OBJECTIVETo determine the effect of type I transmembrane protein (IRE1alpha) deletions on the cell cycle of human periodontal ligament fibroblasts (hPDLFs) cells.

METHODSBased on the IRE1alpha deletions, a full-length model was successfully constructed. Moreover, overlapping polymerase chain reaction mutagenesis facilitated the establishment of two deletion mutants of IREla (pD-Kinase, pD-Rnase). The full-length model and two mutant eukaryotic expression vectors were transfected into hPDLFs cells. Western blot analysis was performed to identify the expression in the cells. The changes in the cell cycle of hPDLFS cells were detected by flow cytometry (FCM).

RESULTSThe two deletion mutants of IRE1alpha with eukaryotic expression vectors were successfully constructed and correctly expressed in hPDLFs cells based on Western blot analysis. Under stress conditions, the FCM assay showed that cell percentage of S phases increased, whereas that of G1 phases decreased in the IRE1alpha group (P < 0.05) compared with the control group of tunicamycin (TM) treatment. Moreover, the cell percentage of the S phases decreased, whereas that of the G1 phases increased in the D-Rnase group (P < 0.05) compared with the control. The deletion mutant D-Kinase had no influence on hPDLFS cell proliferation and cycle (P>0.05).

CONCLUSIONUnder stress conditions, IRE1alpha can improve the cell cycle of hPDLFs cells from the G1 to the S phase. The deletion mutant D-Rnase cause hPDLFs cell growth arrest at the G1 phase, whereas deletion mutant D-Kinase has no significant effect.

Objective To explore characteristics of the elements of syndrome, the disease position and the relationship between chemical indicators and TCM syndromes of type 2 diabetic encephalopathy. Methods 2 501 cases of type 2 diabetes clinical data were collected from Guang'anmen Hospital of China Academy of Chinese Medical Sciences, Beijing University of Chinese Medicine, Dongzhimen Hospital, Dongfang Hospital, etc. in nearly 3 years, among which, 342 cases were type 2 diabetic encephalopathy. The original clinical data were double entried in epidata by two people, establishment forms in excel, factor analysis and Bayesian networks were used as data mining research methods. Results 20 elements which characteristic root more than 1 were derived by factor analysis, 68.4% were covered. Of all 20 elements, five factors belong to Yin, five factors belong to blood stasis; lassitude, shortness of breath, stool frequency were appeared when fasting glucose abnormalities; lassitude, hemiplegia were appeared when 2-hour postprandial blood glucose abnormalities;lassitude, feverish palms and soles, stool frequency, more nocturnal enuresis when glycated hemoglobin abnormalities by Bayesian networks. Conclusion The Elements of the syndrome of type 2 diabetes encephalopathy were deficiency of Yin and blood stasis; and the main positions for diabetic patients were liver, spleen and kidney. Patients with impaired fasting glucose were Qi deficiency; Patients with impaired 2-hour postprandial glucose were Qi deficiency or pathogenic wind attacking collaterals; Patients with abnormal hemoglobin were Qi deficiency and Yin deficiency.

Purpose To compare the difference of the genotype and allele of endothelial cell protein C receptor (EPCR)gene rs9574 C/G between Guangxi population and other ethnic groups.Methods The rs9574 C/G polymorphisms of EPCR in 130 cases of Guangxi population were detected by PCR and DNA sequencing.The distribution frequency of allele and genotype was compared with the other four ethnic groups (HapMap-CEU,HapMap-HCB,HapMap-JPT,HapMap-YRI),which was published by the human genome project.Results Three genotypes of CC,CT and TT were found in rs9574 C/G with the frequencies of 39.2％,46.2％,14.6％ respectively.the allele frequencies of C,T were 62.3％ and 37.7％.No significant difference was observed in the frequency of genotype and allele between male and female (P ＞ 0.05).There were significant differences in the genotype distribution among Guangxi population,HapMap-CEU and HapMap-YRI (P ＜ 0.05).Significant differences of allelic frequency were found among Guangxi population,HapMap-CEU,HapMap-JPT and HapMap-YRI.Condusion The polymorphisms of rs9574C/G in 3'-noncoding region of EPCR gene in Guangxi population were different in different regions and ethnic groups.

