AIM To establish an HPLC method for the simuhaneous content determination of five flavonoids in Duchesnea indica (Andr.) Focke at five picking time (April,May,June,July and August).METHODS The analysis of D.indica methanol extract was performed on a 25 ℃ thermostatic Agilent ZORBAX SB-C18 column (250 mm×4.6 mm,5 μm),the mobile phase comprising of acetointrile-0.1％ methanoic acid flowing at 1.0 mL/min in a gradient elution manner,and the detection wavelength was set at 350 nm.RESULTS Rutin,hyperoside,isoquercitrin,celereoin and kaempferol showed good linear relationships within their own ranges (R2 ≥0.998 6),whose average recoveries were 97.1％-101.5％ with the RSDs of 1.37％-2.37％.The contents of five constituents in samples at different picking time exhibited obvious differences,among which lutin and hyperoside contents were the highest in June,isoquercitrin content was the highest in July,and celereoin and kaempferol contents were the highest in August.CONCLUSION The suitable picking time of D.indica is June and July.

Objective To study the changes in the expressions of CXCR4 of leucocytes as modulated by erythrocytes from cord blood. Methods 0.2ml suspension of whole blood cells (or 0.2ml suspension of leucocytes) was added to 0.3ml of auto plasma, and 0.2ml suspension of S180 (5?10~6/ml) was added as stimuli. The suspension was incubated at 37 ℃ for 1 hour. Normal saline instead of S180 was used as control The expression changes in CD35 on erythrocytes and CXCR4 on leucocytes were determined with flow cytometry assay. The data could be grouped under four heads: (1) cancer cells 0.2ml were added to whole blood cells 0.2ml and plasma 0.3ml; (2) NS 0.2ml were added to whole blood cells 0.2ml and plasma 0.3ml; (3) cancer cells 0.2ml were added to white blood cells 0.2ml and plasma 0.3ml; (4) NS 0.2ml were added to white blood cells and plasma 0.3ml. Results When activated by S180, the expression of CD35 on cord erythrocytes was decline not so low as that of adult′; while the expression of CXCR4 on cord leucocytes was increased not so high as that of adults′. When activated by S180, the expression of CD35 on adult erythrocytes was significantly decreased than that of group of not activating (40.21%?11.52% and 28.65%?8.08% respectively), while the expression of CXCR4 on adult erythrocytes was significantly increased (40.62%?17.89% and 64.18%?11.69% respectively, P

Objective To evaluate of red blood cell as giving instruction in whole white blood cell immunological activity by new nature experimental system of hemaimmune reaction rood map. Methods Plasma 0.3ml were added to whole blood cells (including: red blood cell and white blood cells) or white blood cells 0.2ml, and incubated for 1h at 37℃. The content of IL-8 and IL-12 was determined by enzyme linked immunadsorbent assay (ELISA) method. The expression level of CD4, CD8, CD35 and CXCR4 on white blood cells was determined by method of Flow Cytometry. Results The content of IL-8 (5.96?4.26) and IL-12 (9.84?2.23) in whole blood and plasma nature group was significantly lower than that (13.59?3.69?B?pg~ -1 ?ml~ -1 ) and (15.09?9.86?B?pg~ -1 ?ml~ -1 ) in white blood cell and plasma isolation group (P

Objective To investigate the value of prenatal ultrasound in diagnosis and treatment of twin reverse arterial perfusion (TRAP) syndrome. Methods A retrospective study was performed in 5 TRAP cases, including ultrasound images, clinical data and pregnancy outcomes. The sonographic characteristics were summarized. Results Five TRAP cases were diagnosed during 13 to 28 weeks' gestation and confirmed after birth. Color Doppler unltrasonography revealed retrograde umbilical artery perfusion towards acardiac twin. Two of 5 cases ended up in induced abortion, 1 in spontaneous abortion, 1 was delivered at 37 weeks' gestation after ultrasound-guided feticide of the acardiac twin and 1 was monitored closely with ultrasound and delivered alive at 32~(+4) weeks' gestation. Conclusion Prenatal ultrasonography has great applicative value for TRAP syndrome in early diagnosis, choosing optimal treatment and prognosis assessment.

