AIM:To explore the roles of Akt ( also called protein kinase B ) and active caspase-3 in the leptin-mediated chronic morphine antinociceptive tolerance in rats .METHODS: A model of chronic morphine antinociceptive tolerance was established in the SD rats .The protein levels of spinal Akt and cleaved caspase-3 were tested by Western blotting.The technique of immunohistochemical staining was used to detect the immunoreactivity positive cells of phospho -rylated ( p)-Akt and cleaved caspase-3 in the spinal cord .Double staining of immunohistochemistry was used to examine the cellular location of the p-Akt and cleaved caspase-3 positive cells.RESULTS: The chronic intrathecal injection of morphine (15 μg) for 7 d markedly upregulated the spinal protein levels of p-Akt and cleaved caspase-3 in the rats.Thirty min before injection of morphine , intrathecal injection of leptin antagonist (3μg) for 7 d significantly attenuated the upreg-ulation of the protein levels of p-Akt and cleaved caspase-3 induced by chronic morphine treatment .The p-Akt was exclu-sively observed in the spinal neurons .The cleaved caspase-3 was only localized with the spinal astrocytes .Intrathecal injec-ting the inhibitors of leptin , Akt and caspase-3 ameliorated the chronic antinociceptive tolerance .CONCLUSION: The spinal Akt pathway and active caspase-3 are involved in the leptin-mediated chronic morphine antinociceptive tolerance in rats.

Objective:To determine the expression profile and potential roles of CD24 in oral squamous cell carcinoma and explore the values of CD24 function as a potential target of clinical therapy.Me-thods:Semi-quantitative immunohistochemistry was used to construct the expression profile of CD24 in 78 human oral tissues and 59 Hamster buccal pouch tissues.Real-time RT-PCR and Western blot were used to analyze the CD24 expression levels in oral DOK4 cells,oral cancer CAL-27 and WSU-HN6 cells. Then these two cancer cell lines were selected to evaluate the effect of all-trans retinoic acid (ATRA)and CD24 antibody on CD24 expression,and the proliferation and tumorsphere formation capacity of these two cell lines.Results:CD24 expression was found significantly elevated in both human and animal tissues compared with normal and benign tissues (P<0.05),as well as in oral cancer CAL-27 and WSU-HN6 cells compared with DOK cells (P<0.05).CAL-27 and WSU-HN6 cells possess increased proliferative and specific tumorsphere formation capability compared with DOK cells (P<0.05 ).Both ATRA and CD24 antibody were able to effectively inhibit the proliferation and tumorsphere formation of CAL-27 and WSU-HN6 cells (P<0.05).Among them ATRA at least involved partially in the proliferation by down-regulating the CD24 expression (P<0.05 ),while CD24 antibody blocking had no effect on the CD24 expression.Conclusion:CD24 was upregulated in oral cancer and functioned as a potential factor that promoted the proliferation and tumorsphere formation of CAL-27 and WSU-HN6 cells.Both ATRA and CD24 antibody might effectively inhibit the proliferation and tumorsphere formation of CAL-27 and WSU-HN6 cells and function as a potential therapy target.

AIM:To investigate the roles of ATP-sensitive potassium ( KATP ) channels in high glucose-induced cardiac injury and the inhibitory effect of hydrogen sulfide ( H2 S) on the cardiomyocyte injury.METHODS:The expres-sion level of KATP channel protein was tested by Western blot.The cell viability was measured by CCK-8 assay.The number of apoptotic cells was observed by Hoechst 33258 nuclear staining.Mitochondrial membrane potential ( MMP) was exam-ined by JC-1 staining.RESULTS:After the H9c2 cells were treated with 35 mmol/L glucose ( high glucose, HG) for 1~24 h, the protein level of KATP channel was significantly reduced at 6 h, 9 h, 12 h and 24 h, reaching the minimum level at 12 h and 24 h.Pretreatment of the cells with 400μmol/L NaHS ( a donor of H2 S) prior to exposure to HG for 12 h con-siderably blocked the down-regulation of KATP channels induced by HG.Pretreatment of the cells with 100 μmol/L mito-chondrial KATP channel opener diazoxide, 50μmol/L non-selective KATP channel opener pinacidil or NaHS obviously inhibi-ted HG-induced injuries, leading to an increase in the cell viability, and decreases in the number of apoptotic cells and the MMP loss.Pretreatment with 100μmol/L mitochondrial KATP channel antagonist 5-hydroxydecanoic acid or 1 mmol/L non-selective KATP channel antagonist glibenclamide attenuated the above cardioprotective effects of NaHS.CONCLUSION:KATP channels mediate the inhibitory effect of H2 S on HG-induced cardiac injury.