Objective To explore the relationship among the disease - syndrome - formula of type 2 diabetes.Methods Based on the clinical data of 2501 cases of type 2 diabetes, to study the relationship among the disease - syndrome - formula of type 2 diabetes by making use of association rules.Results Three formulas, Shengmai Yin (Pulse-activating Fluid), Liuwei Dihuang Decoction (Decoction of Six Herbs Including Rehmannia), and Sijunzi Decoction (Decoction of Four Noble Herbs), were selected from 122 ones to analyze the relationship between the formula and symptoms. The occurrence of fatigue with weakness, dry mouth with thirst, frequent urination with increased volume of urine, numbness of limbs, blurred vision, thirst with preference of drinking water, frequent nocturnal urination, and shortness of breath with inactive talking was in high frequency in those patients who were prescribed Shengmai Yin. The occurrence of fatigue with weakness, thirst with preference of drinking water, blurred vision, frequent nocturnal urination, dry mouth with thirst, numbness of limbs, shortness of breath with inactive talking, and soreness and weakness of lumbus and legs was in high frequency in those patients prescribed Liuwei Dihuang Decoction, and there existed a multi-association among these symptoms. The occurrence of fatigue with weakness, soreness and weakness of lumbus and legs, frequent nocturnal urination, shortness of breath with inactive talking, numbness of limbs, insomnia, and blurred vision was in high frequency in those patients who were prescribed Sijunzi Decoction, and there existed a multi-association among these symptoms.Conclusion The association rule can be adopted to analyze the relationship between formula and syndrome, and to explore the multi-association relationship among the symptoms of type 2 diabetes.

Objective To evaluate the effect of preoperative anxiety on the dosage of opioid drugs in perioperative period.Methods Fourty female patients who aged 18-65 years,ASA Ⅰ or Ⅱ, undergoing laparoscopic hysterectomy were enrolled during November 2014 to August 2015.Demo-graphic characteristics of the patients were recorded.The patients were separated into two groups ac-cording to their state anxiety inventory (SAI).The patients with SAI>37 score were included in the high-anxious patients group (group H,n=22)while the other patients with SAI≤37 were enrolled in the low-anxious patients group (group L,n=18).All patients received the SAI test at the day be-fore surgery.The anesthesia time,intraoperative remifentanil consumption,duration of the first time giving analgesia after surgery,consumption of sufentanil at 1(T1 ),2(T2 ),4(T3 ),6(T4 ),24(T5 ) and 48 hours(T6 )after operation were recorded.Visual analogue scale (VAS)scores,the times of pressing PCIA and the side effects of opioid 48 h after the surgery were recorded.Results In group H, the time for first dose of opioid after surgery was significantly shorter than group L (P <0.05).The con-sumption of sufentanil in group H was significantly more than group L at T1-T6 (P <0.05).The times of pressing PCIA at the T5 time point after operation in group H was more than group L (P<0.05).No sta-tistically difference was found in the incidence of nausea or vomit between the two groups.There were no re-spiratory depression, pruritus or urinary retention in all patients.Conclusion Patients with high preoperative anxiety level perceive needed more opioids in postoperative pain control.

OBJECTIVE To observe the regulation effect of chidamide on energy metabolism in HCT-8 and HT-29 cells. METHODS HCT-8 and HT-29 cells were treated with chidamide 5,10 and 20 μmol · L-1. Morphological changes of these cells were observed under an ordinary optical microscope. Cell proliferation was detected by MTT. ATP production was determined by CellTiter-Glo? assay kit. Metabolic changes were tested by glycolytic stress kit. The mRNA level of lactate dehydrogenase A (LDH-A)was analyzed by real-time quantitative PCR,whereas the protein level of LDH-A was analyzed by Western blotting. RESULTS Compared with control group,cell morphology of HCT-8 and HT-29 cells in chidamide treated group was irregular,accompanied by deformation,shrinkage and cell debris, and the inhibitory rate of proliferation increased(P<0.05). There was no significant difference in ATP total content between chidamide 5 and 10 μmol · L-1 16 h treatment groups,but in chidamide 20 μmol · L-1 treatment group it was decreased(P<0.05). Chidamide 20μmol · L-1 had no effect on oxygen consumption rate, but glycolysis ATP generation rate was reduced by 30.7% and 37.9%(P<0.05),respectively. Chidamide 20μmol · L-1 had no effect on LDH-A mRNA level,but it decreased the protein level of LDH-A(P<0.01). CONCLUSION Chidamide can abate the respiratory metabolic ability of HCT-8 and HT-29 cells. The mechanism may be related to the down-regulation of LDH-A.

