To investigate the inhibitive effect of the co-culture supernatant of Toxoplasma gondii on the proliferation of human colon cancer cell line sw480 and/or its induced apoptosis and necrosis in vitro,the co-culture model of sw480 cells (1x10~6) was established with different number of Toxoplasma gondii (the concentration of tachyzoites was 2×10~6,4×10~6,8×10~6,16×10~6 respectively),and the co-culture supernatants were gathered 72 hours later.The sw480 cells were treated for different hours with different models of the co-culture supernatant.The parasitism and proliferation of Toxoplasma gondii tachyzoites in sw480 cells was observed by Giemsa staining,and the growth inhibition rates of these cells were investigated by the CKK-8 method.Meanwhile,the apoptosis and necrosis of sw480 cells were detected with transmission electron microscopy (TEM) and fluorescence microscopy with Hochest 33258 staining to observe the morphological changes of sw480 cells.Agarose electrophoresis was used to detect the DNA change of sw480 cells,and flow cytometric analysis was used for investigation of rates of apoptosis and necrosis of these cells stained with Annexin-v-FITC/PI.It was found that the parasitism and proliferation of Toxoplasma gondii tachyzoites in sw480 cells could be observed;and as demonstrated by the CKK-8 method,the inhibitive effect of the co-culture supernatant depended upon the treatment time,and a maximum inhibition ratio of 44.55% was detected at 48 hours.The karyopyknosis and karyorrhexis in the treated sw480 cells were observed under the fluorescence microscope;and the cellular necrosis were observed under TEM.In addition,the DNA fragments in the treated cells were revealed by agarose electrophoresis.In the flow cytometric analysis,a maximum prophase apoptosis rate of 11.54% was detected at 48 hours after treatment with the supernatants,while the rate of prophase apoptosis decreased,and the rates of advanced stage/necrosis increased markedly at the treatment time beyond 48 hours.It is concluded that the co-culture supernatant of Toxoplasma gondii and human colon cancer cell sw480 cell line can induce inhibition of proliferation,apoptosis and necrosis of this cell line in vitro.

Objective: To discuss how to improve the level of diagnosis and treatment about the firstbranchial fistula. Method:16 cases with the first branchial fistula were analyzed retrospectively. Result :Of all thepatients underwent surgery, 14 cases were free from disease postoperatively within 0.5 years follow-up, 2patients underwent re-operation because of recurrence. Conclusion:Knowing about the disease fully, selectingproper surgical incision and possessing skilled surgical technique is important to improve the level of diagnosisand treatment about the first branchial fistula.

Objective To explore the relationship of the mean color vessels density (MCVD) and pathologic microvessel density(MVD) in rectal cancers and their relation with T stags and lymph node metastasis.Methods MCVD were caculated preoperationly with transrectal color power angiography(TRCPA).After operation MVD was assessed immunohistochemically using anti-CD105 monoclonal antibody.The relationships within MCVD, MVD and T stages and lymph node metastasis were analysed.Results There were positive correlation between MCVD and MVD (r=0.763, P＜0.01) in rectal cancer.There were significant difference of MCVD and MVD in both the depth of carcinoma invasion and metastasis of lymphatic nodes( P＜0.05).Conclusions MCVD can display features of the blood supply and distribution of pathologic microvessels and reflect the development of rectal cancers.MCVD is a credible index to choose available treatment and to evaluate prognosis before operation of rectal cancer.

Objective To investigate the therapeutic efficacy of surgical treatment of myocardial bridge. Methods Fifteen patients with myocardial bridge had been treated in our hospital from January 1999 to October 2006. Thirteen patients who had typical angina had been treated regularly by medication, but their symptom remission was not satisfactory. One patient suffered from acute myocardial infarction, and another 1 patient complained of palpitation and syncope. All the patients were given surgical treatment. Surgical strategies included minimally invasive off-pump surgery in 9 patients (supraarterial myotomy in 3 patients, coronary artery bypass grafting in 2, and coronary artery bypass grafting with supraarterial myotomy in 4) and on-pump surgery in 6 patients (coronary artery bypass grafting with supraarterial myotomy in 5 patients and supraarterial myotomy in 1). Results All the operations were successfully completed. The operative duration of off-pump and on-pump surgery was 2.8?1.9 h and 3.5?1.7 h, respectively. The extracorporeal circulation time was 59?37 min. No surgery related death or complications occurred. At 3 months after operation, all the patients were free from symptoms, and electrocardiogram returned to normal in 11 patients. During a follow-up for 0.5~7 years (1.9?1.2 years), recurrent palpitation was seen in 1 patient and others reported no angina. Conclusions Surgical treatment of myocardial bridge has good immediate and long-term results.

