Objective To discuss the effect of problem-based learning(PBL)model in Neurological history collection.Methods PBL model was performed in Neurological class during the period of novitiation,and estimated by the students.Results PBL model has more advantages in improving autonomous study motivation,and brings up accurate clinical learning method.Con-clusions Compared with the traditional model,PBL model was more suitable for the development of modern medical medicine.

Objective To determine the effect of cheA gene of Helicobacter pylori in the bacterial chemotaxis in vitro and colonization in vivo. Methods The entire cheA and cheY genes were amplified and cloned from genomic DNA of H. pylori NCTC11637 strain. Subsequently, the prokaryotic expression systems of cheA and cheY genes were generated and the target recombinant proteins rCheA and rCheY were extracted by Ni-NTA affinity chromatography. Rabbits were immunized with either rCheA or rCheY for obtaining antisera, and rCheA-IgG and rCheY-IgG in the antisera were prepared using ammonium sulfate precipitation plus DEAE-52 column chromatography. A suicide plasmid of cheA gene was constructed and then a cheA gene knock-out mutant ( cheA - ) was generated based on homologous recombinant exchange using the suicide plasmid. The cheA- mutant was identified using PCR and sequencing. The phosphorylation levels of CheA and CheY molecules of cheA - and wild-type strain were determined by using rCheA-IgG and rCheY-IgG anchoring the target proteins and protein phosphorylation detection kit. The differences of chemotaxis in vitro and colonization in vivo between cheA- mutant and wild-type strain were compared using chemotactic model and BALB/c infection model of H. pylori. Results The cheA gene knock-out in genome of cheA- mutant was confirmed by the results of PCR and sequencing. After treated with 0. 001-0. 1 mol/L HCI for 10 min, the phosphorylation levels of CheA and CheY molecules of wild-type strain were rapidly descended from ( 59.6 ±11.5) μmol and (55.5 ± 10.2) μmol to ( 10.8 ± 2.6) and (5. 5 ± 1.2) μmol (P ＜ 0.05 ), while the phosphorylation of CheY molecule of cheA - mutant was no markedly changed with a persistent lower level ( P ＞0.05). The diameters [(10-20) ± (2-3) mm] of chemotactic aggregative rings of cheA- mutant were significantly less than those [(16-24) ± (2-3)mm] of wild-type strain (P ＜0.05). The positive isolation rate (90%) of H. pylori in gastric biopsy specimens of mice that infected with wild-type strain was remarkably higher than that (40%) of mice that infected with cheA- mutant (P ＜0.05). The result of fluorescence quantitative was also showed that the numbers (6.3 × 103 ±2.1 × 103 copies/mg) of H. pylori in gastric biopsy specimens of wild-type strain infected mice were significantly larger than those (8.3 × 101 ±3. 1 × 101 copies/mg) in gastric biopsy specimens ofcheA- mutant infected mice (P＜0.05). Conclusion The cheA gene of H. pylori has an important role in the bacterial chemotaxis in vitro and colonization in vivo.

Objective To evaluate the value of color Doppler flow imaging (CDFI) in the diagnosis of elderly femoral arterial pseudo aneurysm and treating with ultrasound-guided compression after percutaneous transluminal coronary angioplasty (PTCA). Methods Twenty-six elderly patients who were found a mass and/or vascular murmur in the puncture region were examined by CDFI, and the results of CDFI were proven by clinic and MRI. All pseudo aneurysms were repaired by CDFI guidance to compress simply and/or injecting directly batroxobin into the pseudo aneurysm lumen combined with compression.Results Pseudo aneurysms were diagnosed definitely by CDFI in 26 cases,and the accuracy rate was 100%. Twenty-two cases were repaired with CDFI guidance to compress simply and percutaneously injection of batroxobin into the pseudo aneurysm lumen combined with compression occurred in 3 cases. One case was cured by surgery after failing to be repaired with CDFI guidance. Conclusions CDFI has an important diagnosis value of pseudo aneurysm after PTCA. The method of CDFI guidance compression and batroxobin injection for repairing pseudo aneurysm is simple, safe and effective.

