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【Objective】 To use hairy enhancer of split 1 (Hes1) to regulate the differentiation of liver epithelial progenitor cells (LEPCs) into cholangiocytes. 【Methods】 The vectors, pTet-on and pTRE2hyg-Hes1, were transfected into LEPCs. The expression of Hes1 was induced by doxycycline (DOX) with different concentrations (0, 0.1, 1, 5, 10, 50, 100 and 500 μg/mL). The expressions of Hes1, molecular markers of hepatocyte and cholangiocyte, glutathione synthetase (Gss), keratin 19 (Krt19) and hepatic nuclear factor 1β (HNF1β) in LEPCs were verified by Western blotting, RT-PCR, Real-time PCR, immunocytochemistry and immunofluorescence. 【Results】 The expression of Hes1 in LEPCs transfected by pTet-on/pTRE2hyg-Hes1 was increased by 11.21 fold when induced by DOX at 10 ug/mL, which drove the LEPCs to differentiate into biliary epithelial cells. With increasing expression of Hes1, cholangiocyte markers, Krt19 and HNF1β, were significantly upregulated, while the hepatocyte marker, Gss, was obviously downregulated. 【Conclusion】 DOX at 10 μg/mL may induce a suitably up-regulated expression of Hes1 in LEPCs double-transfected by pTet-on and pTRE2hyg-Hes1, and the suitable high-expression rather than over-expression of Hes1 can regulate LEPCs to differentiate into cholangiocytes.
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Based on the analysis of the theoretical calculation model of axial resolution of optical coherence tomograph for the posterior segment of the human eye, a set of testing device for measuring its axial resolution is designed and developed. In view of a commercial ophthalmic optical coherence tomograph in clinical use, its axial resolution is calculated to be 5.07 μm theoretically, and the actual measurement value is 5.45 μm. The uncertainty of the detection device is evaluated and the result is (5.45±0.10) μm. The measurement error introduced by the testing device is very small. Meanwhile, the axial resolution measured by the testing device meets the requirements of the instrument(≤ 6 μm).
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Humans , Algorithms , Eye , Diagnostic Imaging , Tomography, Optical Coherence , UncertaintyABSTRACT
Based on the Gullstrand I model eye, a simplified model eye for testing fundus imaging device is designed. The model eye can reach the following requirements:(1) The refractive characteristics of the ocular refractive tissue are simulated, and the equivalent focal length in air is 17 mm; (2) The differences between relative refractive index differences of the adjacent materials of the simplified model eye and relative refractive index differences of any adjacent two layers (cornea and aqueous humor, aqueous humor and lens, lens and vitreous body) of the Gullstrand I model eye are not more than 1%; (3) In the case of the incident aperture diameter of 3 mm, the differences of radii of the diffuse spots formed by the paraxial light and the axial light are not more than 15%; (4) The differences of angles of chief ray and tangent line of the fundus are not more than 1°; (5) In the case of the incident aperture diameter of 3 mm, the differences of MTF values of the near axis light are not more than 0.1. The simplified model eye can be expected to be used for testing fundus imaging device instead of the test method in ISO 10940:2009 Ophthalmic instruments-Fundus cameras.
