Objective To develop a method for the determination of the dynamic parameters of interaction between methotrexate(MTX)enantiomer and dihydrofolate reductase(DHFR).Methods An affinity capillary electrophoresis(ACE)method was adopted.Using bare fused-silica capillary,the electrophoresis buffer was 50 mmoL/L sodium tetraborate with 0.2%Brij-35,pH 9.50.The temperature of separation was controlled at 25℃ and a voltage of 25 kV was applied.The separation of the reaction mixture was performed at a wavelength of 254 nm.The difference of peak areas about the product was used to calculate the inhibitory rate and IC50 values.Results We establish the detection method for the dynamic parameters of interaction between MTX enantioner and DHFR.The separation of the reaction mixture could be achieved within 30 min.The IC50 value of D-(-)-MTX and L-(+)-MTX were 3.17×10-7 and 2.48×10-8mol/L,respective.The IC50 value of the D-(-)-MTX was 31.67×10-8mol/L,the L-(+)-MTX was2.48×10-8mol/L.The IC50 value of the D-(-)-MTX was about 13 times higher than that of the L-(+)-MTX.Conclusions ACE is a rapid.simple and accurate method that can be used to monitor DHFRdynamic reaction.The IC50 values of MTX enantiomer were quite different.The result first indicated that reaction between MTX enantiomer and DHFR had three-dimensional selection.

OBJECTIVE: To study the mechanism of herbal application along meridians for treatment and prevention of asthma by using serum pharmacological test to observe the effects of serum containing herbs against the constriction of tracheal spiral strips induced by acetylcholine chloride (Ach). METHODS: Guinea pigs were randomly divided into normal control group, normal saline (NS) application group, aminophylline application group, aminophylline injection group, 1-day herb application group, 7-day herb application group and 14-day herb application group. Asthma was induced by Hutson's method in guinea pigs except the normal control group. Guinea pigs in herb application groups were treated by external application of a compound herbal medicine 60 min once every day. Guinea pigs in NS application group were treated by external application of NS. Guinea pigs in the two aminophylline-treated groups were treated by external application and intraperitoneal injection of aminophylline at a dose of 400 mg/kg, respectively. The guinea pigs were killed and the sera were obtained after 1-day, 7-day and 14-day treatment in the three herb application groups, 7-day treatment in the NS application group, the aminophylline application and injection groups, respectively. Serum pharmacological method was used to do the experiment, the effects of different sera on the constriction of tracheal strips were observed, and the constriction rates were calculated. RESULT: The serum containing herbs had an effect in reducing the constriction of tracheal spiral strips induced by Ach, and the effect was similar to that of the serum obtained from the aminophylline injection group. The constriction rate of the tracheal spiral strips was decreased when herbal application treatment was prolonged within a period of time, and it became stable when herbal application treatment was between 7-14 days. The constriction of tracheal spiral strips induced by Ach could be reduced remarkably when it was previously treated by serum containing herbs. CONCLUSION: The anti-acetylcholine function with a time-dependent effect is one of the mechanisms of herbal application treatment along meridians for asthma, and furthermore, herbal application treatment along meridians might be useful for preventing asthma.

Objective To induce mice bone marrow(BM) mono-nuclear cells into dendritic cells(DC) in vitro,and then produce function vaccine of anti-myeloma by bone marrow derived dendritic cells(BM-DC).Methods Mono-nuclear cells separated from mice bone marrow were cultured in tissue culture plastics which supplemented granulocyte-macrophage clonystimulating factor(GM-CSF),IL-4.During the 8 days,there were numerous of mature DCs outgrown,and then fused with SP 2/0 myeloma cells in logarithmic growth phase cultured with 8-azaguaine by polyethylene(PEG).The resulting purified DC/myeloma hybrids were generated by selecting with hypoxanthine-aminopterin-thymidine(HAT).The specific anti-tumor activity was examined in vitro.Results Mono-nuclear cells derived from BM cultured by supplement propriate cytokines milieu in tissue culture plastics,and then exposed to lipopolysaccharide(LPS).There were plenty of mature DCs appear as early as day 8,and exhibit dramatic veils of cytoplasm and extensive dendrites in their surfaces,high express CD11c(83.19%),major histocompatibility complex(MHC)-Ⅱ(95.25%),CD86(89.24%),their capacity to stimulate primary T cell responses in mixed leukocyte reaction(MLR) would be shown strongly.Hybrids could grow well through PEG fusion and HAT selection.These hybrids could stimulate naive T cells into cytotoxic T-lymphocyte(CTLs) directed against SP 2/0 myeloma,and exhibit strong specific killed ability.Conclusions BM-DCs is fused with tumor cells ensure the tumor associated antigens could be processed and presented effectively,and stimulate specific anti-tumor immunity so that prevent or cure this kind of tumor.So it's feasible in practice to produce anti-tumor vaccine by BM-DCs.