BACKGROUND: Up to now, few studies related to the mechanism of low-frequency transcutaneous electrical nerve stimulation (TENS) in relieving pain, and the effect of low-frequency TENS on the activity potential of dorsal horn cells in rats after peripheral nerve injury. OBJECTIVE: To observe the effects of low-frequency TENS on the activity potential of dorsal horn cells induced by mechanical allodynia and thermal allodynia by using animal models of peripheral nerve injury, and observe the efficacy after interfering of naloxone. DESIGN: A randomized control animal study. SETTING: Department of Neurology, Affiliated Hospital of Medical College, Yanbian University. MATERIALS: The experiment was carried out in the central laboratory of Medical College, Yanbian University between February and October 2004. Eighty male Sprague-Dawley rats were used, and 60 random selected ones were operated to separate sciatic nerve, two branch tibial nerves and sural nerves of sciatic nerve were amputated after ligation, and peroneal nerve was left as the experimental group; the other 20 rats were placed at the origin after sciatic nerve was separated, and then the skin was sutured as the control group. METHODS: ① Pain detection (Behavioral test): At 1 week postoperatively, the rats were given mechanical allodynia and thermal allodynia once every 5 seconds for 10 times, and then the frequency of foot withdrawal was detected (0%-40% for mild pain, 40%-70% for moderate pain; 70% and above for severe pain). ② The spontaneous activity potential of dorsal horn cells and that induced by mechanical allodynia and thermal allodynia were detected among the rats with moderate and severe pain in the control group and study group. ③ Low-frequency TENS of 3 mA and 10 Hz was applied to the legs of rats in the experimental group with annular electrode for 10 minute, and the membrane potential of dorsal horn cell was detected before and after stimulation. ④ At the same time of low-frequency TENS was given, rats in the experimental group were injected with naloxone via audal vein, and the membrane potential of dorsal horn cell was detected before and at 10 minutes after naloxone injection.RESULTS: Finally 80 rats were involved in the analysis of results after compensation. ① The foot withdrawal frequencies induced by mechanical allodynia and thermal allodynia in the experimental group were obviously higher than those in the control group (P ＜ 0.01). ② The membrane potential of dorsal horn cell by mechanical allodynia and thermal allodynia in the experimental group were obviously higher than those in the control group (P ＜ 0.01). ③ The membrane potential of dorsal horn cell by mechanical allodynia and thermal allodynia at 10 minutes after low-frequency TENS in the experimental group were obviously higher than those in the control group [(102.6±0.9), (136.9±1.46) impulses per 10 seconds; (175.2±1.28), (240.8±1.51) impulses per 10 seconds, P ＜ 0.01]. ④ In the experimental group, the membrane potential of dorsal horn cell by mechanical allodynia and thermal allodynia at 10 minutes after naloxone injection were obviously higher than those before injection [(174.5±0.4), (235.4±1.4) impulses per 10 seconds, P ＜ 0.01].CONCLUSION: Low-frequency TENS can effectively inhibit the activity potential of spinal dorsal horn cells induced by non-noxious stimulation,and the intravenous injection of naloxone (8 mg/kg) can recover it to the pretreatment level, indicating that low-frequency TENS may alleviate pain by stimulating central nervous system to make it secrete endogenous opium system, and acting on spinal dorsal horn cells to reduce the activity.

Objective To study the value of leptin,adiponeetin and interleukin (IL-1,IL-6,IL-8) in diagnosis,treatment,and prognosis of cerebral infarction.Method The levels of serum leptin ,adiponectin and IL-1,IL-6,IL-8 in acute cerebral infarction before and after treatment were determined with ELISA as well as in the healthy controls.The relationship between these indices and nerve function injury was studied.Results The levels of serum leptin and IL-1,IL-6,IL-8 in the patients increased significantly,but the controls didn't (P

Background Corneal blindness caused by ocular surface disease is one of the main reasons for the global blinding corneal diseases.With the development and progress of tissue engineering technology,tissueengineered cornea offers a new approach to the treatment of ocular surface disease.Objective This study was to obscrve the growth and differentiation of human umbilical cord mesenchymal stem cclls (UC-MSCs) on thc corneal stroma of receipts and investigate the feasibility of human UC-MSCs differentiated into corneal epithelium-like cells and the reparation of injury cornea.Methods Human UC-MSCs were isolated from human umbilical cord using collagenase Ⅳ digestion and passaged in DMEM/F12 containing fetal bovine serum in vitro.The immunophenotype of cultured human UC-MSCs was evaluated by flow cytometry.The differentiated osteoblasts from the human UC-MSCs by directional induce was identified.Twenty-four New Zealand albino rabbits were randomly divided into 2 groups.The human UC-MSCs were cultured on porcine corneal matrix without corneal epithelium for 4 days and then transplanted onto the 12 left eyes of 12 New Zealand albino rabbits,and porcine corneal matrix without corneal epithelium was transplanted onto the left eyes of other 12 New Zealand albino rabbits as control group.The rabbits received keratoplasty were examined using in vivo confocal microscope through focusing(CMTF).The eyeballs were taken off after 2,4 and 8 weeks,the growth and differentiation,expression of cytokeratin 3 (CK3),CK12 and ATP-binding cassette superfamily G memben 2 (ABCG2)of human UC-MSCs were observed by histopathology and immunofluorescence staining.This use of the experimental animals complied with ARVO Statement.Results Digestive human UCMSCs formed round in shape and was large in size.The attached cells displayed long-fusiform shape like fibroblasts.The cultured human UC-MSCs phenotype was CD105+/CD29+/CD44+/CD34-/CD45-and could be induced toward osteoblast differentiation under the appropriate experimental conditions.Human UC-MSCs grew well on the porcine corneal matrix.The corneal grafts survived wcll without rejection till the experiment end in experimental eyes,but the rejection of corneal graft occurred in control eyes.Confocal microscope could observe corneal epithelium-like cells.The corneal epithelium cells showed the positive response for CK3 and CK12 and absent response for ABCG2.Conclusions Human UC-MSCs with porcine corneal matrix can survive,proliferate and differentiate into corneal epithelium-like cells after transplanting onto the corneal stroma of rabbits.This result suggests that human UC-MSCs is able to repair and reconstruct the injured corneal surfaces.