Subject(s)
Cornea , Fundus Oculi , Lens, Crystalline , Diagnostic Imaging , Refraction, OcularABSTRACT
<p><b>OBJECTIVE</b>To observe if VIR576, an 20-mer peptide derived from the C-proximal subfragment of a1-antitrypsin (a1-AT) which inhibits human immunodeficiency virus type 1 (HIV-1) entry into the target cells by interacting with fusion peptide (FP), can also directly inhibit CD4(+) T cell activation in vitro.</p><p><b>METHODS</b>Splenocytes isolated from DO11.10 OVA Tg mice were stimulated with ovalbumin or concanavalin A to test the effects of VIR576 on antigen-specific or non-antigen-specific T cell activation. Both primary CD4(+)CD25(-) T cells from DO11.10 mice and CD4(+) T cell line A2b were activated with specific antigens to evaluate the effects of VIR576.</p><p><b>RESULTS</b>VIR576 inhibited antigen-specific splenocyte activation but had no significant effect on non-antigen-specific T-cell activation, which bypassed the crosstalk between the CD3-signaling complex and TCR. We furthermore observed that VIR576 could also down-regulate antigen-specific CD4(+) T-cell activation.</p><p><b>CONCLUSIONS</b>Given the high susceptibility of activated CD4(+) T cells in the mucosa to HIV-1 infection, the inhibitory effects of VIR576 on both HIV entry into the target cells and CD4(+) T-cell activation suggest the potential of VIR576 as a microbicide for prevention of sexual transmission of HIV.</p>
Subject(s)
Animals , Mice , CD3 Complex , CD4-Positive T-Lymphocytes , HIV Fusion Inhibitors , Pharmacology , HIV-1 , Lymphocyte Activation , Mice, Transgenic , Ovalbumin , Peptide Fragments , Pharmacology , alpha 1-Antitrypsin , PharmacologyABSTRACT
Different from the previous knowledge regarding reactive oxygen species (ROS), recent research suggests that ROS play essential roles in initiating cascade reaction of wound healing. During wound healing, ROS can serve as the second messenger to regulate signal transduction and gene expression. In this paper, we review the mechanism of generation of cyclooxygenase 2-prostaglandin E2 induced by ROS, which regulates the early inflammatory response and subsequent healing after injury.
Subject(s)
Cyclooxygenase 2 , Metabolism , Dinoprostone , Metabolism , Reactive Oxygen Species , Metabolism , Reverse Transcriptase Polymerase Chain Reaction , Methods , Signal Transduction , Wound Healing , PhysiologyABSTRACT
Objective To compare the efficacy of different volume of ropivacaine and lidocaine for ultrasound-guided infraclavicular brachial block.Methods Sixty patients scheduled for distal upper limb surgery were divided into group A and group B by random digits table method with 30 cases each.The patients were given ultrasound-guided infraclavicular brachial block,but the patients in group A using 0.3% ropivacaine mixed with 1% lidocaine 20 ml,the patients in group B using the same mixture 30 ml.Sensory block was assessed every 5 min after the block.Motor block was evaluated at 30 min after the block.The onset time of anesthesia,surgical success rate,complications and duration of anesthesia were recorded.Results There was no significant difference in the surgical success rate [93.3%(28/30) vs.96.7%(29/30),P>0.05],onset time of sensory block [10 (5) min vs.5 (5) min,P>0.05] and degree of motor block between the 2 groups.The duration of anesthesia in group A was shorter than that in group B (P < 0.05).Conclusion The efficacy of ultrasound-guided infraclavicular brachial block with the mixture of 0.3% ropivacaine and 1% lidocaine 20 ml and 30 ml is similar in surgical success rate and onset time of anesthesia.
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Objective To compare the clinical effect of remifentanil and dexmedetomidine on controlled hypotension during endoscopic sinus surgery. Methods Forty patients undergoing endoscopic sinus surgery were randomly divided into group R and group D, 20 in each group. All patients received controlled hypotension when the surgeons began to sterilize the nasal cavity for the purpose of maintaining the MAP at 60 ~ 70 mmHg. Group R received remifentanil 1μg/kg over 1 minute, followed by 0.2 to 0.4μg/kg per minute infusion during maintenance, whereas group D received dexmedetomidine 1μg/kg over 10 minutes, followed by 0.2 to 0.7μg/kg per hour during maintenance. At the moment of pre-induction(T0), incision(T1), 30 min(T2) and 60 min(T3) after incision and 10 min(T4) after end of controlled hypotension, MAP and HR were measured. Quality of the surgical field, blood loss and extubation time were recorded. Sedation and postoperative pain were assessed on arrival at the PACU. Results The goals for the MAP level were achieved and the surgical field quality was ideal in both groups. The MAP and HR values at T1~ T3 were significantly lower than those at T0 in both groups (P 0.05). No significant difference was founded between the groups in the Fromme operative field score and blood loss (P > 0.05). The extubation time was significantly shorter in group R than in group D(P<0.05). The sedation score in the PACU was significantly lower in group R than in group D(P<0.05), while there were no significant differences in postoperative pain between the groups(P<0.05). Conclusion Both remifentanil and dexmedetomidine are safe agents for controlled hypotension and are effective in providing satisfactory surgical fields during endoscopic sinus surgery. However , remifentanil is advantageous in time for extubation and complete consciousness from anesthesia and therefore it may have the advantage of fast track anesthesia.
