Objective The mechanism of delaying flap with the minimal invasive surgery was approached to understand its effects on the whole delayed survival of skin flaps thoroughly so as to provide the rationale for its clinical use.Methods 80 male or female Wistar rats were randomly subdivided into two groups:delayed by the minimal invasive surgery,and immediately transfer without delay.Flaps in each group delayed were cut at 1,2,3 and 4 weeks,respectively.The flap was designed at the lower back of rat,with the size of 5 cm × 1 cm,crossing the middle area for 2.5 cm and including stem of iliac branch from iliolumbar artery.2 weeks after second operation,the survival area,capillary density and content of lactic acid of the flaps in each group were examined,and the survival of the falps delayed by the minimal invasive surgery was compared.Results The longer delaying time,and the higher survival rate were observed in the experimental group.Delaying for 3 weeks and 4 weeks,the survival rate was (86.13 ±1.13) ％,(93.49 ± 1.15) ％,respectively,in the experimental group.While in control group,the survival rate was no more than 63％.The longer delaying time,the higher the capillary density were noted in two groups,but 3 weeks delayed group equally matched to the 4 weeks delayed group.In the experimental group,the content of lactic acid increased peaked in 1 week delayed group,then fell-down gradually,but kept steady in 3 and 4 weeks delayed group.The content of lactic acid in the control group kept steady.Conclusions The experimental model is selected as cross-area axial flap on the lower back of rats.The minimal invasive surgery plays the same role as in delaying flaps,which causes vasoconstriction,resulting in disorder of internal environment,ischemia and hypoxia,finally vasodilatation.The more ramus anastomosis,the more survival rate of the flap.

To date,the clustered regularly interspaced short palindromic repeat (CRISPR)/Cas9 system has been widely used to edit the genome in many species and cells.The system is the third generation of artificial endonuclease,which can edit DNA by recognizing short DNA sequences.This paper reviews the structural features of the system and its application in virus research,such as the functional studies of virus-related genes and the exploration of antiviral therapies (including HIV,HBV,and EB virus),looking forward to the future direc-tion of virus research.

Objective To study the effect of mini percutaneous nephrolithotripsy (MPCNL) for non-nephredema kidney calculi. Methods A total of 47 patients with non-nephredema kidney calculi were enrolled in this study. Physiological saline solution was infused into the kidney through a ureteral catheter to induce hydronephrosis. C-arm X-ray machine was employed in percutaneous puncture and Wolf EMS was used to smash the calculi. Results The mean operation time was (120?35) min.The calculi were removed through one passage in 38 cases,through two passages in 6, and three passages in 3. Among the 47 patients, the MPCNL was performed once in 35, and twice in 12. The stone-free rate was 83.0% (39/47) and final stone-free rate was 93.6% (44/47). No complications were noted in all the patients. Conclusions Highly skilled technique is necessary in MPCNL for non-nephredema kidney calculi. The MPCNL is a safe, minimally invasive, and effective method. By using MPCNL, patients have less trauma, lower rate of calculi remaining, and quick recovery. As long as the surgeons are familiar with the technique of MPCNL and the anatomy of non-nephredema kidney, the therapeutic outcome can be as good as that in the patients with nephredema renal calculi.

Objective: To investigate the effects of ?-melanocyte-stimulating hormone(?-MSH) on TGF-?_1 secretion of fibroblasts from keloids so as to find a novel evidence to research etiology of keloids.Methods:Fibroblasts isolated from human keloids were cultured in DMEM medium with 15% bovine serum and treated with ?-MSH. The level of TGF-?_1 was measured with ELISA method. Results: ?-MSH at the concentration of 10 -6 mmol/L triggered the TGF-?_1 secretion of fibroblasts. Conclusion:?-MSH possesses the effect of triggering human keloid fibroblasts to secrete TGF-?_1, and its effective concentration is 10 -6mmol/L.

Objective To analyze the incidence of cerebral vasospasm (CVS) in patients with traumatic subarachnoid hemorrhage(t-SAH), time windows of CVS as well as the risk factors. Method A total of 98 patients,with t -SAH admitted from June 2007 to December 2008, were enrolled for this prospective study. The hemodynamics of middle cerebral artery (MCA) in these patients was monitored with trancranial Doppler (TCD) daily for 7 days after admission and on the 14th day of hospital stay. The incidence of cerebral vasospasm (CVS) in patients with traumatic subarachnoid hemorrhage (t-SAH) ,time windows of CVS as well as the risk factors were analyzed. Results Of them, 41 patients (41.8%) had CVS. The flow velocity of MCA in patients with GCS≤ 8 was significantly higher than that in patients with GCS≥9. Classified by t-SAH cumulative blood Hijdra method, 2(4.44%) of 45 patients(45.9%)with scores 6 or less,9 (29.0%)of 31 patients (37.8%) with scores 6～ 13,and 8 (36.4%) of 22 patients (20.0%)with scores 13 or more had CVS. Severe CVS occurred in 13 (35. 1% )of 37 surgical patients (37.8%), and local cerebral infarction occurred in four surgical patients after symptomatic treatment. The flow velocity of the MCA was significantly higher in surgical patients than that in non-surgical patients 3 days after admission. Conclusions The severity of original trauma, bleeding, location of t-SAH and operation are the major risk factors to lead to CVS in patients with t-SAH. Attention should he paid to those risk factors during the treatment of patients with t-SAH.

