaplan-Meier Estimate ; Male ; Middle Aged ; Prostheses and Implants ; Retrospective Studies ; Survival Rate ; Tantalum ; Time Factors ; Treatment Outcome ; Young Adult

OBJECTIVETo compare clinical effects of core decompression with stem cell transplantation and tantalum rod implanting in treating stage II non-traumatic osteonecrosis of femoral head.

METHODSFrom March 2012 to September 2012, 45 patients(55 hips)with stage ARCO II non-traumatic osteonecrosis of femoral head were treated and divided into core decompression with stem cell transplantation group(group A) and tantalum rod implanting group(group B) according to number table. In group A, there were 23 cases(28 hips) , including 12 males and 11 females aged from 23 to 51 years old with an average of (36.87±9.52) years, the courses of disease ranged from 2 to 28 months with an average of (17.13±7.74) months, preoperative Harris score was for 35 to 70 with an average of(54.74±11.81), treated with core decompression with stem cell transplantation. In group B, there were 22 cases(27 hips), including 11 males and 11 females aged from 26 to 46 years old with an average of (35.59±7.39) years, the courses of disease ranged from 3 to 26 months with an average of(16.00±7.46) months, preoperative Harris score was for 35 to 76 with an average of (57.18±12.95), treated with core tantalum rod implanting. Operative time, blood loss, hospital stays, hospitalization expenses were observed and compared after treatment between two groups, the clinical effects were evaluated according to Harris criteria.

RESULTSAll patients were followed up from 6 to 12 months with an average of 10.8 months. There were significant difference in hospitalization expenses between two groups(<0.05), while there was no significant statistical difference in blood loss and hospital stay (>0.05). At the final following-up, Harris score in group A was(83.04±8.97), 6 cases obtained excellent results, 14 good, 2 good and 1 poor;while Harris score in group A was(84.41±9.94), and 9 cases obtained excellent results, 9 good, 3 good and 1 poor; there was no statistical meaning differences between two groups(>0.05).

CONCLUSIONSCore decompression with stem cell transplantation and tantalum rod implanting could both improve function of hip joint, while core decompression with stem cell transplantation had advantages of shorter operation time, less cost, and higher potency ratio. It is suitable for stage ARCO II non-traumatic femoral head necrosis.

This study was aimed to explore the effects of peptidoglycan (PGN) on proliferation and cell cycle of human bone marrow mesenchymal stem cells (MSCs). MSCs were isolated from human bone marrow by density gradient centrifugation. The purity of MSCs with the spindle fibroblastic morphology was identified by microphotography and the phenotypes were detected by flow cytometry (FCM). MSCs incubated with different doses of PGN (1, 10, 20 μg/ml) were used as test groups, and those incubated without PGN were regarded as control group. The isolated and cultured MSCs were inoculated into 96-well plates according to a certain concentration. Cell cycle was measured by flow cytometry after incubated with PGN for 72 hours. The results showed that the cell proliferation index was significantly increased in dose and time dependent manners after MSCs was incubated with PGN. Its effects on the proliferation of MSCs were highest in 10 μg/ml group. Compared with the control group, PGN could significantly decrease proportion of MSCs in G₀/G₁ phase and increase them in S and G₂/M phases (p < 0.05). It is concluded that PGN can promote more MSCs to enter the DNA synthesis phase and proliferate many much MSCs in dose and time dependent manners.
Bone Marrow Cells ; cytology ; Cell Cycle ; drug effects ; Cell Proliferation ; drug effects ; Cells, Cultured ; Flow Cytometry ; Humans ; Mesenchymal Stromal Cells ; cytology ; Peptidoglycan ; pharmacology ; Toll-Like Receptor 2

OBJECTIVETo observe the effects of low-intensity pulsed ultrasound (LIPUS) on repairing extracellular matrix in rabbit knee osteoarthritis and analyze its mechanism.

METHODSSixty adult female rabbits with an average weight of (2.0 ± 0.2) kg, were divided randomly into two groups (experimental group and control group, 30 rabbits in each group). All rabbits were replicated in right knees by Hulth method for knee osteoarthritis model. Two weeks after operation, the rabbits in experimental group were treated with LIPUS, and the ultrasonic frequency was (800 ± 5%)KHz and the maximum intensities of spatially averaged and time averaged (SATA) was (50 ± 10%) mw/cm2, for 1 time a day and every time 20 min, while the rabbits in control group were treated with sham LIPUS,the same operation with experimental group but without energy output. At the 2, 4, 8 weeks after treatment, 10 rabbits in each group were randomly killed for each time. The general changes of cartilage and its histopathological changes by HE staining were observed; the expression of collagen type II, proteoglycan, MMP-3, 7, 13 in cartilage were analyzed by immunohistochemical and RT-PCR technique; and the expression of NO in cartilage was analyzed by nitrate reduction method.

RESULTSOn the same observed time point, the damage degree of cartilage in experimental group was slighter than that of control group (P < 0.01), the expression of MMP-3, 7, 13 and NO in cartilage in experimental group was lower than that of control group (P < 0.01) while collagen type II and proteoglycan was higher than that of control group (P < 0.01).

CONCLUSIONLow-intensity pulsed ultrasound can repair the damaged cartilage by reducing the expression of MMP-3, 7, 13, inhibiting the secretion of NO and promoting the synthesis of collagen type II and proteoglycan in cartilage.