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Objective To compare the effect between i-gel mask and SHPA mask in the airway management of general anesthesia.Methods Sixty patients of general anesthesia with ASA I or Ⅱ were randomly divided into two groups:i-gel mask group (30 cases) and SLIPA group(30 cases).After intubation and general anesthesia,fiberoptic bronchoscopy was used to detect and evaluate.The peak airway pressure,mean airway pressure,condition of the laryngeal mask intubation,time of intubation,time of extubation,time of recovery and the side effect occurrence of extubation were recorded.Results The time of anesthesia,time of operation,time of extubation,time of recovery,achievement ratio of first time,achievement ratio of second time,time of intubation,peak airway pressure and mean airway pressure had no significant difference between two groups (P > 0.05).The airway seal pressure in i-gel mask group was significantly higher than that in SL1PA group [(29 + 6) cm H2O (1 c m H2O =0.098 kPa) vs.(23 ± 4) cm H2O] (P < 0.05).For the grade of fiberoptic bronchoscopy in i-gel mask group,there was 27 patients in I grade,3 patients in Ⅱ grade.But in SL1PA group,there was 3 patients in I grade,4 patients in Ⅱ grade,13 patients in Ⅲ grade,10 patients in ⅣVgrade.There was significant difference between two groups (P < 0.05).There were none of regurgitation in two groups.Conclusions The i-gel mask and SLIPA mask can provide the same effective ventilation.The i-gel mask has higher airway seal pressure,less side effect during the intubation and extubation,and have better airway management of general anesthesia.
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Extracellular matrix (ECM)is not only one of current hot-points in medical cell biology,but also one tricky part for undergraduates to learn. This article compared chapters of ECM in medical cell biology courses and spotlighted that ECM chapter was often neglected in some domes-tic universities. Then it analyzed the possible causes,such as variable arrangement of ECM section in current textbooks. Lastly,the article recommended several suggestions,including giving a timely re-vision of the old ECM knowledge,designing an appropriate strategy for teaching,enumerating certain representative diseases to improve the ECM education. It appealed our teachers to pay more attention to how to make the module of ECM master well in near future.
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ObjectiveTo assess the effects of fentanyl (F),alfentanil (A),sufentaril (S),butorphanol (B),dezocine (D) and pentazocine (P) on human sperm motility in vitro.MethodsTwenty samples of semen with normal sperm motility were collected.The semen was mixed with HTF liquid culture medium and sperm concentration was adjusted to (40-80) × 106/ml.Then semen suspension 50 μl was added into the culture dishes.The semen suspensions were randomly divided into 3 groups:group Ⅰ control-semen suspension 50 μl + 0.9% NaCl 50 μl.GroupⅡ single opioid-semen suspension 50 μl + 50 μl of 4 concentrations of F,A,S,B,D and P (10-3,10-4,10-5,10-7 mol/L).Group Ⅲ combined opioid-naloxone-semen suspension 50 μl + 25 μl of 3concentrations of naloxone (2 × 10-8,1 × 10-7,2 × 10-7 mol/L) + 25 μl of 4 concentrations of F,A,S,B and D.After being incubated for 1 h the sperm motility was assessed by sperm analyzer and the amplitude of change in sperm motility was calculated.Median inhibition concentration (IC50) and complete inhibition concentration ( IC100 ) of the 6 opioids were calculated from concentration-response curve.Results( 1 ) The shape of concentration-response curve for F,A and S was straight line.The maximal decrease in sperm motility was (56 ± 5)%(F),(58±7)% (A) and(79±:6)% (S).(2) The shape of concentration-response curve for B,D and P was sigmoid.Low concentrations of B,D and P did not affect sperm motility.Median and high concentrations of B and D could inhibit sperm motility.The maximal inhibition was 100%.High concentration of P increased sperm motility.The maximal increase was (19 ± 6)%.(3) Naloxone shifted the concentration-response curve for F,A,S,B and D to the right,and decreased the maximal inhibitory effect of F,A and S dose-dependently but did not affect the maximal inhibitory effect of B and D.(4) IC50 was in the order of A≈ F > D > S≈- B.IC100 was D > B.ConclusionF,A and S inhibit sperm motility by acting on opioid receptor on human sperms while B and D act through opioid or nonopioid pathways.P can increase sperm motility.