Shiga toxin 2 (Stx2) toxoid produced by formaldehyde treatment of purified toxin was used to immunize BALB/c mice for monoclonal antibody (MAb) production.The neutralizing activities of positive clones against Stx2 were screened by in vitro cytotoxicity assay.The isotype and specificity of resultant clone was determined,and its efficacy to neutralize the activity of purified Stx2 was evaluated by in vitro and in vivo toxicity model.Lastly,its spectrum of activity against Stx2 variants was also accessed by mouse toxicity model.It was demonstrated that one of the 12 positive MAb clones against Stx2,designating $2C4 had neutralizing activity.S2C4 belongs to the immunoglobulin G1 subclass and has a K light chain,and it reacts with the A subunit of Stx2 and does not bind to Stx2 B subunit or to Stx1.S2CA could efficiently neutralize the cytotoxicity of Stx2 to Veto cells and mice.It also protected mice against lethal doses of Stx2 variants challenge including Stx2c and Stx2vha.S2C4 is a promising candidate molecule in preventing the progression of hemolytic-uremic syndrome (HUS) mediated mainly by Stx2 in Stx-producing Escherichia coli(STEC)infection.

OBJECTIVETo investigate the hemodynamics evidence of the descending branch of lateral circumflex femoral artery in a reversed way. To explore the clinical result of using the reversed descending branch of the lateral circumflex femoral artery as the receipt artery for free flaps for reconstruction of the leg soft-tissue defect.

METHODSFrom October 2005 to February 2012, 38 patients with severe leg soft-tissue defects were treated. The proximal antegrade and retrograde mean artery pressure of the descending branch of the lateral circumflex femoral artery in 16 of 38 patients were recorded during operation. All wounds had osteomyelitis, bone and tendon exposure requiring coverage reconstruction. And there was no recipient artery in the injured lower leg for free flaps in all 38 patients. Reversed descending branches of lateral femoral circumflex arteries were used as recipient arteries for free flaps (free latissimus dorsi flap, free thoracoumbilical flap, and free anterolateral thigh flap) in all patients. The flap donor site was closed directly or with the skin graft.

RESULTSThe proximal antegrade mean artery pressure of the descending branch of lateral circumflex femoral artery was(81.6 +/- 12.4) mmHg. The proximal retrograde pressure was(48.2 +/- 10.7) mmHg. The proximal retrograde mean artery pressure was 59.07 percent of the proximal antegrade pressure. The donor skin graft survived and wound healed primarily. After operation, 2 flaps had distal partial necrosis and healing was achieved after dressing change. All the other flaps survived completely without vascular problems. All the patients were followed up for 11 months to 2.5 years (mean, 1.6 years). The flap appearance was satisfactory. The texture and color of flaps in all cases were good.

CONCLUSIONSThe reverse descending branch of lateral circumflex femoral artery is a reliable recipient artery for the free flaps. It is an easy and simple technique that can be used for reconstruction of the defects in the lower leg, with the reversed descending branch of lateral circumflex femoral artery as recipient artery.

Adolescent ; Adult ; Blood Pressure ; Female ; Femoral Artery ; physiopathology ; surgery ; Free Tissue Flaps ; blood supply ; Hemodynamics ; Humans ; Lower Extremity ; injuries ; Male ; Middle Aged ; Soft Tissue Injuries ; surgery ; Young Adult

Objective To purify Shiga toxin Ⅱ (STX Ⅱ) of enterohaemorrhagic Escherichia coli (EHEC) O157: H7 by affinity chromatography, and characterize its biological function. Methods The immno-affinity chromatography column was prepared by STX Ⅱ A subunit-specific antibody S1D8 coupling to Sepharose 4B matrix. The purity and specificity of STX Ⅱ molecule secreted by EHEC O157:H7 were detected by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and Western blot, respectively. The purified toxin was serially diluted and the toxic activities to Vero cell line and mice were observed. The 50% cytotoxic dose (CD50) for Vero cell line and 100% lethal dose (LD100) for mice were calculated. The protection effect of anti-STX Ⅱ polysera to the mice against the purified toxin challenge was also observed. Results STX Ⅱ was successfully purified from culture supernatant of EHEC O157:H7 using affinity chromatography scheme. The relative molecular weights of STX Ⅱ A and B subunits were 32 000 and 7 500 confirmed by SDS-PAGE, respectively. The purified toxin could react with monoclonal antibodies against STX Ⅱ A and B subunits, respectively.The toxin was cytotoxic to Vero cell with CD50 of 20 ng/L and lethal to mice with LD100 of 5 ng.The toxin could be neutralized by anti-STX Ⅱ polysera in vivo. Conclusion STX Ⅱ is successfully purified and its toxic effects are confirmed in both cell line and mouse model.