Objective To investigate the clinical features,diagnosis,treatment and prognosis of bladder hemangio-ma. Methods The clinical data of 12 patients with bladder hemangioma were retrospectively reviewed and analyzed in com-bination with relevant literature. Results Ten patients were treated with partial cystectomy,and two patients treated with transurethral resection of bladder tumor (TUR-BT). All patients were diagnosed as the bladder hemangioma by postoperative pathology. Patients were followed up from 4 months to 6 years. There were no recurrence and metastasis in all cases. Conclu-sion Bladder hemangioma is a rare benign tumor, which can be preliminarily diagnosed by combinating with medical imag-ing. The final diagnosis depends on the pathological examination. Treatment options should rely on the factual situations. The partial cystectomy is the first choice for the treatment of bladder hemangioma. The prognosis is good.

Objective To establish a rapid,accurate and specific method to detect the common mycobacteria based on multiplex real-time PCR.Methods The dual priming oligonucleotide ( DPO)primers and TaqMan probes labeled with FAM,ROX,HEX or JOE fluoresceins at 5' end and eclipse at 3' end respectively were designed to detect the 16S rRNA of mycobacteria.Both specificity and sensitivity were estimated on multiplex real-time PCR detecting genome DNA from 4 mycobacterial model species.Sixty eight early morning sputum specimens collected from hospitalized patients in the Red Cross Hospital of Hangzhou were detected by multiplex real-time PCR,bacterial culture and smear microscopy simultaneously.The positive rates were analyzed by chi-square.Results Mycobacteria including Mycobacterium tuberculosis and three common non-tuberculosis mycobacteria spp.were identified by multiplex real-time PCR accurately and specifically,with the limited load at 101 cfu/ml.In 68 sputum specimens,31 were positive (positive rate 45.6％ ) by this method,which was significant higher than that by smear microscopy ( positive rate 14.7％,x2 =15.4,P ＜0.05 ).The positive cases were identified as 28 Mycobacterium tuberculosis,1 Mycobacterium avium and 2 Mycobacterium intracellulare in agreement with the culture results.One case,which is detected by culture,but not by PCR,was identified as Mycobacterium chelonae by sequencing.Conclusion The multiplex real-time PCR characterizing as sensitive,specific and time-saving for Mycobacterium tuberculosis and common non-tuberculosis mycobacteria could be chosen as the rapid laboratory test of mycobacterial infection.

OBJECTIVE: To evaluate the effect of integrated traditional Chinese and Western medicine therapy on the progression of acute renal failure in patients with chronic renal insufficiency. METHODS: Thirty-two patients with chronic renal insufficiency developed acute renal failure recently were treated with Chinese herbs and western drugs intravenously and clysterizing of Chinese herbs liquid for 30 minutes, and the treatment course was 14 days. Assessment of liver and renal function, blood routine, electrolytes and endogenous creatinine clearance rate (Ccr) was performed before and 2 weeks after the treatment. RESULTS: The levels of hemoglobin (HB), white blood cell count (WBC) and serum electrolytes showed no significant changes after the treatment. The levels of blood urea nitrogen (BUN) and serum creatinine (SCr) decreased, while the level of Ccr increased significantly (P<0.05) after the treatment. The total effective rate was 65.6%. CONCLUSION: The integrated traditional Chinese and Western medicine therapy can effectively delay the deterioration of renal function in patients with chronic renal insufficiency accompanied by acute renal failure.

Objective:To assess the feasibility and accuracy of real-time three dimensional echocardiography(RT-3DE) in quantitative evaluation of left ventricular(LV) volume and ejection fraction(EF) in patients with ventricular aneurysm and myocardial infarction.Methods: Twenty-three patients with left ventricular aneurysm combined with myocardial infarction were examined by RT-3DE,two dimensional echocardiography Simpson's method,and M-mode Teichholz method separately.The following parameters: left ventricular end diastolic volume(LVEDV),end systolic volume(ESV),stroke volume(SV) and EF were obtained by each method and the results were compared with those obtained by left ventriculography(LVG).Results: The values of LVEDV,LVESV,SV,and EF determined by RT-3DE showed good correlations with those determined by LVG(r=0.92,0.90,0.88,and 0.91,respectively;P(0.05).) The values of LVEDV,LVESV,SV,and EF determined by Simpson's method also showed good correlations with those determined by LVG(r=0.85,0.87,0.86,0.91,respectively;P