OBJECTIVE: To prepare simvastatin tablets and optimize its formulation by 3D printing technology. METHODS: A cylindrical tablet with a radius of 4 mm and a thickness of 4.8 mm were prepared by a 3D printer. The dosage of the adjuvant povidone 30(PVP K30), lactose/microcrystalline cellulose (MCC) and absolute ethanol were taken as an investigation factor, the cumulative dissolution rate of 5, 15 and 30 min were used as an evaluation index,and the central composite design-response surface method was used to optimize the formulation and verify, the effect of dissolution were researched by compared with the traditional preparation. RESULTS: The optimal formulation value: the ratio of PVP K30 was 30%, the mass ratio of lactose/MCC was 2.5, the ratio of absolute ethanol was 74%, and the cumulative dissolution in vitro of the printed tablets at 5, 15, 30 min was (45.7±0.5)%, (70.9±0.7)%, (89.7±2.3)% respectively, the deviation between the verification result and the predicted value were less than 5%. The preparation of dissolutioncurve is similar with commercially available formulations. CONCLUSION: The successful preparation of 3D printed simvastatin tablets is simple and the mathematical model is predictive.

OBJECTIVETo explore the impacts on the cognitive level of the children with mental retardation (MR) treated with JIN's three scalp needling therapy and the training for cognitive and perceptual disturbance so as to seek the more effective therapeutic method for the treatment of MR in children. Methods Sixty-six MR children were randomized into a head-point needle-retaining therapy + training group (group A) and a training after acupuncture group (group B). Seven cases and 12 cases were dropped in the two groups respectively. Twenty-six cases accomplished the treatment in the group A and 21 cases in the group B. In the group A, the points on the head were stimulated and the needles were retained (30 min after the general acupuncture, the needles on the body points were removed; and those on the head points were retained for 1 h, including Sishenzhen, Naosanzhen, Zhisanzhen and Niesanzhen). Simultaneously, the training for the cognitive perceptual disturbance was combined. In the group B, 1 h after needle retaining at the body points and head points, all the needles were removed; the training for the cognitive perceptual disturbance was followed. The treatment was given once a day, and the treatment for 3 months was required. Before and after treatment, the Wechsler intelligence scale for children (WISC) was used for evaluation and observation of verbal intelligence quotient (VIQ) , performance intelligence quotient (PIQ) and full-scale intelligence quotient (TFIQ) and score changes of 11 items such as information, picture vocabu; lary, arithmetic, picture generalization, comprehension, etc.

RESULTSAfter treatment, FIQ, VIQ and PIQ scores were different significantly as compared with those before treatment (all P<0. 01). In the group A, the results of picture vocabulary, animal egg laying, maze, block design and geometric figure were all improved significantly (all P<0. 05). In the group B, the results of information, comprehension, block design and geometric figure were all improved significantly (all P<0. 05). After treatment, concerning the value difference in FIQ and PIQ of the two groups; the changes in the group A were more significant (both P<0. 01). After treatment, the results of picture vocabulary and maze were improved significantly in the group A as compared with the group B (both P<. 01). The IQ categories changed apparently after treatment in the two groups, toward the higher level in tendency generally, but without significant difference (both P>0. 05).

CONCLUSIONThe simultaneous treatment with head point retaining of JIN's three needling therapy and the training for cognitive and perceptual disturbance obviously improves children patients' verbal comprehension, expression ability, hand-eye coordination ability, attention, logical reasoning ability and visual perception. The efficacy is better than that in the treatment of the training after acupuncture.