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Objective To evaluate the effect of the expression of P-glycoprotein (P-gp) in tumor tissue on analgesic efficacy of morphine and buprenorphine in advanced cancer patients with pain. Methods One hundred and fifty advanced cancer patients with pain aged 51-64 yr weighing 54-65 kg were included in this study. The expression of P-gp was negative in the tumor tissue in 50 patients (group M1 and B1, n = 25 each) and positive in 100 patients (group M2 ,M3 ,and B2 ,B3, n =25 each). The PCA regimen for the 6 groups were listed in the table .Pain was assessed with VAS scores (0 = no pain, 10 = worst pain) and venous blood samples were taken for determination of blood morphine/buprenorphine concentrations at 4, 12, 24 and 48 h of PCIA. Results The six groups were comparable with respect to age, body weight, M/F sex ratio, types of cancer, baseline pain level and education. The analgesic efficacy of morphine and buprenorphine was better ( VAS scores were significantly lower)in P-gp expression negative patients (group M1 and B1 ) than in P-gp expression positive patients (group M2 and B2 ). Higher doses of morphine and buprenorphine provided better analgesic efficacy in P-gp expression positive patients in group M3 and B3 than in group M2 and B2. Plasma morphine and buprenorphine concentrations were comparable between group M1 , B1 and M2, B2 and were significantly higher in group M3 and B3 at each time point. Conclusion Positive P-gp expression in the tumor tissue can decrease the analgesic efficacy of morphine and buprenorphine in advanced cancer patients with pain.
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Objective To study the application of Supreme laryngeal mask airway (SLMA) with new type to the general anesthesia of lateral position. Methods Sixty patients with ASA Ⅰ - Ⅱ who planned to receive selective poster laparoscopic surgery were divided into SLMA group and general tracheal tuber (GTT) group with 30 cases of each by random digits table. After fast intravenous induction SLMA and GTT were intubated respectively to obtain artificial airway in each group. The time of intubation, efficiency of success,heart rate(HR), invasive blood pressure(IVBP),airway pressure,side effects were observed and recorded. Results The intubation time in SLMA group [(15.1 ±4.0) s] was shorter than that in GTT group [(30.5 ±: 16.2) s] (t' =-5.05,P <0.05); there was no significant difference in efficiency of success between two groups [96.7%(29/30) vs. 76.7%(23/30),P> 0.05]. There was no significant difference in HR and IVBP during the time of intubation and extubation in SLMA group(P> 0.05), but not in GTT group(P<0.05). There was no significant difference in the change of the airway pressure between two groups during the ventilation. There were no complications in two groups. Conclusions The time of SLMA intubation in the general anesthesia with lateral position is short. The effect of intubation and extubation to HR and blood pressure is light. The SLMA intubation is another way to set up safely artificial airway.
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Aim: To investigate the pathological changes in NTBC[2-(2-nitro-4-trifluoro-methyl-benzoyl) -1,3 cy-clohexanedione]-induced hepatic injury in mice and in the repopulation of adult hepatocytes in Fah~(-/-) mouse. Methods: Autogenous hepatic injuries in Fah~(-/-) mice were induced by the treatment of NTBC. Injection of hepatocytes obtained from wild-type mice to spleen were transplanted into the Fah~(-/-) mice. Then, changes to body weight and the likelihood of the transplanted Fah~(-/-) mice, and hepatic immunohistochemistry were ob-served. In addition, pathological changes to liver damage induced by NTBC treatment were analyzed under HE-staining microscopy and electron microscopy. Results: The surviving Fah~(-/-) mice subjected to hepatocyte trans-plantation were found to be healthy and in stable body weight. liver repopulation reached to 90% in the 8th week. Repopulating hepatocytes caused no alteration to histological structure of the recipient liver, and subacute hepatic injury occurred in the Fah~(-/-) mice after NTBC treatment. Electronic microscopy observations indicated that necrosis in the hepatocytes occurred at early stage and that apoptosis gradually appeared. It was also shown both necrosis and apoptosis co-existed in the same samples of interest at the following stages of the induced liver injury. Conclusion: Transplanted hepatocytes proliferated in Fah~(-/-) mice allow 90% of the hepatocytic repopula- tion. Repopulation renders normal hepatic function and structure in the recipient Fah~(-/-) mice, as a model of liver repopulation, could be applicable in study of stem cell derived hepatic cells in transplantation assay.
