Calcineurin inhibitor screening should benefit from the strengths of each method. This paper reviews three calcineurin inhibitor screening methods:yeast positive screening model, yeast reporter gene high through- put screening model and calcineurin activity detection method. We have compared the range of application, advantages and disadvantages of each method, and combined effect of multiple methods. The yeast positive screening model can prevent the false positiveness caused by bacteriostat by using growth zooms instead of inhibition zooms. The yeast reporter gene high through-put screening model is suitable for pilot screening among a wide range of samples and is more efficient than the other methods. The calcineurin activity detection method can directly report the inhibitory ratio to a specific target. This review is expected provide a model-selection reference for the development of novel low-toxic calcineurin inhibitors.

Objective To investigate the age-associated changes of electrophysiological properties of atrial muscle and the relationship between these changes and atrial fibrillation (AF). Methods According to their ages forty Wistar rats were divided into 4 age groups: young group, adult group, middle-aged group and old group (n=10 each group ). Hearts were isolated and perfused by Langendorff. MAP (monophasic action potential) of atrial muscle was recorded. MAPD (duration of MAP) and ERP (effective refractory period) at 400 ms of the stimulation cycle length, and MAPD at other different stimulation cycle lengths were measured in each group. Results At the stimulation cycle length of 400 ms, MAPD_(90) of right atrial muscle prolonged gradually from young group to old group (P

Objective To explore clinical characteristics of patients with Klinefelter′s syndrome(KS; 47,XXY).Methods 227 male patients with 47,XXY treated by artificial insemination with donor(AID)were included.Age, education, height, weight, body mass index(BMI), testicular volume, FSH, LH, testosterone(T), prolactin(PRL), estradiol(E2)were analyzed retrospectively.Results Of these patients, their height were(176.4±5.5)cm, weight(74.5±12.7)kg, BMI(23.89±3.66)kg/m2[77 of overweight(33.92%)and 34 of obesity(14.98%)], FSH(38.35±14.33)IU/L, LH(19.40±9.00)IU/L, T(132.00±194.50)ng/dl, E2(23.90±15.00)ng/L, PRL(10.50±8.20)μg/L, E2/T 0.21±0.80.Testicular volume had the positive association with the level of T(r=0.197, P=0.003).BMI had the negative association with the serum concentration of T(r=-0.284, P=0.000), while positive association with the E2(r=0.174, P=0.009)and ratio of E2/T(r=0.323, P=0.000).Age had no association with T, E2, and E2/T(P>0.05), but had negative association with the serum concentration of LH(r=-0.154, P=0.02)nd FSH(r=-0.196,P=0.03).The higher education group were older(P<0.01), while the level of T were lower(P<0.01).Conclusion In patients with Klinefelter′s syndrome(KS; 47,XXY), level of T may associate with testicular volume.T, E2, and the ratio of E2/T seem to associate with their height, BMI, and education level.

Objective To investigate the clinical efficacy of Wumei Wan (Fructus Mume Pills) for diabetic gastroparesis with the syndrome of cold-heat mixture. Methods Sixty-eight diabetic gastroparesis patients with the syndrome of cold-heat mixture were evenly randomized into treatment group and control group. On the basis of treatment for controlling the blood glucose level, the treatment group was given oral use of Wumei Wan and the control group was given oral use of Cisapride. After treatment for one month, the therapeutic effect of both groups was evaluated, and the changes of gastrointestinal symptoms, gastric emptying, 2-hour postprandial glucose(2hPG), glycosylated hemoglobin(HbA1c), and fasting plasma glucose(FPG) in both groups were also observed. Results (1)The total effective rate of the treatment group was 88.24%, and that of the control group was 67.65%, the difference being significant(P<0.05). (2 ) After treatment , the scores of gastrointestinal symptoms of nausea and vomiting, gastric fullness, anorexia, belching and acid regurgitation, abdominal and gastric pain, diarrhea, and constipation in the treatment group were markedly improved(P<0.05 or P<0 . 01 compared with those before treatment ). In the control group, symptom relief was shown in nausea and vomiting, anorexia, and diarrhea(P<0.05). The treatment group had better effect on relieving nausea and vomiting, abdominal and gastric pain, diarrhea, and constipation than the control group(P<0.05 or P<0.01). (3)After treatment, 2hPG, FPG and HbA1c levels were obviously decreased in both groups(P<0.05 compared with those before treatment) , and the decrease in the treatment group was superior to that in the control group (P<0.05). (4)After treatment, half gastric emptying time, full gastric emptying time, and the contraction of gastric antrum were improved in both groups(P<0.05), and the improvement of gastric emptying in the treatment group was superior to that in the cont rol group(P<0.05). Conclusion Wumei Wan has better effect for the treatment of diabetic gastroparesis than western medicine Cisapride.