Objective To explore the clinical value of using pectoralis major myocutaneous flap in surgery with various adverse factors for advanced oropharyngeal cancer.Methods 15 patients who presented with advanced oropharyngeal cancer were performed for radical surgery,and pectoralis major myocutaneous flaps were applied for immediate reconstruction of surgical defect.In all cases,various adverse factors(advanced age,diabetes,cardiovascular disease,radiation damage,etc.)were existed.The effect of reconstruction was observed.Results All surgeries were successful,and all patients tided over perioperative period.The average time of operation was 5 hours with a range of 4 to 6 hours.The average time cost for flap preparation was 40 minutes with a range of 40 to 55 minutes.The completely survival rate of flaps was 93%.Partial skin necrosis occurred in 1 case(7%);and submandibular fistula occurred in another one(7%).Two patients(13%)developed wound infection.All of them were healed by symptom-atic treatment.Conclusion For the patients with advanced oropharyngeal cancer and various adverse factors of sur-gery,pectoralis major myocutaneous flap can be used as salvage measure in reconstruction of surgical defect,which is more secure than free flap.

AIM:Tostudywhe ther theangiotens in-(1-7)[Ang-(1-7)]/Mas receptor axis protects cardio-myocytes against high glucose (HG)-induced injury by inhibiting nuclear factor-κB (NF-κB) pathway.METHODS:The cell viability was measured by CCK-8 assay.The intracellular levels of reactive oxygen species ( ROS) were detected by DCFH-DA staining .The number of apoptotic cells was tested by Hoechst 33258 nuclear staining .Mitochondrial membrane potential ( MMP) was examined by JC-1 staining.The levels of NF-κB p65 subunit and cleaved caspase-3 protein were de-termined by Western blotting.RESULTS: Treatment of H9c2 cardiac cells with 35 mmol/L glucose (HG) for 30, 60, 90, 120 and 150 min significantly enhanced the levels of phosphorated ( p) NF-κB p65, peaking at 60 min.Co-treatment of the cells with 1 μmol/L Ang-(1-7) and HG for 60 min attenuated the up-regulation of p-NF-κB p65 induced by HG. Co-treatment of the cells with Ang-(1-7) at concentrations of 0.1~30μmol/L and HG for 24 h inhibited HG-induced cy-totoxicity, evidenced by an increase in cell viability .On the other hand, 1 μmol/L Ang-(1-7) ameliorated HG-induced apoptosis, oxidative stress and mitochondrial damage , indicated by decreases in the number of apoptotic cells , cleaved caspase-3 level, ROS generation and MMP loss .However, the above cardioprotective effects of Ang-(1-7) were markedly blocked by A-779, an antagonist of Ang-(1-7) receptor (Mas receptor).Similarly, co-treatment of H9c2 cardiac cells with 100 μmol/L PDTC ( an inhibitor of NF-κB) and HG for 24 h also obviously reduced the above injuries induced by HG.CONCLUSION:Ang-(1-7)/Mas receptor axis prevents the cardiomyocytes from the HG-induced injury by inhibiting NF-κB pathway .

Objective To investigate the geometric feature of femoral medullary cavity through CT scanning and it's matching with femoral intramedullary nail,and to analysis the reason for difficulties in implanting distal lock pin.Methods Thirty dried femur specimens were measured through CT scanning,and twenty images were taken evenly according to the total length of shaft of femur from each femur.In each image,the data of the central axis point and eight points around medullary cavity wall in X and Y axes were obtained,which were used to reconstruct the three-dimensional models of medullary cavity,central axis,intramedullary nail by computer software.And the models were overlapped for comparison.Then the coronal and sagittal fold line charts for central axis of medullary cavity were drawn.The inserting process of intramedullary nail was simulated to observe whether the nail would punch out of the medullary cavity wall.Results The central axis of femoral medullary cavity and intramedullary nail were overlapped and compared.They matched well in the coronal plane,but the curves of femoral medullary cavity were larger than those of intramedullary nails in sagittal plane.While simulating the inserting process of intramedullary nail,6 nails punched out of the medullary cavity wall in coronal plane (20％,6/30),so did 13 nails in sagittal plane (43％,13/30).Conclusion Intramedullary nails match well with most of the femoral medullary cavities.However,the anatomic structure of the femoral medullary cavity differs individually.Curves of some femoral medullary cavity are large,which can cause deformation of intramedullary nail,and this is the main reason for the failure of distal locking.

Objective To explore whether the phosphorylation of NF-κB P65 subunit is involved in the cytotoxicity to and inflammation in an immortal human keratinocyte cell line HaCaT during cobalt chloride (CoCl2-induced chemical hypoxia. Methods HaCaT cells were treated with CoCl2 of 2 mmol/L to set up a chemical hypoxia-induced cell model of injury. Then, RNA interference was used to down-regulate the expression of P65 in CoCl2-induced HaCaT cells. After additional culture, cell viability was tested by cell counting kit8 (CCK-8), the levels of interleukin 6 (IL-6) and interleukin 8 (IL-8) were detected by ELISA kits, phosphorylated and total P65 protein was measured by Western blot. Results The exposure of HaCaT cells to 2 mmol/L CoCl2 for 0 to 4 hours enhanced the phosphorylation of P65, which began at 0.5 hour, peaked at 1.5 hours, and restored to the normal level at 4 hours, and the level of P65 phosphorylation was about 6.6 times that in the untreated control group. The CoCl2 of 2 mmol/L decreased the cell viability of HaCaT cells in a time dependent manner, and a significant difference was observed in the viability of HaCaT cells between CoCl2-treated and untreated HaCaT cells at 2, 4, and 6 hours (P ＜ 0.05, 0.01, 0.01 ). The release of IL-6 and IL-8 from HaCaT cells was also promoted by CoCl2 treatment. The knockdown of P65 expression with siRNA markedly suppressed the CoCl2-induced cytotoxicity to and increase in the release of IL-6 and IL-8 from HaCaT cells,despite of an increment in cell viability by about 11%. Conclusion The phosphorylated P65 subunit mediates CoCl2-induced cytotoxicity and inflammatory injury to HaCaT cells.

