AIM: To study p42/p44 mitogen - activated protein kinases ( MAPK ) signal transduction pathway effect on vascular endothelial growth factor ( VEGF ) expression induced by elevated glucose concentration in cultured human retinal pigment epithelium ( hRPE) .
METHODS:hRPE cells were cultured and divided into four groups:normal glucose group (NG) (5. 6mmol/L), high glucose group ( HG1:15mmol/L D-glucose, HG2:20mmol/L D - glucose, HG3:30mmol/L D - glucose ), PD98059 group: hRPE cells were treated by an efficient and selective inhibitor PD98059 (20μmol/L) of p42/p44MAPK signal transduction pathway and solvent dimethyl sulfoxide group ( DMSO group) . The expression of VEGF and pigment epithelium derived factor ( PEDF ) mRNA was detected by RT-PCR. VEGF protein expression in cultured hRPE supernatants was detected by enzyme-linked immumosorbent assay ( ELISA) .
RUSULTS: VEGF mRNA and protein expression induced by elevated glucose concentration increased significantly. VEGF mRNA and protein expression were restrained in PD98059 group. Ratio of ( VEGF/β-actine)/( PEDF/β - actine ) in PD98059 group decreased significantly compare with that in high glucose group.
CONCLUSION: p42/p44MAPK signal transduction pathway might play a part in VEGF expression induced by elevated glucose concentration in cultured hRPE cells.

The diversity in Tibetan kefir grains and dynamics of the microbial community during the fermentation of Tibetan kefir by PCR-DGGE fingerprinting technique were studied.The results showed that bacterial community of Tibetan kefir grains was more complex than that of yeast.The bacteria communities between different Tibetan kefir grains showed 78%～84% similarity, and yeast 80%～92%.Bands B, E and N of DGGE profiles of the microbial community during fermentation of Tibetan kefir were present throughout the fermentation.Analysis of sequence date showed the majority of the DGGE bands of bacteria community corresponded to LAB, and the most intense band (band E) was completely homology to Lactococcus lactis.

Objective To construct an expression system to produce the virus-like particles containing a part of the sequence of PSA mRNA, which are ribonuclease-resistant due to the encapsulation of the mRNA by bacteriophage MS2 coat proteins. Methods The PCR products of PSA cDNA fragments were cloned to TA vector pBS-T, then the targeted segments could be obtained when the pBS-T-PSA were digested by restriction endonuclease Hind Ⅲ and cloned to prokaryocytic expression vector pNCCL1. The recombinant plasmids named PNCCL1-PSA were transfected into E. Coli BL21-DE3 and induced to express with IPTG. Results The recombinant plasmids were successfully constructed. The bacteriophage MS2 coat protein which expressed in BL21 can self- assemble to form ribonuclease resistant virus-like particles and the PSA mRNA was encapsulated into virus-like particles. Conclusions The virus-like particle containing PSA mRNA can be expressed in prokaryocyte and it can be used as standard and control in detecting PSA mRNA. It provides a new, stable and ribonuclease-resistant RNA standard in RNA detection.

Objective:To evaluate the clinical application of 64-slice spiral computer tomography pulmonary angiography (CTPA)in diagnosis of pulmonary embolism(PE).Methods:Sixty-two patients suspected of PE were examined by 64-slice spiral CTPA.The image findings combined with their clinical data were retrospectively analyzed.Results:Twenty-four of the 62 patients were confirmed to have PE by clinical data,laboratory examination and follow-up examination.64-slice spiral CTPA discovered 152 involved branches in the 24 PE patients,including 4 branches in left and right pulmonary trunk,52 in lobar pulmonary arteries,82 in segmental pulmonary arteries,and 14 in subsegmental arteries.Four types of PE were detected in our group,including eccentric filling defect in 58 branches,central filling defect in 49 branches,total occlusion of the pulmonary arteries in 21 branches,and mural embolism of host artery in 24 branches.The diagnosis accuracy of 64-slice spiral CTPA in the present group of patients was 100%,with no missed diagnosis and misdiagnosis.Besides,64-slice spiral CTPA could reflect the location,morphology,involvement and degrees of PE.Conclusion:64-slice spiral CTPA is a rapid,accurate and non-invasive diagnostic approach for PE.It is the first choice in clinical screening of PE and may serve as a gold standard for diagnosis of pulmonary embolism.

OBJECTIVETo explore mid-term follow up results of distal fixation prosthesis in treating unstable intertrochanteric fractures in elderly patients.