Objective To investigate the effects of early intensive therapy on P cell function and long-term glycemic control in newly diagnosed type 2 diabetic patients with different recruiting fasting plasma glucose (FPG) levels.Methods A total of 382 newly diagnosed type 2 diabetic patients with FPG 7.0-16.7 mmol/L were randomly assigned to therapy with insulin in the form of continuous subcutaneous insulin infusion (CSII) or multiple daily injection (MDI) or oral hypoglycemic agents (OHA, by using gliclazide and/or metformin) for initial rapid correction of hyperglycemia.The treatments were stopped after euglycemia had been maintained for 2 weeks.The patients were followed longitudinally on diet alone for 1 year.Intravenous glucose tolerances tests (IVCTTs) were performed and blood glucose, insulin and proinsulin were measured before and after therapy as well as at 1-year follow-up.Homeostasis model assessment ( HOMA) of β cell function and insulin resistance index ( HOMA-β and HOMA-IR ) were calculated.All the patients were stratified on the recruiting FPG: stratum A (7.0 mmol/L≤ FPG < 11.1 mmol/L) , stratum B (11.1 mmol/L≤ FPG ≤ 16.7 mmol/L).Results More patients in stratum A achieved target glycemic control (94.4% vs 89.8% ) and in shorter time [(5.9 ±3.8)d vs(6.9 ±3.6)d, P <0.05] as compared with those in stratum B.B cell function represented by HOMA-β and acute insulin response ( AIR) improved significantly after intensive interventions in both stratum A and B patients.However, the remission rate at 1 year was significantly higher in stratum A patients (47.8% ) than those in stratum B (35.7%, P < 0.05).The patients treated with insulin (especially with CSII) had higher remission rates and better improvement of AIR at 1 year follow-up irrespective of the recruiting FPG (CSII or MDI vs OHA: 57.1% , 51.8% vs 32.8% in stratum A, P <0.05; 44.4% , 38.7% vs 18.6% in stratum B, P <0.05).Conclusions Compared with OHA, early short time intensive insulin treatment had more favorable outcomes on maintaining AIR and prolonged glycemic remission in newly diagnosed type 2 diabetic patients irrespective of the recruiting FPG levels.

Objective To evaluate the cost?benefit and cost?effectiveness of current strategy for preventing mother?to?child transmission (PMTCT) of hepatitis B virus. Methods A decision tree model with the Markov process was developed and simulated over the lifetime of a birth cohort in Zhejiang Province in 2016. The current PMTCT strategy was compared with universal vaccination and non?vaccination. Costs were assessed from social perspective. Benefits were the savings from reduced costs associated with disease and effectiveness were measured by quality?adjusted of life?years (QALY) gained. The net present value (NPV), cost?benefit ratio (BCR) and incremental cost?effectiveness ratio (ICER) were calculated. Univariate and Probabilistic Sensitivity Analyses (PSA) were performed to assess parameter uncertainties. The parameters of costs and utilities value of hepatitis B?related disease came from the results of the field survey, which were obtained by face?to?face questionnaire survey combined with inpatient medical records, including eight county and municipal hospitals in Jinhua, Jiaxing and Taizhou. A total of 626 outpatients and 523 inpatient patients were investigated. The annual total costs of infection was calculated by combining the costs of outpatient and inpatient. Results The PMTCT strategy showed a net?gain as 38 323.78 CNY per person, with BCR as 21.10, which was higher than 36 357.80 CNY per person and 13.58 respectively of universal vaccination. Compared with universal vaccination, the PMTCT strategy would save 2 787.07 CNY per additional QALY gained for every person, indicating that PMTCT would be cost?saving. The most important parameters that could affect BCR and ICER were the vaccine coverage rate and costs of hepatitis B related diseases respectively. The PSA showed the PMTCT strategy was preferable as it would gain more QALY and save costs. Conclusions The PMTCT strategy appeared as highly cost?beneficial and highly cost?effective. High vaccination rate was a key factor of high economic value.