Aim To explore the effect of survivin in PC12 cells against injuries induced by cobalt chloride(CoCl_2).Methods PC12 cells were exposed to CoCl_2 at different doses in different time to set up the chemical hypoxia induced PC12 cells injuries model.Cell viability was tested by using cell counter kit-8.Dose-effect(200～1 000 μmol·L~(-1))and time-effect(0～48 h)relationship between hypoxia induced by CoCl_2 and the expression of survivin was evaluated by western blot.Results PC12 cells viability was inhibited significantly by CoCl_2 in a dose and time dependent manners;At the concentrations from 200 to 600 μmol·L~(-1) CoCl_2 for 24 h,survivin expression was upregulated in a dose dependent manner,peaking at 600 μmol·L~(-1) CoCl_2 treatment,exceeding this concentration of CoCl_2,with dose increasing,survivin expression decreased.At the dose of CoCl_2 up to 1 000 μmol·L~(-1),survivin did not express basically;Treatment with 600 μmol·L~(-1) CoCl_2 in different time,within the range of 0～36 h,the expression of survivin enhanced in time dependent manner.But with the extension of time,survivin expression was declining; 17-allylamino-17-demethoxygeldanamycin (2 μmol·L~(-1)),an inhibitor of Hsp90,not only decreased survivin overexpression but also obviously enhanced the injuries of PC12 cells induced by CoCl_2,which didn't damage PC12 cells alone.Conclusion Upregulation of survivin expression may be one of the endogenous defense mechanisms for PC12 cells against chemical hypoxia.

Objective To investigate the effect of meridian point cosmetic therapy of traditional Chinese medicine(TCM)on acne.Methods 80 cases of acne patients were selected and divided into the treatment group(50 cases)and the control group(30 cases).The treatment group was given medication by differentiation of symptoms and signs and designed a complete treatment scheme by the demonstration of health analysis curve through test by balanced cosmetic equipment and meridian point of TCM.The control group only received extravenous medication.The effect of the two groups was observed after 2 months' treatment.Results The total effeetive rate in the treatment group was 94.0%,which was superior to that of the control group(73.3%),x2=11.08,P＜0.05. Conclusion Application of meridian point cosmetic therapy of TCM could facilitated the diagnosis and treatment by differentiation of symptoms and signs and thus improve the curing rate.

BACKGROUND: Improper practice during military training is likely to cause various training wounds, among which patellar tendinosis is the common one.OBJECTIVE: To explore the onset characteristics of patellar tendinosis caused by military training and incidence changes after the implementation of interventions.DESIGN:Sampling investigation.SETTING: Department of Orthopedics, Guangzhou General Hospital of Guangzhou Military Area Command of Chinese PLA; Department of Surgery, Hospital of Chinese PLA Garrison in Hong Kong PARTICIPANTS: Male army soldiers aged 18-24 years were recruited from a full-time training army in August 2000 (non-intervention group) and August 2001 (intervention group). The same training program was carried out among the 2,783 soldiers in non-intervention group and 5,824 soldiers in the intervention group.METHODS: The investigation group was composed of medical workers with senior and intermediate professional titles. Uniform diagnostic standard was made before the investigation, and questionnaire survey wascombined with on-the-spot inspection on soldiers who complained about knee joint pain following training. Those who conformed to the diagnosis were inquired of their training state in detail and possible causes; meanwhile knee X-ray examination was also conducted. Soldiers in the non- intervention group were subjected to the investigation of the incidence and cause of patellar tendinosis due to fulltime training without given any preventive intervention. By contrast, soldiers in the intervention group were given preventive and therapeutic interventions and then subjected to the investigation into the interventional outcomes one year later.MAIN OUTCOME MEASURES: The incidence of patellar tendinosis in soldiers of the two groups.RESULTS: The first and second investigations were conducted on the 2 783 soldiers and 5 824 soldiers, respectively. All of them entered the rediers of the non-intervention group (the incidence of 0.61%) as compared to 15 soldiers in the intervention group (the incidence of 0.26%) (P＜0.01).tenderness. Patel1ar bone X-ray inspection on 12 of them displayed patellar ciated with run-jump training projects; 23 cases were caused by 400 mbarrier training and 7 cases by 5 km cross-country training.CONCLUSION: Patellar tendinosis during military training is mostly caused by run-jump training and can be remarkably prevented by preventive interventions.