Acupuncture Therapy ; Child ; Child, Preschool ; Cognition ; Female ; Humans ; Intellectual Disability ; psychology ; therapy ; Intelligence Tests ; Male ; Perception ; Scalp ; Treatment Outcome

Objective By using primary cultured mouse renal tubular epithelial cells (TECs) to develop an in vitro model of simulated ischemia-reperfusion (IR) injury.Methods The outer medulla of C57BL/6J mouse kidney was flushed and primary cultured after digestion in type Ⅰ collagenase,and then immunocytochemical staining was used to verify TECs.Primary cultured TECs were immersed in mineral oil to simulate the ischemic process,and 60 min later the whole culture medium was added to simulate reperfusion process.The cells were collected and RAN was extracted at indicated time points after medium replacement.The expression of TNF-α,IL-1β and IL-6 was detected by using real-time fluorescence quantitative RT-PCR. The culture supernatants were collected at 24 h after medium replacement for detection of the expression of cytokine protein by using ELISA.Results Primary cultured TECs were identified by cobblestone-shaped morphology and then verified by cytokeratin 18 (CK18) staining.In TECs of IR group after medium replacement the mRNA expression of TNF-α,IL-1β and IL-6 was higher than in control group.The expression of TNF-α after medium replacement was increased to a peak level at 0.5 h,about (24.45 ±6.51) times (P＜0.01 ) higher than the control group,and gradually declined thereafter.The mRNA expression of IL-1β after medium replacement kept an increasing tendency,about ( 15.27 ± 4.29) times (P＜0.05) higher than the control group at 6 h,and that of IL-6 after medium replacement was increased to a peak level at 3 h,about ( 11.19 ±4.55) times (P＜0.01) higher than the control group. In the IR group at 24 h after medium replacement,the protein expression of NF-α,IL-1β and IL-6 in the supernatants was significantly higher than in the control group.Conclusion High purity of primary cultured TECs was achieved from the outer medulla of mouse kidney by separation and digestion.The in vitro model of simulated IR in primary cultured mouse renal TECs was successfully created using paraffin oil.

ObjectiveTo investigate the effect and underling mechanism of water-soluble CO-releasing molecules (CORM-3)on the alleviation of allograft rejectionafter mouse kidney transplantation.Methods A mice kidney transplantation model was established using C.FVB-Tg (Itgax-DTR/GFP)57Lan/J or C57BL/6J (H-2Kb) mice as donors,and Balb/c (H-2Kd) mice as recipients.After donor nephrectomy,kidney was preserved in UW solution which contained CORM-3 or iCORM (inactive CO-releasing molecules) for 24 h in 4℃.Recipient survival after removal of both na? ve kidneys,serum creatinine as well as graft histology was observed.In the C.FVB-Tg(ItgaxDTR/GFP) 57Lan/J donors,rDCs were acquired in vitro and selected by magnetic cell sorting (MACS) after graft nephrectomy.The expression of activation markers,CD80 and CD86,on rDC was assessed by using flow cytometry.ResultsThe graft medium survival time was 40.5 days in the iCORM group and 70 days in the CORM-3 group respectively (P＜0.05).CORM-3 preserved the graft function as shown by significantly lower serum creatinine (P＜0.05; or P＜0.01) and alleviated graft pathology injury.Diffuse infiltration of mononuclear cells in the interstitial tissues,moderate tubulitis and partial glomerular sclerosis were found in the iCORM graft kidney,while the CORM-3 graft kidney displayed almost normal histology.Meanwhile,CORM-3 suppressed the expression of CD80 and CD86 in donor-derived rDC.ConclusionCORM-3 can alleviate allograft rejection,prolong the graft survival,and improve kidney function in mouse kidney transplantation,probably via inhibiting rDC activation.

ObjectiveTo investigate the role of aldehyde dehydrogenase 2 (ALDH2) in the protection against tubular epithelial cells (TEC) ischemia/reperfusion (IR)injury induced by pretreatment with ethanol.Methods Mouse primary cultured TECs were pretreated with 50 mM ethanol 3 h before simulation of in vitro IR.Lactate dehydrogenase (LDH) release was assessed to evaluate the protection of ethanol pretreatment on IR injury.Thereafter,TECs were transfected with a negative control siRNA (NC) or an ALDH2-siRNA. The ALDH2 protein levels and ALDH enzymatic activities were assessed 48 h after transfection.Ethanol pretreatment and in vitro IR were performed on those transfected TECs.LDH release was assessed to evaluate the role of ALDH2 in the ethanol pretreatment-induced protection against IR injury.ResultsEthanol pretreatment significantly reduced the LDH release in TECs upon IR insult.As compared with NC group and INTERFERin group,the ALDH2 protein levels were decreased by 82.1％,ALDH enzymatic activities were decreased hy 67.3％,and the protective effect induced by ethanol pretreatment was almost completely abrogated in ALDH2-siRNA group.ConclusionEthanol pretreatment protects TECs against IR injury through ALDH2 dependent pathways.

