Simutaneous determination of ursolic acid and oleanolic acid in Ziziphora clinopodioides Lam. by in situ pretreatment-thin layer chromatography scanning / 国际药学研究杂志

Objective To establish a method for the simultaneous determination of ursolic acid(UA)and oleanolic acid(OA) in Ziziphora clinopodioides Lam.,and the quantitative determination the UA and OA contents in the different Z. clinopodioides plant samples collected with various parts of the plant at different times,from different regions of Xinjiang,China. Methods Dual wave?length scanning method was used for the quantification of UA and OA spots on a silica gel G plate in the TLC analysis. The samples loaded on the silica gel G plate were in situ treated with the 1%iodine solution in dichloromethane,and then the plate was developed using cyclohexane,cyclohexane-chloroform-ethyl acetate-formic acid(20:5:8:0.1)as the developing solvent. In the dual wavelength scanning,the measurement wavelength was 530 nm and the reference wavelength was 700 nm. Results The UA and OA spots in samples were well separated on the TLC plate and could be simultaneously quantified by the present method. The average contents of UA and OA in Z. clinopodioides plant samples from 18 different areas were(1.84 ± 0.41)and(2.82 ± 0.89)mg/g,respectively. The contents of UA and OA in the plant increased from late spring to early summer and then decreased thereafter. As to the different parts of the plant,the contents of UA and OA were highest in leaves and lowest in stems. Conclusion The method is simple,fast and accu?rate. The present results provided basic data for further evaluation of the quality of Z. clinopodioides resources.

Clinical significance of direct antiglobulin testing in anemia in patients with severe chronic hepatitis B / 中国感染控制杂志

Objective To evaluate clinical significance of direct antiglobulin testing(DAT)in anemia in patients with severe chronic hepatitis B(CHB).Methods Red blood cell(RBC)-related parameters detection and DAT were performed among 30 healthy persons,30 CHB patients,and 50 severe CHB patients,clinical factors related to posi-tive DAT were analyzed.Results RBC count,hemoglobin (Hb)concentration,and hematocrit(HCT)level in severe CHB patients were all lower than CHB patients and healthy group(P <0.05),RBC distribution width(RDW)in severe CHB patients were all higher than CHB patients and healthy group(P<0.05);the positive rate of DAT in patients with se-vere CHB,CHB,and healthy group were 62.82%,13.33% and 0 respectively.RBC count,Hb concentration,and HCT level in severe CHB patients with positive DAT were all lower than severe CHB patients with negative DAT (all P <0.05),while RDW was higher than the latter (P=0.001);after RBC was separated through capillary,positive intensity of DAT of aged RBCs was higher than young RBCs in severe CHB patients (P <0.001);among severe CHB patients, DAT-positive and-negative patients differed in gender,age,alanine aminotransferase,total bilirubin,complement C3, C-reactive protein,and complication of diabetes(all P≤0.05).Conclusion Anemia in severe CHB patients may be re-lated to immune hemolysis of aged RBCs induced by antibody adsorption.

Clinical significance of direct antiglobulin testing in anemia in patients with severe chronic hepatitis B / 中国感染控制杂志

Objective To evaluate clinical significance of direct antiglobulin testing(DAT)in anemia in patients with severe chronic hepatitis B(CHB).Methods Red blood cell(RBC)-related parameters detection and DAT were performed among 30 healthy persons,30 CHB patients,and 50 severe CHB patients,clinical factors related to posi-tive DAT were analyzed.Results RBC count,hemoglobin (Hb)concentration,and hematocrit(HCT)level in severe CHB patients were all lower than CHB patients and healthy group(P <0.05),RBC distribution width(RDW)in severe CHB patients were all higher than CHB patients and healthy group(P<0.05);the positive rate of DAT in patients with se-vere CHB,CHB,and healthy group were 62.82%,13.33% and 0 respectively.RBC count,Hb concentration,and HCT level in severe CHB patients with positive DAT were all lower than severe CHB patients with negative DAT (all P <0.05),while RDW was higher than the latter (P=0.001);after RBC was separated through capillary,positive intensity of DAT of aged RBCs was higher than young RBCs in severe CHB patients (P <0.001);among severe CHB patients, DAT-positive and-negative patients differed in gender,age,alanine aminotransferase,total bilirubin,complement C3, C-reactive protein,and complication of diabetes(all P≤0.05).Conclusion Anemia in severe CHB patients may be re-lated to immune hemolysis of aged RBCs induced by antibody adsorption.