Objective To evaluate the applicative value of ultrasound elastography in diagnosis of focal lesions in the parotid and submandibular glands.Methods 30 patients(30 lesions) were scanned by real-time ultrasound elastography and were analyzed in elastography evaluated criteria (5 score method).All the results were compared with the pathological types.Results The elastographic grades of most benign nodules were 1 - 3,while most of malignancy were 4 - 5.The elastographic grades of benign and malignant lesions were of significant difference statistically( P ＜0.01 ).If elastographic grade 4 or 5 were diagnosed as malignancies,the sensitivity,specificity and accuracy for diagnosing malignant l(e)sions in the parotid and submandibular glands were 72.73 ％,84.21 ％,80.00 ％,respectively.Conclusions Ultrasound elastography is useful in the differential diagnosis of focal lesions in the parotid and submandibular glands.

Objective To construct the human IL-24 recombined adenovirus(Ad-IL24)and to observe the effect of Ad-IL24 on ex vivo culture of HL-60 cells.Methods pAdTrack-CMV-IL24 was constructed by PCR with pcDNA3.0-IL24 recombined plasmid as a template,enzyme digestion and ligation.The pAdTrack-CMV-IL24 lineared by Pme Ⅰ was co-transformed into BJ5183 with pAdEasy-1.The pAdEasy-1-pTrack-CMV-IL24 recombined adenovirus vector was lineared with Pac Ⅰ and then transfected in to QBI-293A cells.The Ad-IL24 was obtained and used to infect HL-60 ceils,and the effect on HL-60 cells was tested by LSCM,Mrrr,FCM,ELISA and immunohistochemistry staining.Results The pAdEasy-1-pTrack-CMV-IL24 vector was constructed and the Ad-IL24 was successfully obtained.The effect of inhibiting growth of HL-60 cells and inducing cell apoptosis by IL-24 was proved by LSCM,MTT and FCM.IL-24 down-regulated the expression of anti-apoptosis factor Bcl-2 and increased the secretion of IFN-γ,TNF-α of HL-60 cells.Conclusion Ad-IL24 can inhibit the growth of HL-60 cells and induce cell apoptosis through down-regulating expression of anti-apoptosis factor and increasing the secretion of IFN-γ,TNF-α of HL-60 cells.The human Ad-IL24 may provide a basic study for the future target therapy to tumors.

BACKGROUND:Many internal fixations,such as dynamic hip screw,Gamma screw,proximal femoral nail,angle steel plate,as well as locking proximac femoral plate,are utilized in treating intertrochanteric fractures,especially the dynamic hip screw However,the failure rate is gradually increased.OBJECTIVE:To explore the application and clinical efficacy of total hip replacement for the treatment of elderly intertrochanteric fracture fixation after failure of dynamic hip screw.METHODS:A total of four cases with intertrochanteric fractures were treated by total hip replacement after failure of dynamic hip screw fixation was selected.According to Evans typing,one case were type Ⅱ,two cases were type Ⅲ A,and one case was typeⅢB.Internal fixation displacement could be found at half to 1 year after dynamic hip screw fixation.Because of bone disunion,coxa adducta and pain,the patients could not walk.Sequentially,total hip replacement was performed with 45°abduct angle and10°-15°anteversion angle The clinical efficacy was evaluated by Harris scoring criteria.RESULTS AND CONCLUSION:All the cases were operated smoothly,with 1.5-2 hour operation duration and 400-600 mL blood loss.No case appeared allergic response to bone cement.By 3-12 months follow-up,4 successful operative cases do not appear prosthesis loosening,subsidence and are satisfied with well hip function.The average Harris score were 81 points.The results revealed that application of total hip replacement after the failure of DHS in elderly intertrochanteric fracture fixation,which shortened the time patients stay in bed to reduce complications and improved the hip joint function.

