Objective:To compare the proliferative and periodontal specific differentiation abilities of induced pluripotent stem cells (iPSCs) at different passages,and to investigate whether long term culturing would have a negative influence on their proliferation and specific differentiation capacity,thus providing a theoretical basis for further in-depth research on periodontal regeneration and the possible clinical applications of iPSCs.Methods:IPSCs derived from human gingival fibroblasts at passages 5,10,15 and 20 were recovered and cultured in vitro.Their morphology and proliferation rates were observed respectively.We further induced the iPSCs at different passages toward periodontal tissue under the treatment of growth/differentiation factor-5 (GDF-5) for 14 days through the EB routine,then compared the periodontal differentiation propensities between the different passages of iPSCs by detecting their calcified nodules formation by Alizarin red staining and assaying their relative periodontal tissue related marker expressions by qRT-PCR and immunofluorescence staining,including bone related markers:osteocalcin (OCN),bone sialoprotein (BSP);periodontal ligament related markers:periostin,vimentin;and cementum related markers:cementum attachment protein (CAP),cementum protein 1 (CEMP1).The untreated spontaneous differentiation groups were set as negative controls respectively.Results:iPSCs at different passages all showed a high proliferative capacity when cultured in vitro and turned into a spindlelike shape similar to fibroblasts upon periodontal specific differentiation.All iPSCs formed typical calcified nodules upon GDF-5 induction by Alizarin red staining in comparison to their untreated controls.The relative calcium deposition at all passages had been significantly upgraded under the treatment of GDF-5 (P5:t =2.125,P =0.003;P10:t =2.246,P =0.021;P15:t =3.754,P =0.004;P20:t =3.933,P =0.002),but no significant difference in their calcium deposition were detected within passages 5,10,15 and 20 (periodontal differentiation:F =2.365,P =0.109;spontaneously differentiation:F =2.901,P =0.067).Periodontal tissue related marker expressions of iPSCs at all passages had also been significandy upgraded under the treatment of GDF-5 (P ＜ 0.05),but still,no significant difference in their expression levels of periodontal tissue related proteins were detected within passages (BSP:F =0.926 7,P=0.450;vimentin:F=0.917 1,P=0.455;CEMP1:F=2.129,P=0.1367).Conclusion:Our results preliminarily confirmed that long term culturing won't influence the proliferation capacity and periodontal specific differentiation propensity of iPSCs,as they can still proliferate and differentiate toward periodontal cells with high efficiency upon growth factor induction after continuous passaging.Therefore,iPSCs could be recognized as a promising cell source for future possible application in periodontal tissue regeneration.

Objective To explore the impact of genetic background on pancreatic p-cell first-phase secretion function with L-arginine (L-ARG) stimulation test.Methods Plasma insulin level was detected in 201 cases before and after L-ARG stimulation test.Among them, 61 cases were newly diagnosed type 2 diabetic patients with family history of diabetes ( FH + DM ) , 55 newly diagnosed type 2 diabetic patients without family history of diabetes ( FH - DM) ,31 with normal glucose tolerance and family history of diabetes ( FH + ) 54 with normal glucose tolerance but without family history of diabetes ( FH - ).Homeostasis model assessment ( HOMA) was used to estimate insulin resistance (HOMA-IR).Results It was premised that gender, age and BMI were similar among the 4 groups.(1)TC,TG,fasting plasma glucose,2h plasma glucose,fasting insulin and HOMA-IR in the two groups of newly diagnosed type 2 diabetic patients with or without family history of diabetes were significantly higher than those in the two groups of normal glucose tolerance with or without family history of diabetes.The multiples of the peak level and the base level of insulin secretion in the groups of newly diagnosed diabetes were significantly lower than those in the groups of normal glucose tolerance with and without family history(P <0.05).(2) Insulin secretion reached a peak in 2 minutes and started to decline in 4 minutes in all the four groups.( 3 ) The multiples of the peak level and the base level of insulin secretion in normal glucose tolerance group with family history of diabetes were 20.8% lower than those in the group without family history, being 7.27 and 9.18 respectively ( P < 0.05 ).(4)Two-minute peak insulin secretion, HOMA-IR and age in the newly diagnosed type 2 diabetic group with family history of diabetes was significantly lower than these in the group without family history ( P < 0.05 ).The multiples of the peak level and the base level of insulin secretion in the newly diagnosed type 2 diabetic group with family history of diabetes and that group without family history were 5.18 and 5.31 respectively and there was no significant difference between the two groups( P >0.05).(5) When the normal glucose tolerance subjects with family history of diabetes progressed to suffer from diabetes, the multiples of the peak level and the base level of insulin secretion declined 43.6% (P < 0.05) more than those in the subjects still with normal glucose tolerance without family history.Conclusion In the early course of diabetes, insulin resistance dose not function significantly, but genetic background make the first-phase secretory function of the p-cell to decline gradually and type 2 diabetes occurs easily.In the absence of genetic background, insulin resistance makes first-phase the secretion of insulin to decline relatively slow.