Shorter survival rate in varus-aligned knees after total knee arthroplasty.

PURPOSE: One long-held tenet of total knee arthroplasty (TKA) is that post-operative neutral limb alignment promotes implant durability. Recently, the concept of generic safe zone (0° ± 3°) has been challenged. This meta-analysis aimed to evaluate whether neutral alignment was superior to malalignment in long-term survival of TKAs. METHODS: The MEDLINE, Embase, Cochrane Library, China National Knowledge Infrastructure, Wan Fang Chinese Periodical, Google and reference lists of all the included studies were searched. Of the 1512 studies initially identified, ten met the eligibility criteria, including eight case-control studies and two cohort trials. Relative risks of implant failure were compared between post-operative neutrally aligned and malaligned knees. RESULTS: Post-operative malalignment showed higher failure rate of knee implants compared with neutral alignment (95 % CI 1.00-1.88, P = 0.05). Failure rate in knees with varus alignment was significantly higher than with neutral alignment (95 % CI 1.07-2.55, P = 0.02). There was no significant difference in the likelihood of implant failure between knees with valgus and neutral alignment (95 % CI 0.78-2.41, n.s.). No significant difference of failure rate was noted between neutral alignment and malalignment for fixed-bearing prothesis (95 % CI 0.94-1.95, n.s.) or rotating-platform prothesis (95 % CI 0.75-2.73, n.s.). There was no significant difference of failure rate between knees with neutral alignment and malalignment for studies with a mean follow-up of more than 10 years (95 % CI 0.81-2.01, n.s.) or studies using long-leg weight-bearing radiographs (95 % CI 0.79-1.79, n.s.). CONCLUSIONS: Post-operative varus alignment results in shorter survival rate after TKA. Not only neutral limb alignment but also the valgus alignment promotes implant durability. Neutral or valgus alignment rather than varus alignment is essential to achieve long-term survival of TKAs and patient satisfaction. LEVEL OF EVIDENCE: III.

Extramedullary versus intramedullary tibial alignment technique in total knee arthroplasty: A meta-analysis of randomized controlled trials.

The aim of this study was to establish whether the use of an extramedullary or intramedullary tibial cutting guide leads to superior mechanical leg axis and implant positioning. A meta-analysis of six randomized controlled trials including 350 knees was performed. For the mechanical axis, frontal tibial component angle and tibial slope, there were no significant differences in the mean values or the number of outliers (±3°) between the extramedullary and intramedullary groups. A reduced tourniquet time was associated with the intramedullary guide. No significant difference in the complication rate was noted between the two groups. Neither extramedullary nor intramedullary tibial alignment was more accurate in facilitating the tibial cut. Use of an intramedullary guide results in a shorter tourniquet time and exhibits a similar complication rate as the extramedullary guide.

Extramedullary versus intramedullary tibial alignment technique in total knee arthroplasty: A meta-analysis of randomized controlled trials

The aim of this study was to establish whether the use of an extramedullary or intramedullary tibial cutting guide leads to superior mechanical leg axis and implant positioning. A meta-analysis of six randomized controlled trials including 350 knees was performed. For the mechanical axis, frontal tibial component angle and tibial slope, there were no significant differences in the mean values or the number of outliers (±3°) between the extramedullary and intramedullary groups. A reduced tourniquet time was associated with the intramedullary guide. No significant difference in the complication rate was noted between the two groups. Neither extramedullary nor intramedullary tibial alignment was more accurate in facilitating the tibial cut. Use of an intramedullary guide results in a shorter tourniquet time and exhibits a similar complication rate as the extramedullary guide.

