Objective For the purpose of exploring the influence exerued by the amount of template DNA on the results of automatic fluerescent detection. Method For the purpose of buiding the optimal amount of template DNA amplified, we have analyzed different amount of template DNA (9947A) by 3100 Genetic Analyzer with the multiplexing Profiler Plus Kit. Results The optimal amounts of template DNA amplified were between 0.31ng and 2. 5ng by using 3100 Genetic Analyzer. Conclusion It will produce the incorrect analytic results if the amounts of template DNA were too high or too low.

Objective To explore the clinical characters,therapy and prognoses of hyperthyroid cardiac disease in children.Method Fourteen hospitalized cases with hyperthyroid cardiac disease(5-14 years old) collected from January 2001 to January 2005 were analyzed(retrospecti)vely.Results Fourteen children with hyperthyroid cardiac disease had different degree of enlarged of heart,arrhythmia and heart failure.All of the cases were received treatment of hyperthyroidism and anti-arrhythmia.After admission for 2 weeks,the symptoms of 13 cases relived or disappeared,and after 4-6 weeks the enlarged heart,arrhythmia and heart failure disappeared.One case died after recurrent heart failure.Conclusions Hyperthyroid cardiac disease in children can be curative and the changes of cardiac blood-vessel can be recuperative.It is critical for doctors to know the manifestations of hyperthyroidism and take the cardiac examination and the function of the thyroidism and take proper treatment as soon as possible.

BACKGROUND: Peripheral facial paralysis is treated at present only with traditional traumatic methods, such as acupuncture, moxibustion,needling with physical therapy, point-injection, thread burial therapy,scraping therapy, etc. Those methods do not act directly on the lesion of facial nerve and they are long in duration of treatment and low in curative rate. The constant magnetic aerobic atruamatic method along meridian just makes up the shortcoming of the traditional therapies.OBJECTIVE: To observe the therapeutic effects of constant magnetic aerobic atruamatic method along meridian on treatment of peripheral facial paralysis.DESIGN: Case analysis.SETTING: Department of Facial Paralysis, College of Stomatology, Fourth Military Medical University of Chinese PLA.PARTICIPANTS: Totally 1 500 cases of first diagnosed peripheral facial paralysis were employrd in 18 provinces and autonomic regions in China from June 1998 to June 2004, of which, 935 cases were male and 565 cases female. Of 103 cases were aged varied from 1.5 to 19 years, 1 105 cases from 20 to 50 years and 292 cases from 51 to 74 years. Of 672 cases were left palsy and 828 cases were right palsy. The duration of sickness was varied from 1 day to 40 years.METHODS: Constant magnetic aerobic therapy for 20 minutes was applied on facial-palsy effective points for all of cases [No.1 point: in the depression inferior to mastoid process behind ear lobe on the affected side,corresponding to the mastoid foramen, from which, the facial nerve emerges. No.2 point: on the arch top of mastoid process behind ear lobe on the affected side. No.3 point: about 3.33 cm (1 cun) above No.1 point. No.4: in the conjunction between the point about 0.5 cm posterior to the external auricular tract and the one 0.5 cm inferior to helix cms. No.5 point:about 1 cun anterior to ear apex. No.6 point: on the top of the upper border of helix on the affected side] and supplementary points [Qianzheng (Extra),Dicong (ST 4), bilateral Hegu (LI 4)], once per day, 10 days makes one course. There were a couple of days at interval between two courses, totally two courses were required. Point and oxygen therapies were applied simultaneously. Oxygen therapy: 20 minute/time, once per day, 10 days made one course, at the volume of oxygen 2-4 L/minute. Evaluation of therapeutic effects: It was to compare the changes between the affected side and healthy side in forehand wrinkles, frowning, closing eyes, whistling, blowing and smiling and the changes between two sides in nasal labial groove and philtrum during showing the teeth and at static state. Comparison of bilateral muscularis function of healthy people and patients were obtained with the six-point and four-line measurement.MAIN OUTCOME MEASURES: Results of evaluation on clinical therapeutic effects.RESULTS: Totally 1 500 cases of peripheral facial paralysis entered result analysis, no dropped out case. After 20 days treatment, 1 385 cases (92.33%) were recovered, 92 cases (6.13%) appeared significant effect, 22cases (1.74%) were improved and 1 case (0.07%) was no effect. The total effective rate was 99.93%.CONCLUSION: Constant magnetic field in combination with oxygen therapy is characterized as no pain, no trauma, no cross infection and no second injury of facial nerve, safe and convenient operation, short duration of treatment and high curative rate. It improves remarkably the symptoms of facial paralysis and recovers facial function.

Objective To investigate the expression of osteopontin in colorectal cancer and hepatic metastatic cancer and its clinical significance.Methods The expression of osteopontin in 76 cases of colorectal cancer tissues,30 para carcinoma normal mucosa,liver metastatic tumor tissues in 16 patients was examined by immunohistochemical SP staining method.Results The expression rates of osteopontin in normal mucosa,colorectal cancer tissues and liver metastatic cancer tissues were 6.7％ (2/30),69.7％ (53/76),75.0％ (12/16) respectively.The expression rates of osteopontinin colorectal cancer and liver metastatic cancer was significantly higher than those in the normal mucosa (respectively x2 =34.273,23.014,all P ＜ 0.001).The expression of osteopontin was related to infiltration depth,lymph node metastasis,and distant metastasis (respectively x2 =14.347,6.577,7.278,5.537,all P ＜ 0.05).There was no significant difference in the sex and age.Kaplan-Meier survival analysis showed that patients with osteopontin positive had poor prognosis compared with osteopontin negative patients (P ＜ 0.001).Conclusions The expression of osteopontin is closely related to the invasion and metastasis of colorectal carcinoma,a hopeful indicator for the prognosis of colorectal cancer.

