Objective: To effectively reduce the concentration of poisons in cleanroom, protect the health of workers, realize the optimization and automatic control of the new return air device. And the influence of initial concentration, air volume, temperature and relative humidity of formaldehyde on the purification effect of the new return air device was explored. Methods: The purification effect of the new return air device installed with the activated carbon and the photocatalyst purification net or ordinary activated carbon purification network was tested in a 60 m³ simulated cleanroom. The concentration of formaldehyde was determined by solution absorption-phenol reagent spectrophotometry. Based on the single factor experiment to determine the combination of two purification nets. The effects of air volume, initial formaldehyde concentration, temperature and relative humidity on the purification effect of the new return air device were investigated by orthogonal test. Then, the performance parameters of the return air device to purify formaldehyde were determined. Results: The formaldehyde purification efficiency of the two types of purification nets in the new return air device was higher than that of the ordinary activated carbon purification network (P<0.05) . The combination of activated carbon and photocatalyst purification net has no effect on the formaldehyde purification efficiency of the return air device (P>0.05) . According to the direct analysis and variance analysis, air volume was the most sensitive factor (F value is 18.894, P<0.05) , followed by initial concentration (F value is 16.128, P<0.05) , while temperature and relative humidity have little effect (F value is 0.041 and 0.599, respectively, P>0.05) . LSD analysis showed that there was no significant difference in the purification efficiency of formaldehyde between 475 m³/h and 626 m³/h (P>0.05) . From the perspective of formaldehyde purification efficiency and energy saving, when the air volume is set to 475 m³/h, the new return air device has higher purification efficiency for high concentration of formaldehyde. Conclusion The new return air device consisting of activated carbon and photocatalyst purification net can play a good purification role in cleanroom with different temperatures and different humidity. Its formaldehyde purification efficiency is affected by air volume and initial concentration.

OBJECTIVE:To establish the method for the identification of impurities in succinylcholine chloride raw material. METHODS:Two-dimensional UPLC-QTOF-MS was adopted. One dimension chromatographic condition was Hypersil GOLD C18 column using buffer solution(22 mmol/L sodium pentanesulfonate+50 mmol/L sodium chloride+5 mmol/L sulphuric acid)as mobile phase A and acetonitrile as mobile phase B,volume ratio of mobile phase A to mobile phase B 95:5,column temperature of 40 ℃,flow rate of 1.0 mL/min,detection wavelength of 214 nm. Two-dimension chromatographic condition was ACQUITY UPLC BEH C18column using 0.1% ammonia as mobile phase A and acetonitrile as mobile phase B(gradient elution)with column temperature of 30 ℃ at flow rate of 0.4 mL/min. Ionization mode was ESI+ with capillary voltage of 2.5 kV,source temperature of 120 ℃,temperature of atomizing gas at 450 ℃,flow rate of atomizing gas at 900 L/h,acquisition mode of MSE. RESULTS:The succinic acid,succinyl chloride(intermediate),pyridine(reagent)and other impurities were detected in succinylcholine chloride by one dimensional method of HPLC. Other 4 obvious unknown impurities were named impurity 1,impurity 2,impurity 3 and impurity 4,among which the apparent content of impurity 2 was the highest. Two-dimensional method of HPLC-QTOF-MS deduced that the impurity 2 was dehydrogenate succinylcholine chloride and impurity 4 was succinylcholine chloride. The impurity 1 and impurity 3 were not detected in mass spectrum. CONCLUSIONS:Establish method for the identification of impurity in succinylcholine chloride raw material,and the results of research are used for the evaluation of the quality of the succinylcholine chloride raw material.

Objective To explore the clinical value of nodal metastatic characteristics in predicting the distant metastasis of papillary thyroid carcinoma (PTC).Methods A total of 1 408 PTC patients who met the inclusion criteria and received initial thyroidectomy at our department from January 2006 to December 2011 were enrolled in this study.Results After a median follow-up time of 7.8 years,distant metastases developed in 46 patients.Patients with lateral neck lymph node metastasis ≥7,individual size of lateral neck lymph node metastasis ≥ 1.15 cm and the total number of cervical lymph node metastasis ≥9 were prone to higher risk of distant metastasis;the high risk group had a lower 10-year distant metastasisfree survival (78.7％ vs.98％,x2 =122.941,P ＜0.01) and a shorter distant metastasis-free survival time (99.2 M vs.122.5 M,x2 =122.941,P ＜ 0.01).Conclusions Lateral lymph node metastasis is an independent risk factor for distant metastasis in PTC patients.