Dose-dependent effects of nicotine on proliferation and differentiation of human bone marrow stromal cells and the antagonistic action of vitamin C.

A range of biological and molecular effects caused by nicotine are considered to effect bone metabolism. Vitamin C functions as a biological antioxidant. This study was to evaluate the in vitro effects of nicotine on human bone marrow stromal cells and whether Vitamin C supplementation show the antagonism action to high concentration nicotine. We used CCK-8, alkaline phosphatase (ALP) activity assay, Von Kossa staining, real-time polymerase chain reaction and Western Blot to evaluate the proliferation and osteogenic differentiation. The results indicated that the proliferation of BMSCs increased at the concentration of 50, 100 ng/ml, got inhibited at 1,000 ng/ml. When Vitamin C was added, the OD for proliferation increased. For ALP staining, we found that BMSCs treated with 50 and 100 ng/ml nicotine showed a higher activity compared with the control, and decreased at the 1,000 ng/ml. Bone morphogenetic protein-2 (BMP-2) expression and the calcium depositions decreased at 100 and 1,000 ng/ml nicotine, while the addition of Vitamin C reversed the down regulation. By real-time PCR, we detected that the mRNA expression of collagen type I (COL-I) and ALP were also increased in 50 and 100 ng/ml nicotine groups (P < 0.05), while reduced at 1,000 ng/ml (P < 0.05). When it came to osteocalcin (OCN), the changes were similar. Taken all together, it is found that nicotine has a two-phase effect on human BMSCs, showing that low level of nicotine could promote the proliferation and osteogenic differentiation while the high level display the opposite effect. Vitamin C could antagonize the inhibitory effect of higher concentration of nicotine partly.

Antitumor and anti-angiogenesis effects of thymoquinone on osteosarcoma through the NF-κB pathway.

Thymoquinone (TQ), the predominant bioactive constituent derived from the medicinal spice Nigella sativa (also known as black cumin), has been applied for medical purposes for more than 2,000 years. Recent studies reported that thymoquinone exhibited inhibitory effects on the cell proliferation of several cancer cell lines. This study was performed to investigate the antitumor and anti-angiogenic effects of thymoquinone on osteosarcoma in vitro and in vivo. Our results showed that thymoquinone induced a higher percentage of growth inhibition and apoptosis in the human osteosarcoma cell line SaOS-2 compared to that of control, and thymoquinone significantly blocked human umbilical vein endothelial cell (HUVEC) tube formation in a dose-dependent manner. To investigate the possible mechanisms involved in these events, we performed electrophoretic mobility shift assay (EMSA) and western blot analysis, and found that thymoquinone significantly downregulated NF-κB DNA-binding activity, XIAP, survivin and VEGF in SaOS-2 cells. Moreover, the expression of cleaved caspase-3 and Smac were upregulated in SaOS-2 cells after treatment with thymoquinone. In addition to these in vitro results, we also found that thymoquinone inhibits tumor angiogenesis and tumor growth through suppressing NF-κB and its regulated molecules. Collectively, our results demonstrate that thymoquinone effectively inhibits tumor growth and angiogenesis both in vitro and in vivo. Moreover, inhibition of NF-κB and downstream effector molecules is a possible underlying mechanism of the antitumor and anti-angiogenic activity of thymoquinone in osteosarcoma.

Effect of calcium citrate on bone integration in a rabbit femur defect model.