Objective To investigate the potential role of tumor necrosis factor receptor-associated factor-6(TRAF6)in the rat cerebral ischema-reperfusion injury.Methods 40 healthy adult SD rats were divided into 5 groups(n=8)according to the ran-dom control principle:sham operation group,ischemia group,reperfusion 2 h group(R2 h),reperfusion 12 h group(R12 h)and reperfusion 24 h group(R24 h).The rat cerebral ischemia-reperfusion injury model was constructed.The change of TRAF6 expres-sion was examined by RT-PCR and Western-blot.Then,the immunohistochemistry was adopted to locate the TRAF6 protein.Re-sults Compared with the sham group,the expression of TRAF6 in the ischemia group and the R2 h,R12 h and R 24 h groups was obviously increased,but the difference had no statistical significance (P <0.05).TRAF6 was mainly located in the cytoplasm of neuronal cells.Conclusion Activated TRAF6 is involved in the brain cell death induced by cerebral ischemia-reperfusion.

As a member of tyrosine kinases(TK) family,Epidermal growth factor receptor(EGFR) has an activity of intrinsic protein tyrosine kinase,which plays an essential role in the regulation of signal transduction in the cells.Due to the abnormal expression of EGFR-TK,the certain type cancers may developed and progressed.Based on that,the inhibitors of EGFR-TK could be effective medicines for the treatment of cancer.The EGFR-TK domain was amplified by RT-PCR with RNA of HUVCEs cells as the template and expressed in E.coliBL21(DE3) using plasmid pET30a as vector.The recombinant protein was purified with the affinity chromatography(Ni-NTA),which was identified to have kinase activity catalyzing the substrate phosphorylated with ATP in the enzymatic reaction.Using the recombinant EGFR-TK as target,the screening model for enzymatic inhibitors was constructed.

Objective Investigate the expression difference of miRNA in cerebrospinal fluid (CSF) to explore new biomarkers for glioma diagnosis and evaluate the diagnostic value .Methods The candidate biomarkers in CSF were detected by using FQ-PCR for 20 cases of glioma patients and 20 cases of non-glioma patients(control group) .miRNAs with significant level changes in CSF sam-ples from patients with gliomas (n=20) compared with healthy volunteers (n=20) were screened out by using Mann-Whitney U test and Kruskal-Wallis test ,and the diagnostic values were evaluated by receiver-operating characteristic curves (ROC curves)and area under the curve(AUC) .Results MiR-128 and miR-21 were differentially expressed in CSF samples from patients with gliomas compared with control group .Expression of miR-21 in glioma is significantly higher than that in the control group(P<0 .05) ,while the expression of miR-128 in gliomas was significantly lower than that in control group(P<0 .05) .AUC was 0 .96 when using only miR-21 as the diagnostic biomarker ,and the sensitivity was 90% ,specificity was 95% .The diagnostic sensitivity and specificity were 100% when MiR-128 and miR-21 combined .Conclusion miR-128 and miR-21 are potential markers for gliomas diagnosis in the CSF .

Objective To investigate the changes of molecule markers of pre-thrombotic state,expression of CD62P,concentration of plasma fibrinogen(Fib),prothrombin time(PT),activated partial thromboplastin time(APTT),thrombin time(TT) in the patients with essential hypertension for providing theoretical evidence for diagnosis and therapy of thrombus complication in early clinical stage.Methods Fifty-three patients with hypertension(grade Ⅰ 17 cases,grade Ⅱ 16 cases,grade Ⅲ 20 cases)were selected and twenty-eight people with medical examination in good health were used as control group.The expressions of CD62P and CD61 were measured by flow cytometry.The concentration of Fib and the levels of PT,APTT,TT were measured by Class coagulation method.Results The expression of CD62P and the concentration of plasma Fib in essential hypertension group were higher than those in control groups(P0.05).Conclusion The essential hypertensive patients are in pre-thrombotic state,the activation of platelet and blood clotting activity increase.Molecular markers should be measured in essential hypertensive patients for prevention and treatment of thrombotic disease.

In order to change the situations such as students’passivity and low enthusiasm in the traditional experimental teaching of Microbiology and improve students’comprehensive qualities,exploration and attempt on experimental teaching reforms including teaching contents,teaching modes,laboratory management and evaluation system of examination were done.Teaching practices showed that the normalized ex- periment syllabus based on the correct orientation of experimental teaching of Microbiology,the actualization of teaching mode of basic skill training-integrated experiment-designed experiment and the management of opening laboratory were effective measures in improving experimental teaching of Microbiology.

This paper employed UPLC-Electrospray Ionization /Quadrupole-Time of Flight-Mass /Mass Spectrometry( UPLC-ESI/Q-TOF-MS/MS) to analyze the chemical constituents in the stems of Nelumbo nucifera. The stems of N. nucifera were extracted with 75% methanol, and we applied an Agilent Zorbax SB-Aq column (2.1 mm x 100 mm, 1.8 μm) to UPLC analysis with water methanol-water( containing 0.05% formic acid) in gradient as mobile phase. The eluates were then detected by ESI-Q-TOF-MS/MS. Results indicated that 22 benzylisoquinoline alkaloids were indendified. Among them, one alkaloid may be a new compound and a component was found in the Lotus for the first time. We fully identify the composition of the Lotus stems for the first time, Which could provides theoretical foundation for further study and utilization of the medicinal resources.
Chromatography, High Pressure Liquid ; methods ; Drugs, Chinese Herbal ; chemistry ; Molecular Structure ; Nelumbo ; chemistry ; Plant Stems ; chemistry ; Tandem Mass Spectrometry ; methods