METHODSFrom May 2008 to March 2014,58 elderly patients with unstable intertrochanteric were treated with distal fixation prosthesis, among them, there were 15 males and 43 females aged from 75 to 87 years old with an average of 83.2 years old. Fracture were classified according to Evans classification, 39 cases were type I c and 19 cases were type I d. Surgical risk was evaluated before operation, 9 patients were performed total hip arthroplasty and 49 patients were performed prosthetic replacement hip joint function of patients with different age period, Evans classificaton, prothesis type, fixation method were evaluated respectively by using Harris score.

RESULTSFifty-six patients were followed up from 13 to 36 months with an average of 21.6 months. Harris score was 83.51 ± 6.40, 5 cases got excellent results, 38 cases good and 13 cases moderate. Harris score of patients aged from 75 to 80 years old was 88.64 ± 2.35, 81.64 ± 6.40 in patients aged more than 80 years old, and had significant differences between two groups; Harris score in patients with type Evans I c was 83.64 ± 6.53, and 83.11 ± 6.08 in type Evans I d, while there was no significant differences between two groups. There was no obvious meaning in Harris score between patients with tension band (83.63 ± 6.15) and without tension band (82.41 ± 6.57). There was no significant meaning in Harris score between patients with normal distal fixation prosthesis (83.34 ± 6.43) and femoral moment reconstruction distal fixation prosthesis (83.92 ± 6.51). There was 1 patient occurred hip joint dislocation on the operative side and re-dislocation after manual reduction, then received open reduction. Two patients occurred femoral osteolysis without clinical symptoms, and treated conservative treatment.

CONCLUSIONArtificial joint replacement for unstable intertrochanteric fractures in elderly patients, hip joint function in patients aged more than 80 years old is worse, while there was no obvious market effect in fracture classification, whether to use tension band and type of distal fixation prosthesis, moreover, proximal femoral osteolysis should be focused on.

Aged ; Aged, 80 and over ; Arthroplasty, Replacement, Hip ; Female ; Femur ; surgery ; Follow-Up Studies ; Fracture Fixation, Intramedullary ; Hip Fractures ; surgery ; Hip Joint ; surgery ; Hip Prosthesis ; Humans ; Male ; Treatment Outcome

The role of general practitioners in the UK was analyzed and we found that in the UK, there was normalized general medical education system, perfect general practitioner training sys-tem, specific responsibilities and assessment index, close liaison group between primary and secondary care, strong economic supporting measures and high degree of recognition. Therefore, to deepen med-ical reform and improve basic medical service, we must accelerate general practitioners training, im-prove practice ability of general practitioners, establish the first-treatment system in community, reit-erate role of gatekeeper, enhance the payment of general practitioner and increase their sense of iden-tity. At the same time, we must strengthen the informatization construction, set up the first-treatment in the community and the patency of two-way referral system.

OBJECTIVETo study the effect of excessive fluoride on calcium overload and apoptosis in cultured rat ameloblasts in vitro.

METHODSLogarithmic-phase ameloblasts (HAT-7) were treated with 0, 0.4, 0.8, 1.6, 3.2, and 6.4 mmol · L(-1) sodium fluoride (NaF) solution. Cell activities were detected by using a Cell Counting Kit 8 (CCK-8) assay after 48 h of treatment. The effect of fluoride on cell apoptosis was analyzed by using flow cytometry. Excessive fluoride-induced calcium concentration and calreticulin expression changes in ameloblasts were detected by using laser scanning confocal microscopy, Western blot analysis, and real-time quantitative polymerase chain reaction.

RESULTSNaF inhibited ameloblast activity at 1.6, 3.2, and 6.4 mmol · L(-1) (dose-dependent) after 48 h of induction. The Ca2+ fluorescence intensity of HAT-7 cells incubated with 1.6 and 3.2 mmol · L(-1) NaF was higher than that in the control group. The fluoride-induced early-stage apoptosis of ameloblasts after 48 h of induction and the early-stage apoptosis rate was positively correlated with fluoride concentration. Calreticulin mRNA expression in HAT-7 cells was higher than that in the control group after 48 h of incubation with 0.8, 1.2, and 1.6 mmol · L(-1) NaF.

CONCLUSIONExcessive fluoride-induced calcium overload in ameloblasts and further caused endoplasmic reticulum stress-mediated apoptosis.