Mitochondrial diseases are maternally inherited heterogeneous disorders that are primarily caused by mitochondrial DNA (mtDNA) mutations. Depending on the ratio of mutant to wild-type mtDNA, known as heteroplasmy, mitochondrial defects can result in a wide spectrum of clinical manifestations. Mitochondria-targeted endonucleases provide an alternative avenue for treating mitochondrial disorders via targeted destruction of the mutant mtDNA and induction of heteroplasmic shifting. Here, we generated mitochondrial disease patient-specific induced pluripotent stem cells (MiPSCs) that harbored a high proportion of m.3243A>G mtDNA mutations and caused mitochondrial encephalomyopathy and stroke-like episodes (MELAS). We engineered mitochondrial-targeted transcription activator-like effector nucleases (mitoTALENs) and successfully eliminated the m.3243A>G mutation in MiPSCs. Off-target mutagenesis was not detected in the targeted MiPSC clones. Utilizing a dual fluorescence iPSC reporter cell line expressing a 3243G mutant mtDNA sequence in the nuclear genome, mitoTALENs displayed a significantly limited ability to target the nuclear genome compared with nuclear-localized TALENs. Moreover, genetically rescued MiPSCs displayed normal mitochondrial respiration and energy production. Moreover, neuronal progenitor cells differentiated from the rescued MiPSCs also demonstrated normal metabolic profiles. Furthermore, we successfully achieved reduction in the human m.3243A>G mtDNA mutation in porcine oocytes via injection of mitoTALEN mRNA. Our study shows the great potential for using mitoTALENs for specific targeting of mutant mtDNA both in iPSCs and mammalian oocytes, which not only provides a new avenue for studying mitochondrial biology and disease but also suggests a potential therapeutic approach for the treatment of mitochondrial disease, as well as the prevention of germline transmission of mutant mtDNA.
Animals ; DNA, Mitochondrial ; genetics ; Humans ; Induced Pluripotent Stem Cells ; cytology ; metabolism ; MELAS Syndrome ; genetics ; Male ; Mice ; Microsatellite Repeats ; genetics ; Mitochondria ; genetics ; metabolism ; Mutation ; genetics

T2DM+LIRA group and T2DM+LIRA+UC-MSCs group (P<0. 05). The ratio of insulin positive area in pancreas tissue was obviously rised, while the ratio of glucagon positive area in pancreas tissue was clearly descended in T2DM+LIRA+UC-MSCs group. And the same difference in valuating islet cells apoptosis by TUNEL could be observed ( P<0. 05). The expression of NF-κB and TLR4 protein in pancreas tissue of T2DM+LIRA+UC-MSCs group were the least amongthefourgroups[(0.75±0.10)vs(0.60±0.08),(0.47±0.08),(0.31±0.04),P<0.05]and[(1.24± 0. 12) vs (0. 93 ± 0. 10), (0. 95 ± 0. 09), (0. 74 ± 0. 07), P<0. 05 ]. Conclusion The combined treatment of liraglutide with umbilical cord mesenchymal stem cells is superior over a single treatment of liraglutide or umbilical cord mesenchymal stem cells in improving the islet function in type 2 diabetes mellitus models, which may be related to the down modulating the TLR4/NF-κB inflammatory signaling pathway.