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A computer-based online search of PubMed database and Chinese Journal Full-text Database was undertaken to identify articles about the monitoring of pulmonary arterial pressure (PAP),mixed venous oxygen saturation (SvO2),and breathing mechanics in the patients undergoing lung transplantation from January 1990 to May 2008 to evaluate their clinical values in maintaining cardiorespiratory function of patients during anesthetic stage. PAP,cardiac output and SvO2 are key parameters of monitoring during the diseased lung resection and donor lung transplantation; breathing mechanics and SvO2 are key parameters of monitoring after lung ventilation. The former are reliable indicator to predict lung transplantation reaction and pulmonary edema and the latter can guide regulation of positive end expiratory pressure and when to remove extracorporeal membrane oxygenator. A combination of hemodynamics (PAP,pulmonary capillary wedge pressure,cardiac output,and central venous pressure),breathing mechanics and SvO2 consists an overall monitoring of heart and lung function. By Meta-analysis,these indicators can guide the regulation and protection of heart and lung function in patients undergoing lung transplantation and other severe patients during the anesthesia period.
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Objective: To improve the micro-manipulation of mouse fertilized eggs,promoting its survival rate and productivity in pseudo-pregnant mice. Methods: DNA fragments of definite concentration and size were used for micro-manipulation with conventional and improved methods, then their survival rate of manupulated eggs and productivity of pseudo-pregnant mice were analyzed and compared. Results: With improved methods, the survival rate of eggs increased from 60.3% to 79.5%, the productivity of pseudo-pregnant mice and offspring increased from 35.7% to 56.8% and 10.1% to 14.1% respectively. Conclusion: The improved methods of micro-manipulation can be used in transgenic mouse production.
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Objective: To clone mouse rod opsin promoter (ROP) and establish transgenic mice harboring mouse rod opsin promoter and enhanced green fluorescent protein(mROP-EGFP) fusion gene. Methods: Mouse ROP was cloned from C57BL/6 mouse genomic DNA by polymerase chain reaction (PCR). Expression vector of mROP-EGFP fusion gene were constructed by recombination DNA technique. It was identified by restriction endonucleases digestion and confirmed by DNA sequencing. After Not I restriction endonuclease digestion, the coding elements were microinjected into male pronuclei of mice zygotes to generate transgenic mice. The pups were evaluated by PCR at genomic DNA level and mated with normal mouse. Expression of GFP in retina of transgenic mice was detected by fluorescent microscope. Results: 2. 1 kb mouse rod opsin promoter fragment was amplified from mice genome DNA. Expression vector pmROP-EGFP was constructed successfully. Following microinjection of coding sequence of pmROP-EGFP, 3 pups were verified to integrate the mROP-EGFP fusion gene in their genomic DNA by PCR assay, named C57-TgN (mROP-EGFP )SMMU21, C57-TgN (mROP-EGFP)SM-MU26 and C57-TgN(mROP-EGFP) SMMU27. They could express GFP in retina. Conclusion: 2. 1 kb mouse rod opsin promoter is cloned and expression vector pmROP-EGFP is constructed. mROP-EGFP fusion gene transgenic mice are established, which harboring mROP-EGFP gene and expressing GFP in their retina. This is valuable for studying the development of brain and retina, pathogenesis of retina disorder and retina transplanting.