OBJECTIVE: To explore effect of calcium citrate on bone integration in a rabbit femur defect model, and to compare the bone formation with different sizes by radiological and histological study. METHODS: Twenty-four male Japanese white rabbits were randomly divided into three groups (Group A, B, C) in this study. Under anesthesia, defects of four sizes (1.2, 1.5, 2.0 and 2.5 mm) were created in each of the rabbits. Commercially pure calcium citrate powder was placed inside the medullary compartment of the femur (Experimental), while in the contralateral femur (Control) nothing was implanted. The defects were analyzed using radiography and histological analysis by using Imagepro-Plus 6.0 software after animal was sacrificed at 4th(Group A), 6th(Group B) and 8th(Group C) weeks postoperatively. Four samples were analyzed for each size of defect and each healing period. RESULTS: The histological and the radiologic evaluation were performed after sacrification of all rabbits on postoperative 4th and 6th weeks, It showed significant difference between the experimental group and the control group when these defects were less than or equal to 2.0 mm. No statistical difference was observed when these defects were larger than 2.0 mm at all healing periods except at the 4th week. CONCLUSIONS: Calcium citrate affects the early periods of bone defects healing mechanism in Japanese white rabbits positively, especially when the defect is not too large. We suggest further studies on calcium citrate to determine the effects of various dosages, administration ways and the experimental time on the bone defects.

Intramedullary nailing of clavicular midshaft fractures in adults using titanium elastic nail.

OBJECTIVE: Studies showed elastic stable intramedullary nailing (ESIN) of displaced midclavicular fractures has excellent outcomes, as well as high complication rates and specific problems. The aim was to discuss ESIN of midshaft clavicular fractures. METHODS: Totally 60 eligible patients (aged 18-63 years) were randomized to either ESIN group or non-operative group between January 2007 and May 2008. Clavicular shortening was measured after trauma and osseous consolidation. Radiographic union and complications were assessed. Function analysis including Constant shoulder scores and disabilities of the arm, shoulder and hand (DASH) scores were performed after a 15-month follow-up. RESULTS: ESIN led to a signifcantly shorter time to union, especially for simple fractures. In ESIN group, all patients got fracture union, of which 5 cases had medial skin irritation and 1 patient needed revision surgery because of implant failure. In the nonoperative group, there were 3 nonunion cases and 2 symptomatic malunions developed requiring corrective osteotomy. At 15 months after intramedullary stabilization, patients in the ESIN group were more satisfied with the appearance of the shoulder and overall outcome, and they benefited a lot from the great improvement of post-traumatic clavicular shortening. Furthermore, DASH scores were lower and Constant scores were significantly higher in contrast to the non-operative group. CONCLUSION: ESIN is a safe minimally invasive surgical technique with lower complication rate, faster return to daily activities, excellent cosmetic and better functional results, restoration of clavicular length for treating mid-shaft clavicular fractures, resulting in high overall satisfaction, which can be regard as an alternative to plate fixation or nonoperative treatment of mid-shaft clavicular fractures.

Osteogenic capability of autologous rabbit adipose-derived stromal cells in repairing calvarial defects.

OBJECTIVE: To evaluate the in vitro and in vivo osteogenic capability of adipose-derived stromal cells (ASCs). METHODS: ASCs were isolated from New Zealand white rabbits and determined by alkaline phosphatase (ALP) staining, von Kossa staining and alizarin red staining. Some specific markers of osteogenic differentiation, including ALP, osteocalcin (OCN), osteopontin (OPN) were examined by reverse transcription-polymerase chain reaction (RT-PCR). In vivo, demineralized bone matrix (DBM)-ASCs composites were implanted into the rabbit calvarial defects created at each side of the longitudinal midline. After 6 weeks, histologic properties of the transplants were analyzed. RESULTS: ASCs were successfully induced into osteogenesis. ALP staining, von Kossa staining and alizarin red staining showed positive results. The expressions of ALP, OCN and OPN were detected in ASCs after cultivation in osteogenic medium. Extensive new bone was observed in the defects transplanted with DBM-ASCs composites. CONCLUSION: ASCs have the potential to differentiate into osteogenic lineage and DBM-ASCs constructs are a promising method for regeneration in bone defects.

Coccygectomy for stubborn coccydynia.