Objective To investigate the efficacy of dexmedetomidine on anesthesia analgesia and postopera-tive cognitive dysfunction (POCD)in elderly patients with lumbar surgery.Methods Ninety elderly patients with lumbar surgery were randomly divided into dexmedetomidine group (A group with 46 cases)and control group (B group with 44 cases)by the random number table method.A group received load dosage (1μg/kg)dexmedetomidine before anesthesia induction,and this process must last for more than 10 minutes,then the dexmedetomidine was main-tained at a speed of 0.5μg/(kg·h)during the operation.B group were given the same dosage normal saline in the same way instead.The amount of intraoperative sedation drugs was observed and analyzed in the two groups.MMSE was measured at one day before surgery and seven days after surgery.And,the incidence rate of POCD was compared between the two groups.Results The dosage of intraoperative sedation drugs of fentanyl[(0.57 ±0.11 )mg vs (0.78 ±0.13)mg;t =8.286,P =0.000],propofol[(522.5 ±137.2)mg vs (734.2 ±175.8)mg;t =6.384,P =0.000]and remifentanil[(0.92 ±0.26)mg vs (1.38 ±0.73)mg;t =3.947,P =0.000]in A group were significantly lower than those of group B.After treatment for 7 days,the MMSE score in A group[(27.57 ±1.58)points]was higher than that of B group[(25.02 ±2.14)points](t =6.451,P =0.000).The incidence rate of POCD in A group (6.52%)was significantly lower than that of B group (22.73%),and the difference was statistically significant (χ2 =4.779,P =0.028).Conclusion In elderly patients with lumbar surgery,the dexmedetomidine can cut down the dosage of intraoperative sedation drugs,and it also could reduce the incidence of POCD.

Objective To prepare RGD and TAT co-modified paclitaxel loaded liposome(RGD/TAT-LP-PTX)for A 549 cells targeting.Method The co-modified liposome was prepared by film-ultrasonic method. The appearance,particle size,Zeta potential were evaluated. The cellular uptake by A 549 cells in vitro was used to evaluate the targeting efficiency. The anti-proliferation efficiency of RGD/TAT-LP-PTX was evaluated by MTT assay. Results The particle diameter of the co-modified liposome was (118.5±11.4) nm with the Zeta potential of (21.58±2.42 )mV. The entrapment efficiency of PTX was 86.5%. The result demonstrated that the co-modified liposome uptaken by A 549 were 2.1, 2.8 times higher than that of TAT-LP and RGD-LP, respectively. The RGD/TAT-LP-PTX shows the highest anti-proliferation efficiency. Conclusion The co-modified liposome might serve as a promising tumor delivery system of antitumor drugs.

Objective:To construct the cells which can express LIF protein steadily and study the effect of rhLIF and IL-24 on HL-60 cells.Methods:ECV-304 cells were infected by eukaryotic expression plasmid pcDNA3-LIF,then the positive cells were selected by G418.The positive cells were collected and their culture supernatant was stored for further use.Expression of LIF mRNA was detected by RT-PCR.The pAdeasy-1-pAdTrack-CMV-IL-24 was extracted from DH5a.The recombined adenovirus vector was lineared with PacI and transfected into 293 cells.The IL-24 recombined adenovirus(Ad-IL-24) was obtained and used to infect HL-60 cells.At the same time,the culture supernatant containing rhLIF was added into the HL-60 cells which was identified as positive expression of IL-24 by RT-PCR.Synergistic effect of the cytokines on HL-60 cells was tested by LSCM, FCM and immunohistochemistry stain assay.Results:The cells expressing LIF protein steadily were constructed successfully, the high titer of the recombined adenovirus(Ad-IL-24) was obtained. Expression of IL-24 in infected HL-60 cells was identified by RT-PCR.The apoptosis of HL-60 cells induced by rhLIF and IL-24 was proved by LSCM ,FCM and immunohistochemistry stain assay. They had synergistic effect.Conclusion:rhLIF and IL-24 can inhibit growth of HL-60 cells and induce apoptosis of the cells.They have synergistic effect.