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Objective:To evaluate the biological characters of C57-TgN(HBV adr2.0)SMMU transgenic mice. Methods: Integration,expression,replication and histology change of hepatitis B virus gene in F6 transgenic mice were estimated by ge-nomic DNA PCR,Western blotting,ELISA,immunohistochemistry,serum DNA PCR,transmission electron microscopy and H-E staining. Results: Hepatitis B virus gene was integrated into F6 C57-TgN(HBV adr2. 0)SMMU transgenic mice and expressed HBsAg,HBcAg and X protein in liver tissue. HBsAg and HBeAg were expressed in serum of 19. 54% and 3. 39% F6 transgenic mice. Hepatitis B virus were replicated in serum and liver tissue of transgenic mice. Long-term integration,expression and replication of hepatitis B virus gene induced pathological lesion of transgenic mice liver and lung. Conclusion: C57-TgNCHBV adr2. 0)SMMU transgenic mice line has the biological characters including integration of hepatitis B virus gene into genomic DNA,expression and replication of hepatitis B virus gene in serum and liver, and histological change in liver and lung. It is a valuable animal system to study pathogenesis, treatment and prevention of hepatitis B virus.
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The tumor stem cell has attracted more and more attentions in recent years,which is involved in a possibili- ty that the knowledge on the tumor stem cell would be a novel base for the developing of research fields in the tumor pre- vent,diagnose and therapy.However,the basic knowledge system on tumor stem cell has not formed as an enough re- search about it has not been done.In this commentary,the author presents his understanding on several biological issues of tumor stem cells,and some controversy in this field as well.The author also wishes his necessary information would in- duce reader's interesting and future considerations.
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Objective To investigate the effects of patient-controlled intravenous analgesia (PCIA) with lornoxicam and fentanyl on arrhythmia and the expression of platelet membrane glycoproteins in patients with coronary artery disease (CAD) after abdominal surgery.Methods Eighty ASA II or III patients with CAD aged 51-66 yrs weighing 59-68 kg presenting for bdominal surgery participated in this study. CAD was diagnosed by clinical symptoms and ischemic changes on ECG. The patients were premedicated with intramuscular henobarbital 0.1 g and scopolamine 0.3 mg. Anesthesia was induced with fentanyl, droperidol, propofol and vecuronium and maintained with propofol, fentanyl and vecuronium. The patients received PCIA after operation. The PCIA solution contained fentanyl 0.9 mg and droperidol 5 mg in 100 ml of normal saline (N.S. ) in group A ( n - 40) or lomoxicam 56 mg, fentanyl 0.2 mg and dropendol 5 mg in 100 ml N.S. in group B ( n = 40). In group A the loading dose was fentanyl 0.05 mg and in group B lornoxicam 4 mg. PCIA included a background infusion at 2 ml ? h-1 and a bolus of 0.5 ml with a 15 min lock-out. VAS (0 = no pain, 10 = worst pain) was used to measure pain intensity In addition to BP, HR and SpO2 monitoring ECG was continuously monitored with a Holler monitor after operation Blood samples were taken from peripheral vein before and 6h after operation and on the 1st, 2nd, 7th and 8th postoperative days for determination of the expression of CD62p, CD63 and CD41/CD61 on the platelet membrane, platelet count, prothrombin time ( PT) thrombin time (TT) and partial thromboplastin time (PTT) Results The two groups were comparable with respect to sex, age, body weight, severity of CAD, duration of operation and intraoperative blood loss. The patients received no blood transfusion during operation. There was no significant difference in VAS score, platelet count, PT, TT and PIT between the two groups. The incidence of atrial and ventricular premature beat on ECG and the expression of CD41 /CD61 , CD62P and CD63 on the platelet membrane were significandy lower in group B than in group A on the 7th and 8th postoperative days (P
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Genetically engineered mice is one of the powerful research systems in exploring the gene protein life phenomenon,which is useful in generating the laboratory animal model of human diseases and developing the new drugs. With the development of functional genomics and model organism in life science area, biological medicine research is in need of genetically engineered mice deadly. There are still some problems to be resolved, including death effect, no or low expression and no phenotype. It is essential to improve the basic research related to genetically engineered mice.