Objective To explore the pathogenesis of Hirayama disease from juvenile cervical curvature and growth rate.Methods Totally 60 patients diagnosed with Hirayama disease (HD) from 2009 to 2011 in our hospital were included in the present study.Patient's height and growth rate 1-2 years prior to the onset of disease were recalled by patients and family members.Lateral cervical X-ray was examined,and cervical curvature was measured by Borden's method.Results All the patients were adolescents with onset age at 12-25 (17.0 ± 2.4) years old and peak age of onset at 15-18 [45 cases (75.0％)].Fifty-seven cases were male and 3 cases were female.Cervical MRI examination of the 60 cases showed that the spinal cord atrophy involving C4-C8 vertebral level.The C line values for cervical curvature by Borden's method of the patients was 2.6 (1.2,4.2) mm.Among 60 patients,57 of them were with abnormal cervical curvature.The average height growth rate 1 year prior to the onset was (7.1 ± 1.8) cm.Conclusions The clinical manifestations that featured in overgrowth in the first two years and abnormal cervical vertebra curvature are possible related with pathogenesis of HD.HD is possibly a cervical spinal cord compression disease,which is associated with cervical spinal dysplasia during juvenile growth.

Objective To evaluate the effect of combination therapy of tolterodine and tamsulosin in improving symptoms in patients with indwelling double-J ureteral stents.Methods A total of 96 patients underwent placement of a double-J ureteral stent after retrograde ureteroscopy for urinary stone disease,which were prospectively randomized ( random numbers table) into two groups.The tamsulosin group ( n =48) was administered tamsulosin 0.2 mg once daily,the combination group (n =48) was administcred tamsulosin 0.2 mg once daily and tolterodine 2 mg twice daily.All the patients completed a validated Ureteral Stent Symptom Questionnaire (USSQ),the Overactive Bladder Symptom Score (OABSS),the International Prostate Symptom Score (IPSS) one day before he placement and 4 weeks after stent placement of stent.Results The mean urinary symptom index score (25.4 ± 4.0 vs 15.3 ± 2.9,P ＜ 0.0001 ),the mean pain index score (15.0 ±2.6 vs10.3 ±2.1,P ＜0.0001),the mean general health index score (15.4 ±2.2 vs 11.0±1.5,P＜0.0001),the work performance index score (13.0±2.2vs9.7±1.9,P＜0.0001),the sex index score (3.6 ± 1.1 vs 2.2 ± 0.9,P ＜0.0001 ) improved significantly in combination group than those in tamsulosin group.There was a statistically significant difference between tamsulosin group and combination group in the IPSS score ( 12.8 ± 2.0 vs 9.2 ± 1.7,P ＜ 0.0001 ) and OABSS score ( 6.7 ± 1.4 vs 4.2 ± 1.4,P ＜ 0.0001 ) at the 4 weekfollow-up.Conclusions Indwelling ureteral stents have a significant impact on health related quality of life.It is effective for improving symptoms in patients with indwelling double-J ureteral stents by combination therapy with tolterodine and tamsulosin.