OBJECTIVETo explore the mechanism of miR-124 inhibiting the proliferative activity of prostate cancer PC3 cells.

METHODSLuciferase reporter gene assay was used to examine the specific binding ability of miR-124 to PKM2 mRNA 3'-UTR. After miR-124 was transfected mimic to PC3 cells, the expression levels of PKM2 mRNA and protein were detected by real-time fluorescence quantitative PCR (qRT-PCR) and Western blot, respectively. The effects of miR-124 mimic and PKM2 siRNA on the proliferative activity of the PC3 cells were determined by MTT assay.

RESULTSThe expressions of PKM2 mRNA and protein were upregulated (5.12 +/- 0.35) times and (4.05 +/- 0.20) times respectively in the PC3 cells as compared with those in the RWPE-1 cells (P < 0.05). Luciferase reporter gene assay demonstrated that miR-124 targeted PKM2 3'-UTR. At 24 hours after transfection with miR-124 mimic, the PKM2 protein expression in the PC3 cells was downregulated (0.16 +/- 0.04) times (P < 0.05), while the PKM2 mRNA level was not changed significantly (P > 0.05), as compared with the control group. MTT assay showed that both miRNA-124 mimic and PKM2 siRNA could inhibit the proliferation of the PC3 cells, but the former exhibited a greater inhibitory effect than the latter. After transfection with miR-124 mimic and PKM2 siRNA, the cell growth rates were (66.20 +/- 5.10)% vs (82.10 +/- 6.35)% at 24 hours (P < 0.05) and (49.34 +/- 2.37)% vs (70.10 +/- 5.80)% at 48 hours (P < 0.05).

CONCLUSIONmiR-124 can suppress the proliferation of PC3 cells by regulating the PKM2 gene.

Carrier Proteins ; genetics ; metabolism ; Cell Line, Tumor ; Cell Proliferation ; genetics ; Humans ; Male ; Membrane Proteins ; genetics ; metabolism ; MicroRNAs ; genetics ; Prostatic Neoplasms ; genetics ; metabolism ; pathology ; Thyroid Hormones ; genetics ; metabolism ; Transfection

OBJECTIVE:To provide references for rational use of human serum albumin.METHODS:The inpatients receiving human serum albumin in the hospital during Jan.-Dec.2014 were investigated retrospectively and analyzed in respects of general information of patients,serum albumin level before medication,drug dosage,drug cost and reasons.RESULTS:Eight thousand seven hundred and sixty patients from top 8 departments in the list of the number of patients of human serum albumin were included in the study,accounting for 64％ of total.There were 5 244 male (59.9％) and 3 516 female (40.1％);patient's age mainly ranged ＞ 30-60 year-old (52.6％).The number of patients of albumin was the largest in general surgery department (2 588 cases) and cardio vascular surgery department (2 348 cases).Referring to domestic package inserts,8 210 patients (about 93.7％) were in accor dance with indications,and hypoproteinemia was the most common medication indication (35.6 ％).The drug dosage per capita of ICU was the highest (152.5 g);total drug cost was the highest in general surgery department (13 334 396.4 yuan);treatment course per capita of infection disease department was the longest (10.7 d).CONCLUSION:There is irrational use of human serum albumin in our hospital and difference of drug use among departments.It is necessary to strengthen human serum albumin use monitoring and establish rational medication evaluation system so as to promote standard drug use and realize rational distribution of medical resources.