0.05), respectively; the costs were 640.50yuan and 1 151.21yuan, respectively; the cost-effectiveness ratios were 7.01 and 12.28, respectively; the incremental cost-effectiveness ratio of Group B versus Group A was 220.1. CONCLUSION: The sequential therapy of levofloxacin is preferable in the treatment of lower respiratory infection.

ObjectivesTo report pathological and genetic features of 6 Chinese families with Xlinked Charcot-Marie-Tooth disease type 1 ( CMTX1 ).Methods The index cases from 6 families with CMTX1 are males with onset of disease between 11 and 24 years old.All of them had distal leg muscle weakness,accompanied with areflexia and sensory loss in the feet.Additionally,the index 1 presented with recurrent encephalopathy and the index case 5 with cerebellar ataxia.Peripheral neuropathy was found in 12 family members,while other 7 members showed talipescavus and hyporeflexia.Sural nerve biopsies were performed in all index cases.Connexin 32(Cx32) gene was analyzed in the index cases,8 affected and 10unaffected family members as well as 50 healthy women control subjects.ResultsMild to moderate loss of myelinated fiber with axonal degeneration and regeneration clusters were found in all index cases. Thin myelin fibers were found in 5,small onion bulbs in 3 and inflammatory infiltrates in 2.Five novel mutations (I20T,I127F,D178G,A197V,403_404insT) and one L10L synonymous mutation were detected in the 6index cases and their affected family members.The same mutations,in heterozygous state,were detected in 4 female family members without clinical symptoms,but not found in 6 male unaffected family members.The same mutations were not found in healthy control subjects.ConclusionsThe CMTX1 patients in our study present predominantly axonal lesions.Frequent novel Cx32 gene mutations indicated that private mutations may be common in Chinese CMTX1 patients.

ObjectiveTo investigate expression distribution of mutant connexin 32 (Cx32) protein in human endothelial cells in patients with X-linked Charcot-Marie-Tooth disease type 1 ( CMTX1 ) .MethodsNerve biopsies were performed in 3 patients with CMTX1 and in 3 non-CMTX1 controls. Cx32 mutations of c. 379A ＞ T( I127F), c. 533A ＞ G(D178G) and c. 590C ＞ T(A197V) were identified in these 3 patients respectively.Immunofluorescent (IMF) staining of nerve blood vessel was processed with antibodies against Cx32, Yon Willebrand factor and Cx40. The mutant Cx32 was constructed in pEGFP-N plasmid (pEGFP-N1-Cx32) and was transfected in HeLa cells. Cx32 and GRP78, a marker of endoplasmic reticulum ( ER), were stained by IMF in HeLa cells to investigate expression of mutant Cx32. ResultsIn 3 control cases, Cx32 was visualized by IMF staining as dots along gap junction of vascular endothelial cells,and it was coexisted with Cx40.However, immunoreactivity of Cx32 in 3 patients was predominantly decreased and was not located in endothelial gap junction. The transfection of 3 Cx32 mutants into HeLa cells demonstrated thepathogenic changes.The cells withthemutationc. 379A ＞T found Cx32 accumulations in the cytoplasm; the cells with mutation c. 533A ＞G showed few staining positive dots surrounding the nuclear and the cells with c. 590C ＞ T showed dot-like expression of Cx32 both in the cytoplasmicand cell membrane. The mutant Cx32 was not overlapped with expression of the marker of ER.ConclusionsMutant Cx32 might cause dysfunction of endothelial gap-junctions due to the abnormal expression of Cx32 in level and location in the vascular endothelial cells of CMTX1 patients.

BACKGROUND:To decrease operation amount of finite element analysis and increase its clinical practice,previous studies explored the material properties and 10 kinds of material attributes were assigned,which met the requirements of finite element analysis.Moreover,it can be used to calculate bone density.OBJECTIVE:To explore a method for measurement of bone density based on three-dimensional reconstruction and finite element analysis.METHODS:A total of 11 specimens of femoral superior segment were selected.The mass of control group was firstly measured.The experimental groups were treated with thin-slice high resolution CT scan and three-dimensional reconstruction in Mimics 10.0,volume meshing in Ansys,assigned with 10,100 and 400 kinds of material attributes Mimics,exported to Ansys to calculat the volumes of the block elements of every types of material attributes.The mass and the density of the specimens was harvested according to the empirical formula concerning the gray value and the bone density.All results were treated with one-way ANOVA.RESULTS AND CONCLUSION:One-way ANOVA showed that there were no significant differences between control group and experimental groups assigned with 10,100 and 400 kinds of material attributes (P＞0.28),and there were no significantly among the experimental groups (P＞0.8).Results show that the method was able to measure the mass and the density of bone quantitatively,as well as the proportion between compact bone and cancellous bone;to assign 10 kinds of material attributes to three-dimensional model of femur could match the needs for measurements.The results can be used as an initial preparation for the unification of bone density and finite element analysis for osteoporosis.