Objective To evaluate the efficacy and safety of human glucagon-like peptide-1 analogue liraglutide in newly diagnosed type 2 diabetes mellitus (T2DM) with glycosylated hemoglobin A1c (HbA1c) ＞ 9％.Methods This was an open-labelled,randomized,parallel-group,treat-to-target trial.Newly diagnosed T2DM patients with HbA1c ＞ 9％ were enrolled.These patients were treated with metformin with repaglinide and randomized to receive once-daily liraglutide (LIRA,n =25) or the insulin glargine (IGla,n =24) at bedtime.Efficacy and safety were assessed and compared after 18-month treatment.Results (1) Compared with the baseline,patients with LIRA had significantly reduced mean body weight,BMI and waist circumference (P ＜ 0.01),whereas,the above indexes were increased (P ＜ 0.01) in patients treated with IGla.(2) After 18 months of treatment,fasting plasma glucose (FPG),2-hour plasma glucose after a 75g oral glucose load(2hPG) and HbA1c were significantly improved in all patients (P ＜ 0.01),with 2hPG,mean blood glucose (MBG),the largest amplitude of glycemic excursions (LAGE),mean amplitude of glycemic excursions (MAGE) were significantly lower in LIRA group than in IGla group (all P ＜ 0.05).(3) HOMA-IR decreased in both groups (P ＜ 0.05).However,△I30/△G30,AUCCP180 and Matsuda index were only significantly increased in patients treated with LIRA (respectively,4.88 ± 1.55 vs 7.60 ± 1.91,9.23 ± 2.66 vs 13.18 ± 2.72,39.28 ± 20.35 vs 54.64 ± 23.34,all P ＜ 0.01),while HOMA-IR reduced(4.41 ± 1.58 vs 3.52 ± 1.44,P ＜0.05).But in IGla group only HOMAIR was reduced (4.92 ± 1.84 vs 4.57 ± 1.80,P ＜0.05).The index of △I30/△G30,AUCCP180 and Matsuda index in LIRA group are higher than those of indexes in IGla group(respectively,7.60 ± 1.91 vs 4.18 ± 1.00,13.18 ± 2.72 vs 10.53 ± 2.68,54.64 ± 23.34 vs 41.65 ± 17.84,all P ＜ 0.05),while HOMA-IR is lower (3.52 ± 1.44 vs 4.57 ± 1.80,P ＜ 0.05).(4) The rate of HbA1 c ≤ 6.5 ％ and the dosages of oral anti-diabetic drugs in LIRA group were significantly better than that in IGla group.(5) No significant differences were observed in hypoglycemic episodes and adverse events between two groups.Conclusion It seems that liraglutide is superior to insulin glargine in newly diagnosed T2DM patients with HbA1c ＞ 9％ in improving beta-cell function,insulin sensitivity and glucose homeostasis.

Objective To investigate the effect of liraglutide combined with bone marrow mesenchymal stem cells (BM-MSCs) on glucose metabolism in experimental type 1 diabetic (T1DM) rats.Methods T1 DM rats were established by injecting 60 mg/kg streptozotocin.According to random number table,they were divided into T1DM group (n =10),BM-MSCs group (n =10),LIRA group (n =10),and LIRA+BM-MSCs group (n =10),and were treated for 8 weeks respectively.Random blood glucose,24 h urine volume and protein excretion were tested.Serum concentrations of insulin,C-peptide,glucagon,gastrin,cholecystokinin,and glucagon-like peptid 1 (GLP-1) were assayed by ELISA.Expressions of insulin and glucagon in pancreas were measured by immunohistochemistry.Results After 8 weeks,the levels of random blood glucose,HbA1C,24 h urine volume and 24 h urinary protein excretion in group 4 were significantly decreased compared to the other three groups (P＜0.05).Compared to T1DM group and BM-MSCs group,the levels of insulin,C-peptide,gastrin,cholecystokinin and GLP-1 in LIRA+BM-MSCs group were significantly increased,while glucagon was decreased (P＜0.05).Compared to group LIRA,the levels of insulin,C-peptide,gastrin,and cholecystokinin in LIRA + BM-MSCs group were increased (P ＜ 0.05),there was no significantly difference in glucagon or GLP-1 (P＞0.05).The analysis revealed that the level of insulin was positively correlated with gastrin (r =0.544,P＜0.01),cholecystokinin (r =0.710,P＜0.01) and GLP-1 (r =0.669,P＜ 0.01),but was negatively correlated with glucagon (r =-0.506,P＜0.01);the level of glucagon was negatively correlated with gastrin (r =-0.364,P＜0.05),cholecystokinin (r =-0.433,P＜0.01) and GLP-1 (r =-0.591,P＜ 0.01).Compared to the other three groups respectively,immunohistochemistry displayed that the positive area of insulin in pancreas was significantly increased in LIRA + BM-MSCs group,while that of glucagon was decreased (P＜ 0.05).Conclusions By means of regulating gastrointestinal hormones efficiently,combination of liraglutide withBM-MSCs may improve glucose metabolism more efficaciously than treatment with a single agent in T1DM rats.