AIM:To study pharmacokinetic of Pueraria Flavonid Nasal Drop in rabbits through nasal administration in comparison with oral administration. METHODS:Ten New Zealand rabbits were divided into two groups randomly and administrated nasally and orally (106.4 mg/kg of pueraria flavonid). HPLC was adopted to detect pueraria flavonoid content and DAS 2.0 was used to calculate bioavailability. RESULTS:The main pharmacokinetic parameters of nasal and oral administrations were AUC (0-∞)1 =(30.55?4.93)mg/(L?h),T max1 =(0.90?0.14)h,C max1 =(11.27?1.66)mg/L;AUC (0-∞)2 =(6.90?2.76)mg/(L?h),T max2 =(0.63?0.34)h,C max2 =(1.68?0.84)mg/L. Relative bioavailability of nasal delivery was 442.8%. CONCLUSION:Pueraria Flavonoid Nasal Drop is well absorbed in rabbits and has high bioavailability.

Objective To study the clinical features of invasive pneumococcus disease (IPD) with resistance to antimicrobial agents in children,and to improve the diagnosis and treatment of this disease.Methods The clinical data from 21 IPD patients younger than 13 years old were collected from January 2008 through December 2010 in Pediatric Intensive Care Unit in Beijing Children's Hospital for retrospective analysis. Specimens of blood,pleural effusion,cerebrospinal fluid and soft tissue aspirated were collected from these children,and 23 strains of streptococcus pneumonia (SP) were cultured,isolated and confirmed,and the antibiotics susceptibility to penicillin and other antibiotics of these strains were assayed.Results Among the 21 IPD children,the ratio of male to female was 0.9∶1,and the age was 5 months to 13 years,with 61.9％ of them under 2 years.Of them,12 patients (57.1％ ) had purulent pleurisy,and 1 (4.8％ )patient had an underlying disease diagnosed to be X - linked agammaglobulinemia (XLA).There was no seasonal difference in the occurrence rate of this disease. Eight (38.1％) patients were cured,11(52.4％ ) were improved,while 2 (9.5％ ) patients not improved without death.There was no statistically significant difference in the annual detection rate of invasive SP (x2 =3.711,P =0.156).The incidences of penicillin-intermediate susceptibility SP (PISP) and penicillin-resistant SP (PRSP) were 47.8％ and 26.1％ respectively.The rate of resistance to multiple antibiotics was 91.3％.Conclusions Children aged less than 5 years,especially younger than 2 years,are prone to IPD,and purulent pleurisy and septicemia are often seen in this disease. Some patients had the underlying diseases.The complications included hemophagocytic syndrome,acute respiratory distress syndrome,septic shock,bronchial pleural fistula and so on.The multidrug resistance rate was 91.3％.It is important to put great emphasis on the monitoring antibiotics resistance to invasive pneumococcal disease.

BACKGROUND:Human leukocyte antigen (HLA), the major histocompatibility complex of human, plays an important function in the transplant rejection. Decreasing the expression of HLA wil prolong the survival time of transplants.
OBJECTIVE:To design smal interfering RNA (siRNA) of HLA-A2 and to detect the effect of siRNA-HLA-A2 on the expression of HLA-A2.
METHODS:Four kinds of siRNA-HLA-A2 domains were designed, and recombinant lentivirus expression vector were formed. The 293T cells, highly expressing HLA-A2, were infected in vitro. Then the knockout efficacy of four domains was detected to select the highly efficient siRNA-HLA-A2 target sequences. The human embryo lung fibroblasts were cultured in vitro and infected with the lentivirus carrying the target sequence. The infecting efficiency of LV-siRNA-HLA-A2 was observed under the fluorescence microscope and the silence function of this siRNA in human embryo lung fibroblasts was detected by western blot analysis.
RESULTS AND CONCLUSION:According to the mRNA sequence of HLA-A2 in Genbank, three siRNAs were designed and synthesized. In vitro, the over expression of HLA-A2 in 293K cells was successful y silenced. The HLA-A2 expression in human embryo lung fibroblasts was also efficiently silenced after the human embryo lung fibroblasts were infected by the highly efficient siRNA of HLA-A2. The efficacy was up to 80%.

10.3969/j.issn.2095-4344.2013.25.010

Objective To explore the safety of the human bone marrow‐derived mesenchymal stem cells (hBMSCs) after silencing of human leukocyte antigen A2 expression .Methods We divided the cells into three groups:normal cultured cells of the 8th passage served as control group , and hBMSCs after HLA A2 silencing expression of the 5th and 15th passage as experimental groups 1 and 2 ,respectively .The hBMSCs were recultured by sterile methods .The growth curve ,telomerase activation ,and expressions of P27 ,cyclin D2 and cyclin‐dependent kinase 4 (CDK4) were utilized to explore the safety of the hBMSCs induced by LV‐siRNA‐HLA A2 .The BMSCs were transplanted to the subcutaneous layer of nude mice .Tissue types were detected 24 weeks after transplantation . Results The cell curves had no obvious left or right shift in all the groups . The telomerease activation in experimental groups 1 and 2 did not significantly differ from those in control group . The expressions of anti‐oncogene P27 ,cyclin D2 and CDK4 had no obvious difference between the two experimental groups and control group , either . There was only ectopic osteogenesis 24 weeks after the BMSCs (HLA A2 gene silenced ) were transplanted to the subcutaneous layer of the nude mice .Conclusion There was no obvious evidence to support that hBMSCs had undergone change in safety after the silencing of HLA A 2 expression .