Methicillin-resistant Staphylococcus aureus ( MRSA) is a common clinic pathogen for nosocomial infections, such as pneumonia, bloodstream infection, endocarditis, skin soft-tissue infection, and osteoarticular infection, which brings giant challenge for clinic treatment.Vancomycin remains an acceptable treatment option, but it needs to be adjusted by pharmacokinetic/pharmacodynamic ( PK/PD ) parameters.Lipoglycopeptides show excellent antimicrobial activity in vitro, but their long half-lives and complex PKs may preclude these agents being used in critically ill patients.Anti-MRSA cephalosporins were reported to be associated with the emergence of its antimicrobial resistance, so clinicians should be cautious when employing these kinds of antibiotics in clinical practice.So far, only linezolid has been proved with better performance than vancomycin for the treatment of hospital acquired pneumonia due to MRSA. Tedizolid, which is also categorized as Oxazolidinone, has higher bioavailability with lower rate of adverse events, but more investigation and validation are still needed for clinic application.Daptomycin displays similar performance with vancomycin on bloodstream infection due to MRSA, so it is recommended as the main drug for the treatment of MRSA associated bloodstream infection.Others such as quinupristin/dalfopristin and tigecycline are all lack of clinical evidence on the treatment of MRSA associated severe infections, so they are only considered when other anti-MRSA drugs showing inferior clinical effects. Rifampicin, Gentamicin, Fosfomycin, Sulfamethoxazole-Trimethoprim and other drugs may be administered for combination therapy, but clinical evidence is still lacking.

Objective To observe the treatment and prophylaxis of deep venous thrombosis(DVT) following fracture of lower limb and the hemorheological changes before and after treatment.Methods 26 patients underwent fracture of lower limb for study received thromholytic therapy,decreasing blood viscosity,promoting blood circulation by eliminating stasis.Before treatment and on the tenth day after treatment,the hemorheological parameters were tested.There existed a statistical analysis for the data processing.Results Among 26 patients,therapeutic evaluation:12 patients was superior,13 patients were favorable,only one patient was infective,and the total effective rate was 96.2%.Curative effect was satisfied.Conclusions The most important was prevention first for the patient with DVT following fracture of lower limb.The critical measure was early diagnosis and early treatment.

BACKGROUND:Transforming growth factor beta-1(TGF-β1)is a cytokine having variously biological effects in repair and renew of tissue injuries; meanwhile, tendon sheath fibroblasts and collagen Ⅰ play important roles in healing and desmoplasia of tendon.OBJECIVE:To study the effects of TGF-β1 on the proliferation and collagen production of tendon sheath fibroblasts.epitenon tenocytes and endotenon tenocytes in the three cell types of rabbit fexor tendon.DESIGN:Contrast observation study.SETTING:Department of Trauma Surgery,Affiliated Hospital of Medical College,Qingdao University.MATERIALS:The experiment was carried out in the Animal Laboratory,Affiliated Hospital of Medical College,Qingdao University from July 2004 to September 2005.A total of 6 adult New Zealand rabbits,of either gender,weighing 3.5-4.5 kg,were selected from Qingdao Experimental Animal Center.Collagenase was provided by Sigma Company;collagen Ⅰ,Ⅱand Ⅲ antibody by Sigma Company;TGF-β1 by Wuhan Boster Biology Company.METHODS: Three cell lines of tendon sheath,epitenon and endotenon were isolated from rabbit flexor tendon and cultured in serum culture media and then in serum-free culture media.In addition,the cells in the experimental group were added with 5 μg/L TGF-β1 in each well,but they were not added with any additive in the control group.MAIN OUTCOME MEASURES:①Proliferation in the two groups was measured with cytometry at 1,2,3 and 4 days after culture.②Preduction of collagens Ⅰ,Ⅱ and Ⅲ was measured with immunohistochemical staining at 4 days after culture.③Collagen contents of the three types were measured with enzyme linked immunosorbent assay(ELISA)in the two groups;expressJon of collagen Ⅰ gene was detected with reverse transcription polymerase chain reaction(RT-PCR).④Contents of collagen Ⅰ induced by TGF-β1 in various dosages of 0,5.10,15 and 20 μg/L were detected with ELISA technique.RESULTS:①Proliferated rates were similar in the two groups at 1 day after culture;however,proliferated rate of tendon sheath fibroblasts was rapidly increased, and there was significant difference as compared with that of epitenontenocytes and endotenon tenocytes(P＜0.05).②Expressions of collagens Ⅰ, Ⅱ and Ⅲ:Immunocytochemical stain demonstrated that three kinds of cells could produce collagens Ⅰ, Ⅱ and Ⅲ;while ELISA indicated that the contents of collagens in three types produced by tendon sheath fibroblasts were the most;in addition,content of collage Ⅰ was higher in the experimental group than that in the control group(P＜0.05-0.01).③Expression of collage Ⅰ gene of tendon sheath fibroblasts was increased as 1.3 times in the experimental group as that in the control group and there was signiflcant difierence(P＜0.01);meanwhile,expressions in epitenon tenocytes and endotenon tenocytes were also higher in the experimental group than those in the control group(P＜0.05).④TGF-β1 in the dosage of 5-10 μg/L had obvious effects on increasing production of collagen;however,production of collagen was not obviously changed when it was affeCted by TGF-β1 in the dosage of 10-20 μg/L.CONCLUSION: TGF-β1 can increase the production of collagen in tendon sheath fibroblasts,epitenon tenocytes and endotenon tenocytes and the expression of collagen Ⅰ gene. In addition, it is important for regulating level of TGF-β1 after tendon injury to prevent adhesion of tendon.