Objective To observe the effect and safety of the human glucagon-like peptide-1 analogue,liraglutide,versus insulin glargine in patients with type 2 diabetes mellitus inadequately controlled with metformin alone.Method Ninty patients with type 2 diabetes mellitus(aged 18-79 years,HbA1C 7.5％-10.0％,body mass index＜40 kg/m2) who had inadequate glycaemic control on metformin were allocated for the research with an open,randomized,parallel controlled clinical research method.The patients kept the original dose of metformin unchanged and were randomly assigned to the liraglutide group or the insulin glargine group according to a proportion of 1 ∶ 1.Liraglutide group started with a dose of 0.6 mg subcutaneous injection qd,changed to 1.2 mg subcutaneous injection qd after one week and kept unchanged until the end of the research.Insulin glargine group started with a dose of 0.1-0.2 U/kg according to the fingertips peripheral blood glucose level before breakfast on the continuous 3 d before every follow-up.At the baseline,after 4 weeks,12 weeks,20 weeks,and 26 weeks of treatment,HbA1C,blood glucose,lipids weight,blood pressure were arranged to measured.86 patients finally completed the study.Results Mean HbA1C and the success rate of HbA1C ＜7％ were similar between liraglutide group and insulin glargine group [(7.06 ± 0.87) ％ vs (7.25 ± 1.20) ％,47.73 ％ vs 45.23 ％,P＞0.05],while the percentage of subjects reaching the composite endpoint of HbA1C＜7％ with no hypoglycemia and no weight gain was significantly higher in liraglutide group than insulin group(P＜0.05) ; Fasting plasma glucose decreased more markedly in insulin glargine group,2 h postprandial plasma glucose was decreased more markedly in liraglutide group(P＜0.05 or P＜0.01).Liraglutide significantly reduced mean body weight by (3.21 ± 1.18) kg,waist circumference by (3.82 ± 1.21) cm,and body mass index by (1.95 ± 0.61) kg/m2 (P＜0.01 or P＜0.05),while in the insulin glargine group there sere rise of respective figure of(2.86 ± 0.43) kg,(1.52 ± 0.56) cm,and (0.61 ± 0.25) kg/m2 (P＜0.05),systolic blood pressure and serum triglyceride declined.There was no serious adverse affect in both groups,the incidence of mild hypoglycemia was significantly less in liraglutide group and has a statistically significant difference (4.55％ vs 21.43％,P＜0.05).Conclusions Liraglutide showed a good effect on reducing weight,systolic blood pressure,blood lipid and in addition to blood glucose control which is comparable to insulin glargine.What is more,liraglutide had good safety and tolerability,which can be regarded as a good choice for patients with type 2 diabetes mellitus inadequately controlled with metformin alone.

ObjectiveTo investigate the effects of liraglutide on the differentiation of human bone marrow mesenchymal stem cells (hBM-MSCs) into insulin-producing cells (IPCs).MethodsIn vitro,hBM-MSCs were induced into IPCs by three-stage induction procedure containing high glucose,nicotinamide,and liraglutide.The morphological change of cells was observed by inverted microscope during induction and the induced cells were confirmed by dithizone(DTZ) staining.The protein expressions of pancreatic and duodenal homeobox-1 (PDX-1),glucose transporter 2 (GLUT2),glucokinase(GK),and insulin in each stage of the induced cells were detected by Western blot.Insulin secretion was measured by ELISA.ResultsThe induced effect was pronounced after adding 10 nmol/L liraglutide for 7 days.Cells began to aggregate and get round gradually during induction,and the morphology of most cells appeared as grape-like aggregation and clustered islet-like cells by the end of induction.The number of DTZ positive cells and the protein expressions of PDX-1,GLUT2,GK,and insulin were increased gradually( P＜0.05 ).The basal and glucose-stimulated insulin secretion from induced cells was also increased gradually(P＜0.05).Conclusion BM-MSCs could be induced into IPCs by high glucose,nicotinamide,and liraglutide in vitro.