OBJECTIVE: To evaluate the preliminary clinical outcomes of coccygectomy in patients with coccydynia after a failure of conservative treatment. METHODS: From May 2002 to January 2010, 31 patients with coccydynia were treated by coccygectomy in our department after conservative measures had failed to produce significant relief. A questionnaire, which included the extent of relief in the painful area, improvement in quality of life, intensity of pain in the sitting position, and pain score during daily activities, was used to evaluate the results. RESULTS: All patients were followed up for 1 to 6 years (mean 3.3 years). The results were excellent in 20 patients (64.5%), good in 7 patients (22.6%), moderate in 3 patients (9.7%) and poor in 1 patient (3.2%). The excellent and good rates amounted to 87.1%. All patients except one had complete resolution of their symptoms and were subjectively highly satisfied with the outcomes of the surgery. Only 2 cases of superficial infection were observed postoperatively. CONCLUSION: Coccygectomy is a feasible management option for patients with coccygodynia that has no response to conservative treatments.

K-wire and tension band wire fixation in treating sternoclavicular joint dislocation.

OBJECTIVE: To evaluate the feasibility and therapeutic effect of treating sternoclavicular joint dislocation by K-wire and tension band wire fixation, and to improve the safety and stability of this technique. METHODS: This study consisted of 9 cases, 6 males and 3 females with the mean age of 25 years (range, 9-62 years). The causes were traffic accident in 7 cases, falling in 1 case and fight in 1 case. The duration from injury to operation was 2 hours to 7 days. There were 5 left dislocations and 4 right dislocations; 8 anterior dislocations and 1 posterior dislocation, including one combined with left scapular fracture and one with left olecranon fracture. Open reduction and internal fixation using K-wires and tension band wires were performed to treat dislocations. RESULTS: All patients were followed up for 6 to 24 months, 10 months on average. According to Rockwood's rating scale on postoperative sternoclavicular joint, 8 cases achieved excellent outcomes with an average score of 13.88, and the rest case achieved a good outcome with the score of 12. Anatomical reduction was obtained in all cases. There were no such postoperative complications as severe infection, injury to blood vessel and nerve, failure of fixation, etc. Patients were all satisfied with the anatomical reduction and functional recovery. CONCLUSIONS: The technique of K-wire and tension band wire fixation is safe, simple, effective, less invasive and has been successfully used in orthopedic surgery. It is effective in treating sternoclavicular joint dislocation though it has some disadvantages.

[Effects of nicotine on bone marrow stromal cells proliferation and differentiation of chondrocyte in vitro].

OBJECTIVE: To examine the effects of various concentration of nicotine on bone marrow stromal cells (BMSCs) proliferation and differentiation of cartilaginous in vitro. METHODS: BMSCs was obtained from femoral bone and tibia of New-Zealand albino rabbit. The cells of the 3rd generation were used in study. Different concentration of nicotine (0, 1 x 10(-7), 1 x 10(-6), 1 x 10(-5) M) were added into BMSCs. BMSCs proliferation was analyzed by MTT assay at the 1, 4, 7, 14 days. The expression of collagen type II and aggrecan as the marker genes of cartilaginous differentiation from BMSCs were detected by reverse transcriptase-polymerase chain reaction (RT-PCR). RESULTS: Microscope showed that BMSCs transformed from round to fusiform shape. The concentration of nicotine in 1 x 10(-7), 1 x 10(-6) M had a significant positive effect on cell proliferation and the expression of type II collagen in a time-dependent manner when supplemented in commonly used induction media (P<0.05). Concentrations of nicotine in 1 x 10(-7) can promote the expression of aggrecan at the 7th day after induction,and in 1 x 10(-5) M may inhibit the expression of type II collagen and aggrecan. CONCLUSION: It was implied that local application of nicotine at an appropriate concentration may be a promising approach for enhancing cartilaginous differentiation capacity of BMSCs in cartilage tissue engineering.