Animals ; Cartilage, Articular ; pathology ; Collagen Type II ; biosynthesis ; Extracellular Matrix ; metabolism ; Female ; Matrix Metalloproteinases ; analysis ; Nitric Oxide ; biosynthesis ; Osteoarthritis, Knee ; metabolism ; therapy ; Rabbits ; Ultrasonic Therapy ; methods

OBJECTIVETo study the effect of terephthalic acid (TPA) on lipid metabolism in Sprague-Dawley (SD) rats.

METHODSFive groups of SD rats that ingested 0%, 0.04%, 0.2%, 1%, and 5% TPA, respectively, were included in a 90-day subchronic feeding study. Effects of TPA on levels of serum protein, total cholesterol (TC), triglycerides (TG), high-density lipoprotein cholesterol (HDL), total antioxidative capability (T-AOC), superoxide dismutase (SOD) and malondialdehyde (MDA) were observed. Urine samples were collected and analyzed for concentration of ion.

RESULTSTPA decreased the level of serum T-AOC in a dose dependent manner. The contents of serum and bladder MDA significantly decreased in 1% and 5% TPA ingestion groups. Serum CuZn superoxide dismutase (CuZnSOD) lowered in groups of 0.2%, 1%, and 5% TPA. TPA subchronic feeding had no significant influences on serum TC, LDL or HDL, but increased serum TG, TP and ALB after administration of 0.04% and/or 0.2% TPA. Concentrations of urinary Ca2+, Mg2+, Na+, and K+ were elevated in 1% and 5% TPA groups.

CONCLUSIONAntioxidative potential decreased after TPA exposure. MDA increase in serum and bladder tissues was one of the most important reactions in rats which could protect themselves against TPA impairment. The decrease of serum CuZnSOD was related to the excretion of Zn2+.

Animals ; Antioxidants ; analysis ; Blood Proteins ; analysis ; Cholesterol ; blood ; Female ; Ions ; urine ; Lipid Metabolism ; drug effects ; Lipoproteins ; blood ; Male ; Malondialdehyde ; blood ; Phthalic Acids ; toxicity ; Rats ; Rats, Sprague-Dawley ; Superoxides ; blood ; Triglycerides ; blood ; Weight Gain

OBJECTIVETo investigate the expression and relationship between HSP70 and caspase-3 in knee osteoarthritis.

METHODSForty adult SD rats were divided into experimental group and control group. Thirty rats in experimental group, anterior cruciate ligament transaction (ACLT) was cut off and partial meniscectomy of 1/3 inside incision were performed to reproduce knee osteoarthritis (KOA) model according to Hulth methos, and the other 10 rats was treated with nothing as control group. The rats were sent to the cage and free to move. At 1, 2 and 4 weeks later, the arthritis cartilage of femoral and tibial end were observed through immunohistochemistry staining and light microscope. Meanwhile, Mankin scale system was adopted for histomorphology evaluation.

RESULTSChanges of KOA such as hyperplastic synovium,erosion on the surface of cartilage and so on were found in experiment group, the expression of HSP70 was augmentation all the time, but the expression of caspase-3 was reduction 1 week later; no similar changes were found in control group. Mankin scale system showed that there were significant differences in the first week as compared with the second week and 4th week (both P < 0.01).

CONCLUSIONHeat shock protein inhibit the apoptosis of cartilage cells and protect the cartilage cells in knee osteoarthritis, the conservative treatment for clinical provide objective scientific basis.

Animals ; Apoptosis ; Cartilage ; pathology ; Caspase 3 ; physiology ; Chondrocytes ; pathology ; Female ; HSP70 Heat-Shock Proteins ; physiology ; Male ; Osteoarthritis, Knee ; pathology ; Rats ; Rats, Sprague-Dawley

OBJECTIVETo explore the risk factors for relapse after allogeneic hematopoietic stem cell transplantation (allo-HSCT) and the measures of prophylaxis and treatment.

METHODSWe summarized the clinical data of 82 patients with hematologic malignancies who were treated in our hospital from August 2003 to December 2008. Factors including age, sex, ABO blood group disparity of donor and recipient as well as the type of donor, status of disease, HLA-match, conditioning regimen, whether or not having developed acute GVHD and chronic GVHD, infusion number of CD34(+) cells, relationship between CMV infection and relapse post-transplantation were considered and analyzed.

RESULTSSingle factor analysis indicated that there were five independent risk factors related with the disease relapse (P < 0.05), including status of disease, time of diagnosis to transplantation, acute graft versus host disease (aGVHD), conditioning regimen, and chronic graft versus host disease (cGVHD). Simultaneously, the type of donor was a substantial factor (P < 0.01), determined by multi-factor Cox regression analysis. Cox regression analysis determined that disease status (OR = 2.58, 95%CI 1.26 - 5.01, P = 0.01), time from diagnosis to treatment (OR = 1.98, 95%CI 1.11 - 3.63, P = 0.025) and cGVHD (OR = 3.74, 95%CI 1.96 - 7.97, P < 0.001) were major factors for relapse of the patients who had undergone transplantation.

CONCLUSIONSRelapse remains the primary cause of failure after allo-HSCT. Status of disease, time from diagnosis to treatment and not cGVHD are the major risk factors. Effective prevention and treatment of relapse after engraftment can improve the efficacy of HSCT.

Adolescent ; Adult ; Child ; Female ; Follow-Up Studies ; Graft vs Host Disease ; etiology ; Hematologic Neoplasms ; therapy ; Hematopoietic Stem Cell Transplantation ; adverse effects ; Humans ; Infection ; etiology ; Male ; Middle Aged ; Recurrence ; Risk Factors ; Time Factors ; Transplantation Conditioning ; Transplantation, Homologous ; Young Adult