Objective To analyze causes of death and risk factors of acute myocardial infarction(AMI)、Methotis 118 AMI patients who died f selected from 1252 hospitalized patients with AMI from January 2003 to June 2008)were retrospectively enrolled for analysis of risk factors and death causes.Resuits The overall mortality of hospitalized patients with AMI was 9.42%(118/1252).The mortality rate in the males was 8.91%(84/943)while in the females was 11.00%(34/309)which was higher than the males but there was no statistical difieFence (P=0.2739).Mortality rate rose along with age and showed significant statistical difference(P＜0.0001)among different age group[＜40 yrs:6.45%(2/31),40～54:2.56%(6/234),55～64:5.11%(16/313),≥65:13.95%(94/674)].Pump failure occurred in 77 cases(65.25%)which was the main cause of death,cardiac arrest occurred in 21 cases(17.80%)and heart rupture in 13 cases(11.02%).There existed other causes of death including cerebral hemorrhage.digestive tract bleeding and pneumonia in 7 cases(5.93%).The mortality of patients with PCI was 4.24%(39/920)while23.80%(79/332)of those witbout PCI(P＜0.0001).Rate of cardiac rupture was 1.04%(13/1252),2.91%(9/309)in females and 0.42%(4/943)in males(P＜0.0001).The time was＜24 h(23.72%.28/118)when death occurred from onset,24 h～1 week(55.93%,66/118)and 1～4 week (20.34%,24/118).There was no statistical difference of mortality related to different infarction locations[antior 12.47%(59/473),anteroseptal 9.23%(12/130),inferior 6.73%(28/416),lateral 8.70%(4/46),ventricle postwall 5.97%(4/67),and ST-segmental elevated myocardial infarction 9.17%(11/120)(P=0.0852)].Conclusions There is a high mortality in aged patients with AMl with heart failure as the most common cause of death which usually occurs at early stage of AMI.The females have more cardiac ruptures than the males.PCI significantly decreases rates of mortality and cardiac rupture.Moreover.gender and location of AMI might be another important risk factor which affect mortality.

Objective: To observe the effect of Yi Jin Jing (Sinew-transforming Qigong Exercises) on the muscle strength in senile sarcopenia. Methods: Sixty-five old people with sarcopenia were randomized into Yi Jin Jing group and a blank control group. Thirty-three patients in Yi Jin Jing group practiced Yi Jin Jing (Sinew-transforming Qigong Exercises), while 32 patients in the blank control group didn't receive any interventions. The muscle strength was measured before and after 12-week training. Results: During the study, each group had 1 dropout. The muscle strength was improved after 12-week training in Yi Jin Jing group, and the difference was statistically significant (P<0.05); there was no significant difference in the blank control group (P>0.05). After the intervention, there was a significant difference between Yi Jin Jing group and the blank control group in comparing the muscle strength (P<0.05). Conclusion: Constant Yi Jin Jing (Sinew-transforming Qigong Exercises) training can notably improve skeletal muscle strength in senile sarcopenia.