Objective To investigate the protective effect of quercetin on diabetic nephropathy and to explore its possible mechanism.Methods Type 2 diabetes mellitus rat model was established by feeding high-carbonhydrate-fat diet and injecting with streptozotocin.At 72 hour after injection,blood samples were collected from the tail veins of all rats.Those rats with blood glucose level ≥ 16.7 mmol/L were considered as the diabetes model been successfully established.The model rats were randomly divided into type 2 diabetic group ( group DM,n =9 ) and quercetin group ( group QUE,n =9 ).Other rats were used as normal controls (group NC,n =8).All rats were performed by intragastric administration for 8 weeks.At the end of experiment,the rats were sacrificed and fasting plasma glucose( FPG),fasting insulin( Flns),serum creatinine (SCr),blood urea nitrogen(BUN),TG,TC,LDL-C,24 h urine protein (24 h UP),and kidney index ( KI ) were evaluated.Pathological changes of kidney were observed by periodic acid-silver metheramine( PASM ).The expressions of ubiquitin and NF-κB p65 on glomeruli w ere examined by immunohistochemical method,and its association with the incidence of proteinuria was analyzed.Results In groups DM and QUE,the level of FPG [ ( 25.45 ± 1.23 ) mmol/L and ( 19.99 ± 1.20 ) mmoL/L],FIns [ ( 25.67 ± 2.58 ) mU/L and ( 19.29 ± 1.80 ) mU/L ],SCr[ ( 44.00 ± 2.53 ) μmol/L and ( 34.43 ± 2.23 )μmol/L],BUN[ ( 11.60 ± 0.39 )mmol/L and (8.20 ± 0.37) mmoL/L],TG [ (3.32 ± 0.22 ) mmol/L and (2.43±0.25)mmol/L],TC[(2.95 ±0.21) mmol/L and (2.24 ±0.17)mmol/L],LDL-C[(2.03 ±0.22 ) mmol/L and ( 1.49 ± 0.13 ) mmol/L ],24 h UP [ ( 46.67 ± 2.50 ) mg/24 h and ( 25.57 ± 2.82 )mg/24 h]and KI[ (9.76 ±0.30) × 103 and (8.44 ±0.26) × 103 ] were significantly increased than the indexes of group NC [ (6.56 ± 0.41 ) mmol/L,( 12.63 ± 1.41 ) mU/L,( 22.88 ± 2.36 ) μmol/L,( 5.45 ±0.51 ) mmoL/L,( 1.64 ± 0.1 1 ) mmol/L,( 1.33 ± 0.17 ) mmol/L,(0.46 ± 0.05 ) mmol/L,( 12.38 ±1.19)/24 h and (6.78 ±0.12) × 103].Moreover,the above indexes in group QUE were obviously lower than group DM.There was evidence of pathological changes associated with diabetes,such as focal and segmental sclerosis and thickened basement and mesangial expansion.The expressions of ubiquitin and NF-κB p65 in renal tissues of group DM increased significantly ( P ＜ 0.01 ).The expression of ubiquitin and NF-κB p65 were positively related with the level of 24 h UP ( r =0.893,0.879,P ＜ 0.01 ).Compared with group DM,all above indexes in group QUE were markedly alleviated ( P ＜ 0.01 ).The expression of ubiquitin and NF-κB p65 was reduced but didn't reach level in group NC ( P ＜ 0.01 ).Conclusion The increased expression of NF-κB induced by ubiquitin-proteasome system may participate in the pathogenesis of proteinuria in diabetic nephropathy.Quercetin has renal protective effects partly through reducing NF-κB p65 expression.