Objective:To explore the performance of agitated saline contrast echocardiography(ASCE)in patients with patent foramen ovale(PFO)and the relationship of that with the prognosis of treatment of interventional closure.Methods:A total of 70 patients with PFO who were hospitalized in Haikou People's Hospital from October 2020 to August 2022 were selected as research objects.All patients were treated with interventional closure,and the preoperative and postoperative ASCE performances,as well as the ASCE grading were recorded.All patients were followed up for 12 months after operation,and then they were assessed to confirm whether there was any residual diversion.After that,the changes of Headache Impact Test-6 Scale(HIT-6)score between before and after operation were compared.Results:In the ASCE image characteristics and the morphology of the foramen ovale of 70 PFO patients,22 cases(31.42％)were type Ⅰ patients,whose diameter of the foramen ovale was less than 1 mm,and the patent foramen ovale without closure appeared fine needle-like,and 28 cases(40.00％)were type Ⅱ patients,whose diameter of the foramen ovale was between 2 and 3mm,and the patent foramen ovale without closure appeared tunnel-like,and 20 cases(28.57％)were type Ⅲ patients,whose diameter of the foramen ovale was≥3mm,and the patent foramen ovale without closure appeared pouch-like.The results of re-examined ASCE of 70 patients indicated that the positions of the occluder were favorable in type Ⅰ,Ⅱ and Ⅲ patients,which closely attached to the atrial septum.There was not residual blood flow to pass through the atrial septum in all patients.Preoperative ASCE RLS grading showed that 20 cases(28.57％)were rest RLS grade 0,and 25 cases(35.71％)were grade I,and 14 cases(20.00％)were grade Ⅱ,and 11 cases(15.71％)were grade Ⅲ.The post classification of Waals maneuver indicated 30 cases(42.85％)were RLS grade I,and 25 cases(35.71％)were RLS grade Ⅱ,and 15 cases(21.4％)were RLS grade Ⅲ.Postoperative ASCE RLS grading showed that there was RLS in all patients,and the difference in RLS grading between before and after surgery was statistically significant(x2=85.783,P<0.05).In the 70 patients,ASCE results showed that the front-back diameter of left atrium of patients at 0 grade RLS was significantly less than that of patients at≥1 grade RLS,and the difference was statistically significant(t=8.783,P<0.05).There were no adverse events such as bleeding,infection,thromboembolism,stroke and occluder detachment occurred in all patients after surgery.The preoperative HIT-6 score of 70 patients was(73.85±5.79)points,and the scores decreased respectively to(50.82±6.57)and(39.06±4.69)points at the 1st and 6th months after surgery,and the difference of HIT-6 scores between them was significant(t=3.783,P<0.05).The results of re-examined ASCE at the 1st and 6th months after surgery showed that there was no RLS diversion phenomenon in 70 patients,and the results of trans thoracic echocardiography(TTE)indicated that RLS diversion existed in two patients.Conclusion:The diversion grade can be confirmed after PFO patients undergo the combined examination of ASCE,rest status examination and Waals maneuver,which has a certain of reference value in preoperative assessment for disease condition and in postoperative judgement for curative effect.

Objective To evaluate the cost?benefit and cost?effectiveness of current strategy for preventing mother?to?child transmission (PMTCT) of hepatitis B virus. Methods A decision tree model with the Markov process was developed and simulated over the lifetime of a birth cohort in Zhejiang Province in 2016. The current PMTCT strategy was compared with universal vaccination and non?vaccination. Costs were assessed from social perspective. Benefits were the savings from reduced costs associated with disease and effectiveness were measured by quality?adjusted of life?years (QALY) gained. The net present value (NPV), cost?benefit ratio (BCR) and incremental cost?effectiveness ratio (ICER) were calculated. Univariate and Probabilistic Sensitivity Analyses (PSA) were performed to assess parameter uncertainties. The parameters of costs and utilities value of hepatitis B?related disease came from the results of the field survey, which were obtained by face?to?face questionnaire survey combined with inpatient medical records, including eight county and municipal hospitals in Jinhua, Jiaxing and Taizhou. A total of 626 outpatients and 523 inpatient patients were investigated. The annual total costs of infection was calculated by combining the costs of outpatient and inpatient. Results The PMTCT strategy showed a net?gain as 38 323.78 CNY per person, with BCR as 21.10, which was higher than 36 357.80 CNY per person and 13.58 respectively of universal vaccination. Compared with universal vaccination, the PMTCT strategy would save 2 787.07 CNY per additional QALY gained for every person, indicating that PMTCT would be cost?saving. The most important parameters that could affect BCR and ICER were the vaccine coverage rate and costs of hepatitis B related diseases respectively. The PSA showed the PMTCT strategy was preferable as it would gain more QALY and save costs. Conclusions The PMTCT strategy appeared as highly cost?beneficial and highly cost?effective. High vaccination rate was a key factor of high economic value.