Ameloblasts ; Animals ; Apoptosis ; Calcium ; Calcium Fluoride ; Fluorides ; Phosphates ; Rats ; Sodium Fluoride

Objective To review the safety and efficacy of upper-pole access percutaneous nephrolithotomy (PCNL). Methods From May 2008 to May 2010,237 cases ( 135 males and 102 females with mean age of 42 yrs) of renal or proximal ureteral calculi treated with upper-pole access PCNL were reviewed.The indications included calculus larger than 1.5 cm or impacted proximal ureteral calculi in 94 cases,calculus≥2 cm or impacted renal pelvic calculi in 26 cases,staghorn or multiple calculi in 68 cases,complex lower calyx calculi in 13 cases,upper calyx calculi not amenable to ESWL or URS in 12 cases,calculi within upper calyx diverticulum in 3 cases,combined UPJ obstruction or upper ureteral stenosis in 8 cases,morbidly obese patients in 3 cases,calculi within horseshoe kidneys in 6 cases,calculi within transplanted kidneys or ureters in 4 cases.Of the 237 cases,175 tracts (73.8％) were above the 12th rib,46were above the 11th rib,12 were below the 12th rib,4 were in the lower abdomen for renal transplant patients. Results The overall stone clearance rate with upper-pole access PCNL monotherapy was 74.3％.Additional punctures were required in 55 cases and combined ESWL in 6 cases.Total stone clearance rate at 3 months after operation was 88.2％.16 patients (6.8％) had a pleural injury.Thoracentesis was required in 8 patients,closed thoracic drainage in 5 patients,conservative treatment in 3 patients.After nephrostomy tube removed,pleural irritation symptom appeared in 12 cases (5.1％) who required symptomatic treatment.No patient had injury to the lung or other viscera.Significant bleeding requiring blood transfusion was olserved in 5 patients,while selective renal arterial embolization was required in 2 cases. Conclusions Upper-pole access offers optimal visibility,convenience for the movement of rigid nephroscope and high stone-free rate.It should be attempted in selected cages of upper ureteral calculi and complex renal calculi.

Objective To evaluate the efficacy and safety of bladder hydrodistention for the treatment of ketamine-associated bladder dysfunction.Methods Six patients were required to withdraw the ketamine and treated with bladder hydrodistention therapy and sodium hyaluronate irrigation,and medicine to pretect liver and kidney was also used.Results The biopsies of 6 cases demonstrated the cystitis through biopsy.Lower urinary tract symptoms such as urgency,thamuria and odynuria were significantly relieved after bladder installation within 30 days.The O'Leary-Sant ICSI scores and the ICPI scores reduced to 3.5 ± 1.6,2.8 ± 1.5 respectively.The functional bladder capacities increased to an anverage of (180 ± 28)ml,.2-3 times of nocturia,Qmax (14.4 ± 4.3) ml/s.All cases were followed up for 4 to 18 months.Symptoms disappeared or were significantly relieved in all patients.Conclusion Contracture of bladder might be the main presentation of ketamine-associated bladder dysfunction.Intravesical hydrodistention therapy and sodium hyaluronate irrigation could be the safe and effective therapy in the treatment of katamine-associated dysfunction.

Objective To study the methods and results of endovascular treatment of posterior inferior cerebellar artery (PICA) aneurysms. Methods Twenty-one patients with PICA aneurysms were treated with endovascular treatment. The locations of aneurysm on PICA were evaluated through the DSA. Eight patients received single coil embolization, 5 received liquid Glue embolization, 2 received coil embolization combined with liquid Glue, 2 received coil embolization assisted with stents, and 4 underwent ocelasion of the parent PICA. Outcome was evaluated with the Glasgow outcome scale (COS). Results There were complete (100%) occlusion in 5 patients, near complete (>90% ) occlusion in 2, and incomplete (85%) occlusion in 1 in single coil embolization. Seven patients with Glue embolization (n = 5 ) or combination with coils ( n = 2 ) exhibited complete ( 100% ) occlusion. There were near complete ( >90% ) occlusion in 2 cases with coil assisted with stents. Complete occlusion of the parent PICA was achieved in 3 patients, and near complete occlusion of PICA in one case. One patient suffered from new neurological deficits, and one patient treated with coils and stents died. None of the patients suffered from re-bleeding. These patients received follow-up during a mean period of (22±8) months. Overall long-term outcome was good ( GOS score 4 or 5 ) in 17 patients, poor ( COS score 2 or 3) in 3, and fatal ( GOS score 1 ) in one case. Conclusions According to the location of aneurysms on PICA, aneurysms can be effectively and safely treated with endovaecular embolization.