To study the effects of MRF4 transfection on differentiation and expression of myogenic regulatory factors of human rhabdomyosarcoma RD cells, the plasmid-MRF4 cDNA was transfected into cultured rhabdomyosarcoma RD cells with lipofectin method. The myogenic regulatory factors MRF4 and MyoD mRNA were measured with in situ hybridization and the expressions of myosin heavy chain(MHC) and a-actin in the cells were assayed with immunocytochemical method. The cell growth and morphology were observed at the same time. It was found that the morphology of differentiation increased and the growth was suppressed in RD cells after transfection. The expression of MHC and a-actin were significantly increased in RD cells after transfection, while the expressions of MRF4 and MyoD mRNA were up-regulated. It is suggested that transfection of MRF4 can induce differentiation of RD cells and up-regulate the expression of MyoD.

Objective To look for a method for isolation and cultivation of mesenchymal stem cells from human adipose tissue in vitro. Method The adipose tissue was obtained from the omentum of abdominal surgery patients under the aseptic condition. The fat then was minced and digested with 5ml 0 25% trypin for 30 minutes in a 37℃ water bath under constant agitation. The layer with monoclear cells was aspirated and supplemented with albumin, then was cultured in DMEM with 15% calf serum. Immunocytochemistry was used to determine the surface molecule CD44, HLA DR and VWF. Result There was a large amount of mesenchymal stem cells in human adipose tissue. Immunocytochemical staining showed that most of the cultured cells were CD44 positive, few HLA DR positive and VWF positive, indicating that most of the cultured cells were MSC, and the others were fibroblasts and endothelial cells. Conclusion A simple and convenient method to isolate and culture the adipose tissue derived mesenchymal stem cells was successfully established, providing the foundation for the future use of ADMSCs in the treatment of cardiovascular diseases.

BACKGROUND: The skill to culture osteoblasts primarily has been well developed, however, variousdigestive enzymes can affect membrane protein of osteoblasts when they are used on primary tissue separately.OBJECTIVE: To avoid the damage from enzyme to cell as best possible,digest cranial osseous tissue with collagenase in DMEM containing fetal bovine serum and carry out in vitro culture of osteoblasts, then observe the effect of collagenase in DMEM containing fetal bovine serum on the digestion of osteoblasts.DESIGN: Control experiment.SETTING: Department of Orthopaedic Surgery, Tongji Hospital Affiliated to Huazhong University of Science and Technology.MATERIALS: This experiment was carried out in the Department of Orthopaedic Surgery, Tongji Hospital Affiliated to Huazhong University of Science and Technology from March to December 2004. Two Japanese white rabbits with big ears that were pregnant for 28 days were used in the experiment.METHODS: Type I collagenase with the same batch number was prepared into 0.1% collagenase with fetal bovine serum and 0.1% collagenase without fetal bovine serum by using culture medium containing 0.15 volume fraction of fetal bovine serum and DMEM solution respectively, serving as the enzyme digestive juice A and B for experimental group and control group respectively. Abdominal delivery was performed to take ont the fetal rabbit which was at embryonic 28 days under aseptic condition.Then, craniums of fetal rabbits were dissected, rinsed and chipped into pieces. 5 mL solution A and 5 mL solution B were added into the experimental group and control group at 37℃, respectively. Culture solution containing 0.15 volume fraction of fetal bovine serum was gradiently diluted into 0,1,2 and 4 folds in each group, and continued to digest and culture osteoblasts. The cellular survival rate was measured with trypan blue staining, and the osteoblast and its purity were identified with alkaline phosphatase(ALP) staining. Cellular growth was observed under the microscope.MAIN OUTCOME MEASURES: ①Cellular survival rate of two groups was detected with trypan blue staining. ② Cellular purity of two groups was detected with ALP staining in the cells. ③Cellular growth was distinguished by observing cellular morphology under the microscope.RESULTS: ① Results of trypan blue staining: Through cell counting, the rate of cells, which refused to be stained, of the experimental group reached 98%, and the cellular survival rate was high; that of control group reached 95%, and the cellular survival rate was also very high. ② Results of ALP staining in the cells: The area of ALP staining positive of experimental group was not less than 95%, and cellular purity was very high; but that of control group was not more than 75%, and cellular purity was very low. ③ Observation results under the microscope: Cells of experimental group were well stacked, convex and stereoscopic, they grew prosperously and had enough nutrition; while those of control group were not significantly in comparison with experimental group.CONCLUSION: Collagenase containing fetal bovine serum is used to digest the osseous tissue of cranium of fetal rabbits, and the cultured osteoblasts have typical properties of osteoblasts with simple component and high survival rate.