Objective To evaluate the effectiveness and security of Botulinum toxin type A (BTX-A) in treating interstitial cystitis /bladder pain syndrome,and also the effect of different site injection.Methods Sixty-nine consecutive BPS/IC patients from October 2011 to February 2016 were divided into three groups randomly,including control group (group A) and treatment group (group B and group C).There were 5 males and 64 females,with age from 23 to 66 years old (average 44.5 years old).Twentythree patients (1 male and 22 females,aged from 23 to 69 years old,with mean age of 44.6 years old) in control group (group A) underwent hydrodistention.Twenty-four patients (2 males and 22 females,aged from 27 to 65 years old,with mean age of 42.8 years old) in group B underwent BTX-A detrusor combined triangle injection plus hydrodistention.Twenty-two patients (2 males and 20 females,aged from 30 to 68 years old,with mean age of 44.3 years old) in group C underwent BTX-A triangle injection alone plus hydrodistention.The parameters such as daytime frequency,nocturia,visual analogue scale/score (VAS),functional cystemetric volume (FCV),post-void residual volume (PVR),QOL score,maximal bladder capacity (MBC),interstitial cystitis symtom index (ICSI),interstitial cystitisproblem index (ICPI),Qmax,and Hamilton anxiey scale (HAMA) score between pre-treatment and 1,3,6 and 9 months after therapy in the three groups were compared.Results There were no serious complications observed in the three groups.All parameters 1 month after therapy were obviously superior to that of pretreatment.The efficacy in control group decreased significantly 3 months after treatment except nocturia (3.0 vs.5.0),daytime frequency(7.0 vs.14.0)and Q (14.0 ml/s vs.13.0 ml/s).However,all parameters in research groups except PVR were still obviously superior to pretherapy.The parameters except Q and nocturia in group B had statistically significant difference from contemporaneous group A (all P ＜ 0.05).However,the parameters except Q nocturia and somatic anxiety score in group C had statistically significant difference from contemporaneous group A (all P ＜ 0.05).VAS (2.0 vs.3.0) and somatic anxiety score (6.0 vs.10.0) in group B were superior to group C (all P ＜ 0.05).When it came to 6 months after therapy,urinary frequence and urgency and pain symptoms were the same to pretherapy and all parameters had no statistically significant difference between pretherapy and after therapy in group A(all P ＞ 0.05).But all parameters except PVR in research group was still superior to pretherapy.the parameters except PVR,Q and QOL in group B had statistically significant difference from contemporaneous group A (all P ＜ 0.05).However,The parameters except PVR,Q MBC,ICSI and QOL in group C had statistically significant difference from contemporaneous group A (all P ＜ 0.05).Meanwhile,efficacy in group B was better than group C in term of ICPI (6.0 vs.8.0) and somatic anxiety score (7.0 vs.10.0) (all P ＜ 0.05).The efficacy decreased significantly 9 months after treatment in both group B and C,with no statistically significant difference compared with that of pretreatment.ICSI(10.0 vs.13.0),ICPI(9.0 vs.13.0),QOL(5.0 vs.6.0)in group B,and QOL(5.0 vs.6.0)in group C had statistically significant difference compared with the contemporary parameters in group A.ICSI(10.0 vs.12.0),MBC(285.0 ml vs.237.5 ml) in group B was better than that in group C (P ＜ 0.05).Conclusions Symptoms in IC/BPS patients can be alleviated significantly by detrusor BTX-A injection plus hydrodistention.Quality of life can be improved remarkably and HAMA scores can be reduced significantly after treatment.Thus,it's an effective therapeutic mnethod for IC/BPS,and detrusor combined triangle injection can provide a better effect than single triangle injection.

Objective To investigate the frequency of JAK2 V617F mutation and JAK2 V617F mutation allele burden in patients with essential thrombocythemia (ET), and explore the relationship between mutation and hematological parameters and coagulation function. Methods The clinical and laboratory parameters of 90 ET patients were analyzed. JAK2 V617F mutation was detected by AS-PCR and the mutation allele burden of JAK2 V617F was detected by qPCR. The correlation between mutation frequency and mutation burden of JAK2 V617F and blood laboratory parameters were investigated in ET. Results JAK2 V617F mutation was found in 50 patients (55.6 %). RBC [(4.67±0.89)×109/L vs (4.04±0.99)×109/L, P =0.003], WBC (11.64±5.20)×109/L vs (9.11±4.11)×109/L, P = 0.014], HCT (0.41±0.07) vs (0.36±0.07), P =0.005) in the JAK2 V617F mutated group were higher than those in the wild-type group. PT in mutated patients was longer than that in wild-type group [(13.18±1.63) s vs (12.02±1.24) s, P = 0.000]. The JAK2 V617F mutation allele burden was (29.91 ±18.63) %. No significant correlation was found between JAK2 V617F mutation allele burden and hematological parameters such as WBC, RBC and Plt (all P>0.05), but the JAK2 V617F mutation allele burden had a significant correlation with FDP (r = 0.456, P = 0.001). Conclusions JAK2 V617F mutation occurs in significant percentage patients with ET. Detection of JAK2 V617F mutation allele burden at diagnosis may play an important role in the early prevention of vascular events.