Objective:To explore the benefits of speech training for young persons with tongue cancer after 125Ⅰ radioactive seed implantation. Methods:Fifty young persons bearing a 125Ⅰ radioactive seed implanted to treat tongue cancer were given up to 40min of daily tongue function training beginning on the second day after the operation. Abnormal sounds were corrected in the seven days before their discharge. An online platform assisted further training in the family combined with biofeedback. Daily 20-30 minute sessions were prescribed. Time domain waveforms, speech spectra and energy waterfalls were recorded from the 50 subjects before the experiment and after 1 week of the speech training. Intelligibility (PI) was quantified before the operation and the speech training, and then 1, 2, 4 and 6 months after discharge. Results:After 1 week of speech training abnormal waveforms had disappeared from the time domain waveform diagram. Fricative disturbance areas in the speech spectrum diagram had improved significantly, and the energy distribution in the energy waterfall diagram was balanced, showing a trend typical of normal speech fluctuation. The average PI before the operation (39.92±8.46) increased to (40.32±8.79) before the speech training and to (42.78±8.24) one month after discharge. It then continued to improve to (72.32±5.55) 6 months after discharge.Conclusion:Speech training can significantly improve the speech intelligibility of patients with tongue cancer after 125Ⅰ radioactive seed implantation.

Although most microbes are not readily cultured in the lab, microbial DNA can be extracted directly from an environmental sample and be functionally expressed in a suitable host for natural products discovery, and this approach has been termed "metagenomics". An E'mei Mountain soil metagenomic library was constructed using an Escherichia coli-Streptomyces shuttle vector for functional based screening of anti-bacterial clones in Streptomyces albus host. Two active clones were obtained and their fermentation broths were studied for the inhibitory effect on Staphylococcus aureus biofilm. Their fermentation products have a good inhibitory effect on the formation of S. aureus biofilm, and the inhibitory effect could exceed 90% when the concentration of sample was 2 MIC (Minimum Inhibitory Concentration). In addition, two samples had significantly effect on S. aureus biofilm dispersal, and the clearance rate of EM110 was higher than EM123. In conclusion, substances with strong bioactivities on biofilm formation and dispersal of S. aureus could be discovered by using metagenomics technology.

AIM:Bone morphogenetic protein 7(BMP7)reduces the expression of Yes-related protein 1(YAP1)by down-regulating Ajuba level and decreasing extracellular matrix(ECM)deposition.This study aimed to inves-tigate the influence of these factors on modifying the degree of renal fibrosis in rats with diabetic nephropathy.METH-ODS:Eighteen Sprague-Dawley(SD)rats were randomly divided into three groups:the normal control(NC)group,the diabetes mellitus(DM)group,and the DM group treated with BMP7 overexpressing adeno-associated virus(DM+rAAV-BMP7).Each group consisted of six rats.Diabetic kidney disease(DKD)was established in the DM and DM+rAAV-BMP7 groups by injecting 55 mg/kg streptozotocin(STZ)via the tail vein.NRK-52E cells were divided into three groups:the normal glucose(NG)group,the high glucose(HG)group,and the high glucose group treated with recombinant hu-man BMP7(HG+rhBMP7)group.Pathological changes in renal tissues were observed using hematoxylin and eosin(HE)and Sirius red staining.Immunohistochemical staining was performed to examine the expression sites of Ajuba and YAP1 in the renal cortex.Western blot analysis was conducted to determine the expression levels of BMP7,Ajuba,YAP1,colla-gen type Ⅲ(Col-Ⅲ),and fibronectin(FN)in the rat renal cortex and NRK-52E cells.RT-qPCR was used to measure the mRNA levels of Ajuba and YAP1 in the rat renal cortex.RESULTS:Biochemical indices revealed significantly ele-vated levels of blood glucose,serum creatinine,triglycerides,total cholesterol,and 24-hour urinary protein in the DM group compared to the NC group(P<0.05).In the DM+rAAV-BMP7 group,the levels of serum creatinine,24-hour uri-nary protein,triglycerides,and total cholesterol were lower than those in the DM group(P<0.05).Pathological staining demonstrated that the renal interstitium of the DM group exhibited inflammatory cell infiltration,fibrous tissue,collagen fi-ber deposition,disordered renal tubule arrangement,atrophy,and vacuolar degeneration,which were ameliorated in the DM+rAAV-BMP7 group.Immunohistochemistry revealed that Ajuba and YAP1 were mainly expressed in the cytoplasm and nucleus,with high expression in the cytoplasm of the DM group,which was significantly decreased in the DM+rAAV-BMP7 group.Western blot results indicated that the protein levels of FN,Col-Ⅲ,Ajuba,and YAP1 were up-regulated in the DM and the HG groups(P<0.05),but significantly down-regulated in the DM+rAAV-BMP7 group(P<0.05).RT-qP-CR results demonstrated that the mRNA levels of Ajuba and YAP1 were higher in the DM group and significantly lower in the DM+rAAV-BMP7 group(P<0.05).CONCLUSION:The overexpression of BMP7 can ameliorate renal fibrosis in rats with DKD.This effect is likely mediated by the down-regulation of Ajuba,reduction of YAP1 expression,and subse-quent inhibition of ECM deposition.