Objective To investigate clinical effects of neoadjuvant chemotherapy(NAC)for phase II B breast cancer.Methods From Jan.2002 to Nov.2004,330 female patients with phase II B unilateral breast cancer were enrolled in the study and they were randomly divided into 2 groups.Group A was NAC group and 152 cased were enrolled in this group,among whom 78 cases had left-side cancer and 74 cases had right-side cancer.The mean age of group A was 42.6 years(ranging from 32 to 73 years).Group A were given beth taxane-and anthracycline- based chemotherapy,according to the protocol of group A.After 4 cycles of chemotherapy,patients in group A underwent conservative breast surgery or radical mastectomy according to the result of chemotherapy.Group B,the non-NAC group,had 178 cases in it,among whom 98 had left-side cancer and 80 had rightside cancer.The mean age of group B was 41.4 years(ranging from 31 to 72 years).Group B also underwent conservative breast surgery or radical mastectomy according to tumor size and axillary node status.According to standards of International Union against Cancer(UICC),the primary focus and axillary node status in NAC group were analyzed before and after chemotherapy.Results Group A had a total effective rate of 91.45%.Breast conservation rate in group A and group B was 44.70%and 21.90%respectively.The overall 3 and 5 year survival rate in group A was 96.05%and 75.00%respectively.which was significantly higher than that in group B.Conclusion NAC is effective in reducing clinical stage of phase II B breast cancer,which increases breast conservation rate and postoperative survival rate.

OBJECTIVE:To optimize ultrasonic-assisted extraction technology of total flavonoids from Vaccinium uliginosum. METHODS:Based on single factor test,the ultrasonic-assisted extraction technology of total flavonoids from V. uliginosum was op-timized by orthogonal test,using the yield of total flavonoids as index,with ethanol volume fraction,solid-liquid ratio and ultrason-ic power as factors. The optimized technology was validated and compared with traditional ethanol reflux method(reflux extracting for 2 times,2 h each time). RESULTS:The optimal extraction technology was as follows as the solid-liquid ration of 1:25,70%ethanol,ultrasonic power of 250 W(40 min for ultrasonic time). In validation test,the yield of total flavonoids from V. uliginosum was 3.98%(RSD=0.41%,n=3),and the yield of total flavonoids was 2.25% by ethanol reflux method. CONCLUSIONS:The ultrasonic-assisted extraction is obviously superior to ethanol reflux method on extraction time and effects,and it is suitable for in-dustrialized production.

Objective To explore the expression of breast cancer susceptibility gene 1 (BRCA1) in human colorectal cancer and its correlation with the efficacy of platinum-based chemotherapy.Methods Seventy-eight cases of colorectal cancer patients treated with platinum-based chemotherapy after surgery were collected.The surgical specimens of them were taken.Fourteen normal colonic mucosa specimens and 12 non-cancerous tissues of colorectal cancer specimens were obtained.The expression of BRCA1 in tissues was detected by immunohistochemistry and analyzed by x2 test.The colorectal cancer patients were followed up for survival time.Comparison of survival analysis was performed by Kaplan-Meier survival curves and Log-rank test.Results The positive rate of the BRCA1 expression in colorectal cancer tissue (52.6 ％,41/78) was lower than that of para-cancer tissue (11/12,x2 =6.518,P=0.011) and normal colonic mucosa tissue (13/14,x2 =7.949,P=0.005).The poorer the differentiation of colorectal cancer,the lower the positive rate of BRCA1 (x2=14.160,P=0.001).The median disease-free survival time of BRCA1 negative colorectal cancer patients was 51.0 months (95 ％ CI:47.7 to 54.4 months),which was longer than that of BRCA1 positive patients (45.0 months,95 ％CI:36.6 to 53.4 months,x2 =4.367,P=0.032).Conclusions Receiving oxaliplatin based chemotherapy may be a survival benefit for BRCA1 negative colorectal cancer patients.The expression of BRCA1 may be an index for chemotherapy options and prognosis evaluation for colorectal cancer patients after surgery.