Objective To investigate the effect of acupuncture combined with antidepressant on behavioral changes and 5-hydroxytryptamine 1A and 2A receptor ( 5-HT1AR and 5-HT2AR) mRNA expression in nucleus raphes dorsalis (NRD) of sleep deprivation depression (SDD) rats. Methods Eight normal Sprague Dawley (SD) rats were caged together without any stimulus or treatment, serving as the normal group. Thirty-two SD rats were given solitary raise for 21 days, together with chronic unpredictable mild stress ( CUMS) and rapid eye movement ( REM) sleep deprivation to establish the model. Twenty-eight rats completed the modeling successfully, and were divided into model group, acupuncture group, medicine group, and combination group, 7 rats in each group. Rats in the model group were given modeling treatment but without any intervention. Rats in the acupuncture group were given acupuncture on acupoint Yintang, Shenting and bilateral Taichong, acupuncture stimulation lasting 15 s every 10 min and acupuncture retention for 20 min in each day. Rats in the medicine group were given gastric gavage of Zoloft solution (0.83 mg·kg-1·d-1) and Alprazolam solution (0.067 mg· kg-1·d-1) , and rats in the combination group were given both acupuncture and medicine intervention. The intervention for the three intervention groups lasted for 7 days. Results Compared with the normal group, body weight, Open-field scores and sucrose preference of the model rats were significantly decreased on modeling day 7, 14, 21 (P<0.05). On modeling day 28, body weight and Open-field scores of each intervention group were significantly increased compared with those of the model group (P<0.05), and Open-field scores of combination group differed from those of acupuncture group and medicine group ( P<0.05). Only sucrose preference of combination group was improved significantly compared with the model group (P<0.05). Compared with the model group, NRD 5-HT1AR mRNA expression level was increased in the combination group (P<0.05), and NRD 5-HT2AR mRNA expression level was down-regulated in all of the three intervention groups ( P<0.05). Conclusion Acupuncture and antidepressant treatment covering 7 days can evoke rapid effect on SDD rats, which can improve the slow effect of antidepressant.

Objective To investigate the clinical significance of serum cytokines concentrations and A- PACHE scores in evaluating the illness state for critical trauma patients.Methods A clinical prospective self-control trial was performed,in which 36 patients admitted to ICU by SIRS were enrolled.Objects were divided into mild and severe trauma group according to APACHE score.The TNF-?and IL-6 concentrations were determined on the 1st, 3rd and 5th day of admission,the APACHE score were assessed at the same time.Statistic analysis was performed according to this group.Results The TNF-?concentrations decreased continuously in the following days while IL-6 decreased from the 7th day in the mild trauma group.In the severe trauma group the TNF-?and IL-6,APACHE score concentrations kept increasing.There was a significant difference of TNF-?and IL-6 concentrations between severe trauma and mild trauma group.Conclusion Dynamic measurement of TNF-?and IL-6 concentrations with APACHE score provide great help to evaluate the illness state and predict the prognosis.

Emerging evidence has indicated that circular RNAs (circRNAs) play pivotal roles in the regulation of cellular processes and are found to be aberrantly expressed in a variety of tumors.However,the clinical role of circRNAs in bladder cancer (BC) and the molecular mechanisms have yet to be fully understood.In this study,the clinical specimens were obtained and the expression level of a circRNA BCRC4 was detected by real-time PCR in both BC tissues and cell line.The circular RNA over-expression plasmid was constructed and transfected into BC cells and related cell line.The cell cycles and apoptosis were observed using inverted microscope and flow cytometry.Western blotting was used to compare the relative protein expression of groups with different treatments.It was found that circRNA BCRC4 expression was lower in BC tissues than in adjacent normal tissues.Furthermore,consequences of fomed-expression of BCRC4 promoted apoptosis and inhibited viability of T24T and UMUC3 cells,and up-regulated BCRC4-inereased miR-101 level,which suppressed EZH2 expression in both RNA and protein levels.In addition,gambogic acid (GA) is a promising natural anticancer compound for BC therapy,and GA treatment increased the BCRC4 expression in T24T and UMUC3 cells in a dose-dependent manner.Altogether,our findings suggest that BCRC4 functions as a tumor suppressor in BC,and mediates anticancer function,at least in part,by up-regulating the expression of miR-101.Targeting this newly identified circRNA may help us develop a novel strategy for treating human BC.