Objective To review the experience in diagnosis and treatment of pheochromocytoma in a single center. Methods A total number of 142/145 pheochromocytoma cases treated surgically in our institute from August 2002 to February 2010 were retrospectively reviewed. The mean diameter of tumor was 5.9 cm (1.3- 18. 0 cm). The majority of the tumors (92.9%) were adrenal pheochromocytomas. Ninety-eight patients (69.0 % ) presented initially with hypertension, whereas 44 patients (31%)presented with adrenal incidentaloma. A specific anti-hypertensive pre-surgery preparation with phenoxybenzamine or doxazosine mesylate was started over 1 week before the operation.Of the 142 patients, 91 accepted open surgery, 54 accepted laparoscopic surgery, of which, 5 converted from laparoscopic surgery to open surgery. Results Histopathological results showed that all the cases were pheochromocytoma, while 83 cases were benign, 23 cases were malignant and 37 cases were suspected malignant. Sudden rising of blood pressure during operation was related to the preoperative serum level of catecholamine. Eighty-seven of 98 patients with preoperative hypertension had normal postoperative blood pressure; the remaining 11 patients reduced the dosage of anti-hypertension medication postoperatively. During the follow-up of 3-96 months (median 46 months), 10 of 91patients had a recurrence or metastasis. Six patients died of recurrences or metastasis within 5 years.Conclusions The procedures of qualitative and locative diagnosis of phechromocytoma include clinical manifestations, biochemical tests and imaging investigation. Surgical excision is the fundamental treatment for cure. Patients with high serum level of catecholamine tend to have a sudden rising of blood pressure during operation. Preoperative management is extremely important for the safety of the patient. Intensive follow up is necessary.

Objective To investigate the applied perineal anatomy of the pudendal nerve of female and its direction to perineal operation.Methods The gross anatomy were done on 20 female cadaver(ensuring the pelvic,the hip and the perineum were intact).In each cadaver,the dissection proceeded from intrapelvic,the pudendal canal(Alcock'canal) and the ischiorectal fossa.The composition,track,abut,ramus and domination area of female pudendal nerve were examined.Results The gross anatomy showed that the pudendal nerve was composed of the ventro-branches of sacral nerve 2~4(S_2~S_4).The pudendal nerve sends off anal branch at level of the ischial spine.And the trunk courses in the internal pudendal canal forwardly and inwardly send off the perineal nerves and dorsal nerve of clitoris.The perineal nerves sends off branches to back lunar labium,muscle and urethra.The dorsal nerve of clitoris sends off the nerve to skin of clitoris,and some to the urethra simultaneously.The pudendal nerve has different abut in different anatomical position.Conclusions The perineal anatomy of the pudendal nerve has the meaningful direction to the perineal operations.

Objective To study the clinical efficacy of platelet-rich plasma (PRP) in the treatment of knee osteoarthritis (KOA) and evaluate whether the age,body mass index and grade of KOA are associated with the treatment outcomes.Methods Using the prospective,randomized,controlled study,100 KOA patients hospitalized between December 2013 and November 2014 were enrolled.Twentyeight patients were men and 72 were women.Mean age was 58 years (range,35-85 years).Degenerative arthritis occurred in 68 patients and traumatic arthritis in 32 patients.Kellgren-Lawrence (K-L) score was grade Ⅱ in 35 patients,grade Ⅱ in 46 and grade Ⅲ in 19.The patients were assigned to receive hyaluronic acid (HA) (HA group,n =50) and PRP (PRP group,n =50) by an intraarticular route once weekly for 3 weeks,according to the random number table.Between-group differences were insignificant in age,gender,body mass index (BMI) and K-L grade.Western Ontario and McMaster Universities Arthritis Index (WOMAC),visual analog scale (VAS) and cartilage lesions score (CaLs) were used for clinical and MRI evaluations.At follow-up evaluation,the effective rate was defined at least 36％ improvement from the baseline WOMAC score.Results All patients were followed up for 6 months.The effective rate in PRP group was 84％ versus 68％ in HA group after the last treatment (P ＞0.05),and was 60％ versus 36％ in HA group at the final follow-up (P ＜ 0.05).WOMAC score in both groups had significant improvement after operation,while VAS improved only in PRP group (P ＜ 0.01).In PRP group patients with K-L grade I had better VAS and WOMAC scores than those with grade Ⅱ (P ＜0.05),and patients with grade Ⅱ had better WOAMC score than those with grade Ⅲ (P ＜ 0.05).MRI findings showed seven patients in PRP group had similar CaLs before and after operation (P ＞ 0.05),and the area of abnormal signal in subchondral bone and the depth of cartilage lesion gradually decreased in one of them.Follow-up study showed the outcomes had negative correlation with age and K-L grade (P ＜0.05),but no certain correlation with BMI in PRP group (P ＞ 0.05).Clinical effects in both groups were decreased over time.Conclusions Intraarticular injection of PRP benefits to pain relief,decreased inflammation and tissue repair,and has much better outcome in patients with younger age and lower K-L grade.However,BMI is not associated with the outcome.