Objective To observe the clinical curative effect of cinacalcet combined with activated vitamin D for treatment of patients with secondary hyperparathyroidism (SHPT) undergoing maintenance hemodialysis (MHD).Methods Eighty-six patients with SHPT undergoing MHD admitted to the Blood Purification Center of Wuhan General Hospital of Chinese People's Liberation Army from April 2014 to April 2016 were enrolled, and they were divided into an observation group and a control group by random number table, 43 cases in each group. The patients in control group were given cinacalcet whose initial dose was 25 mg/d and maximum dose should not exceed 75 mg/d, and the calcium acetate orally; on the basis of control group, the patients in observation group were additionally given activated vitamin D therapy, and both groups were treated for consecutive 12 weeks. After treatment, the clinical therapeutic effect, serum calcium, serum phosphorus, calcium phosphorus product, intact parathyroid hormone (iPTH) levels and the incidence of adverse reactions were compared between the two groups.Results The total effective rate in observation group was higher than that of the control group [90.70% (39/43) vs. 74.42% (32/43),P < 0.05]. After treatment, the difference of the serum calcium, calciumphosphorus product were higher than those before treatment in both groups [serum calcium (mmol/L): the control group was 2.24±0.25 vs. 1.99±0.26, observation group was 2.60±0.21 vs. 2.03±0.24; calcium phosphorus product (mmol2/L2): the control group was 4.05±0.34 vs. 3.79±0.35, observation group was 4.25±0.37 vs. 3.86±0.36, allP < 0.05], serum phosphorus, iPTH were lower than those before treatment in both groups [phosphorus (mmol/L): the control group was 1.69±0.14 vs. 2.09±0.12, observation group was 1.15±0.18 vs. 2.03±0.16; iPTH (ng/L): the control group was 297.36±59.73 vs. 499.54±69.32, observation group was 198.53±57.32 vs. 492.92±67.54, allP < 0.05], the degrees of changes in observation group were more significant than those in control group [serum calcium (mmol/L): 2.60±0.21 vs. 2.24±0.25, serum phosphorus (mmol/L): 1.15±0.18 vs. 1.69±0.14, calcium phosphorus product (mmol2/L2): 4.25±0.37 vs. 4.05±0.34, iPTH (ng/L): 198.53±57.32 vs. 297.36±59.73, allP < 0.05]; and the incidence of adverse reactions was significantly lower in observation group than that of the control group [4.65% (2/43) vs. 20.93% (9/43),P < 0.05].Conclusion Cinacalcet combined with activated vitaminD for treatment of SHPT patients undergoing maintenance hemodialysis shows obvious curative effect, reduces the whole segment of iPTH, and simultaneously has less adverse reactions.

Objective:To observe the expressions of SENP1, SENP2and SENP6proteins in human malignant glioma tissue and cells, and to elucidate the their effects in the development of malignant glioma.Methods:The samples of normal human brain tissue and malignant glioma tissue were obtained and used as normal control group and malignant glioma group, respectively.The Cos7cells and the malignant glioma LN443and U343cells were cultured;the Cos7cells were used as normal cell control group, and the LN443and U343cells as malignant glioma cell group.Western blotting method was used to detect the expression levels of SENP1, SENP2and SENP6proteins in human malignant glioma tissue and cells.Results:In brain tissue, the expression levels of SENP1, SENP2and SENP6proteins in malignant glioma group were higher than those in normal control group (P＜0.05) .Compared with normal cell control group, the expression levels of SENP1, SENP2and SENP6proteins in the LN443and U343cells in malignant glioma cell group were significantly increased (P＜0.05) .Conclusion:SENP1, SENP2and SENP6proteins highly express in the malignant glioma tissue and cells, and they may play an important role in promoting the occurrence of malignant glioma.