Objective To study the effect of different dialysis modalities on pruritus in elderly maintenance hemodialysis patients.Methods Totally 51 patients were randomly divided into hemoperfusion combined with hemodialysis group (HD+ HP),hemodiafiltration group (HDF) and hemodialysis group (HD).Plasma β2-microglobulin(β2-MG) and intact parathyroid hormone (iPTH) were measured by means of radio immunoassay at pre and post dialysis,4 weeks and 8 weeks after dialysis,cutaneous pruritus was scored.The remission rate of itching was calculated at 8 weeks after dialysis.The parameters were compared among different groups.Results The level of plasma β2-MG was lower in HD+HP group after dialysis than pre dialysis [(13.48±3.05)mg/L vs.(16.27±4.73) mg/L,t＝2.044,P＜0.05],at 4 weeks and 8 weeks after dialysis,its levels were decreased to (10.97±3.25)mg/L(t＝3.808,P＝0.002)and (6.47±2.35)mg/L(t＝7.650,P＝0.000),respectively.The levels of iPTH were also found decrease from(887.5 ± 242.7)ng/L to (688.3 ±223.4)ng/L(t＝3.384,P＝0.004)at 4 weeks and (467.2±102.5) ng/L(t＝6.578,P＝0.000) at 8weeks after dialysis in HD+HP group (all P＜0.01).There were differences of the levels of plasma β2-MG and iPTH at 4 weeks and 8 weeks after dialysis in HDF group (all P＜ 0.05),but no differences of the levels of plasma β2-MG and iPTH during every period were found in HD group(all P＞0.05).The scores of cutaneous pruritus were decreased from (21.17± 5.01) scores to (13.37±2.85) scores(t＝ 5.580,P＝0.000)at 4 weeks and (8.52±4.38) scores(t＝7.838,P＝0.000)at 8 weeks after dialysis in HD+ HP group,and also the scores at 4 and 8 weeks after dialysis in HDF group (all P＜0.01),but there were no significant differences of the scores during every period in HD group (all P＞0.05).The remission rate of itching was better in HD+ HP group than in HDF group [88.24％ (15/17 cases) vs.58.82％ (10/17 cases),x2＝14.44,P＝0.000],better in HDF group than in HD group 23.53％ (4/17 cases) (x2 ＝4.37,P＝0.037).Conclusions HD+HP is superior to HDF in efficiently clear β2-MG and iPTH,and relief cutaneous pruritus,but HD can poorly clear β2-MG and iPTH or relief itching.

Objective To explore the association of sedentary behavior and time with risk of metabolic syndrome (MS).Methods A total of 10 149 subjects were recruited from local residents aged ≥40 years old in Jiashan County by cluster-random sampling method.The data including physical activity,job,sedentary time,and sleep,etc.were collected.Height and body weight,waist circumference,blood pressure,blood glucose,blood lipid,etc.were determined.Logistic regression was used for correlation analysis.Results The prevalence of MS was 28.64％ in local residents over 40 years old in Jiashan.The rate of sedentary behavior in all subjects was 67.57％,with 3 h sedentary time on average.Compared with non-MS group,the rates of sedentary behavior and sedentary time were significantly higher in MS group (P＜0.01).Multivariate logistic regression analysis showed that sedentary behavior was independently associated with an increased risk of MS after adjustment for age,sex,body mass index (BMI),smoking,drinking,and sedentary time(OR=1.16,95％ CI 1.03-1.30,P=0.017).Increased sedentary time was associated with higher risks of hypertension,type 2 diabetes mellitus,and dyslipidemia (P ＜ 0.05 or P ＜ 0.01).Sedentary time ≥ 5 h/d independently increased the risk of MS (OR =1.15,95％ CI 1.01-1.31,P =0.034).Conclusions There is a high prevalence of MS in adults over 40 years old living in the eastern coastal rural area.MS and its related diseases are closely associated with sedentary behavior and its duration.