To compare the early-phase secretion between islet α cells and β cells in subjects with normal glucose tolerance and to explore the influencing genetic factors on the function of islet cells.40 subjects with normal glucose tolerance and family history of diabetes ( FH+ ) and 55 healthy volunteers without family diabetes history ( FH ) were collected.Fasting and L-arginine stimulating insulin,glucagon,and tasting free fatty acid,as well as other indicators were determined in all subjects.Insulin resistance was evaluated by homeostasis model assessment for insulin resistance.After adjnsting for sex,age,and body mass index,the insulin secretion peak of the two groups reached both at two min,and began to decline at four min,the peak multiple of FH+group was significantly less than that of FH-group (7.29 vs 8.88,P＜0.05) ; glucagon secretion peak of both groups reached at two min and began to decline by four min.Fasting glucagon and peak multiple were not significantly different (P＞0.05) ; The ratio of fasting insulin to fasting glucagon of the two groups was without significantly difference ( P＞0.05 ).Under diabetes genetic background,the function of β cells decreases even in subjects with normal glucose tolerance.

China ; Cities ; Health Knowledge, Attitudes, Practice ; Humans ; Schools ; Sodium Chloride, Dietary ; Students

OBJECTIVETo explore the quantitative relationship between the intensity of psychosocial stress and the degree of overall health damages.

METHODSA multi-group case-control study was designed and implemented. The cases included two groups of out-patients (177) and in-patients (214) in a hospital in Jianyang city, and controls (587) were from the follow-up cohort in the same city. Three groups were studied on the following contents: general demographic characteristics, psychosocial factors and the degree of health damages including mental, physical, and social status. Major statistical analyses were as follows: ranks test, ANOVA, cluster analysis, multinomial logistic regression and ordered-logit regression.

RESULTSOrdered-logit regression model showed that the odds ratio of negative life-events on degree of health damages was 1.335 (P < 0.01). This result showed that there was a positive dose-effect relationship between the negative life-events score and overall health damages. The utility of social support to overall health had protective effect (OR = 0.513).

CONCLUSIONNegative life-events were the major risk factors to overall health, and there was a dose-effect relationship between negative events and health damages. Function of social support played a protective factor for health.

Analysis of Variance ; Case-Control Studies ; Cohort Studies ; Educational Status ; Health Status ; Humans ; Marital Status ; Social Class ; Stress, Psychological ; psychology

Objective:To investigate the characteristics of 18F-DCFPyL PET/CT imaging in castration-resistant prostate cancer (CRPC) patients with different PSA levels. Methods:The imaging and clinical data of 50 patients with CRPC who underwent 18F-DCFPyL PET/CT examination in Chinese PLA General Hospital from January 2018 to December 2020 were analyzed retrospectively. The average age was 72 (54-95) years old. Serum total PSA was 92.28(0.36-2000.00) ng/ml. According to the total PSA level, the patients were divided into low PSA group(total PSA ≤ 1 ng/ml, n=9), medium PSA group (1 ng/ml10ng/ml, n=23). According to the standardized evaluation standard of molecular imaging, the suspicious tumor lesions on 18F-DCFPyL PET/CT imaging were scored by molecular imaging PSMA(miPSMA), and the miPSMA score ≥2 was defined as positive lesions. According to the number of lesions displayed by 18F-DCFPyL PET/CT, patients were divided into oligofocal group (the number of lesions ≤3) and multiple lesions group (the number of lesions >3). The imaging characteristics of patients in different groups were summarized. Results:The 18F-DCFPyL PET/CT imaging results of 50 cases in this study were all positive, including oligofocal group (n=27) and multiple lesions group (n=23). Of the 30 patients with unresected prostate, 18 had local recurrence of the prostate, while the other 12 patients with unresected prostate and 20 patients with resected prostate had no signs of local recurrence. The oligofocal group showed local recurrence, regional lymph node metastasis or bone metastasis. Patients with multiple lesions showed multiple lymph nodes and/or bone metastasis with or without local recurrence. There were 9, 18 and 4 patients with oligofoci in low, middle and high PSA groups, respectively.There were 27 patients in the oligonucleogenous group, and 21 of the 22 patients receiving local treatment were effective. All 3 patients treated with systemic treatments were effective. PSA progressed in 2 untreated patients. In the multi-foci group of 23 patients, 6 of 9 patients treated with abiraterone were effective. Two patients treated with enzalumide were ineffective. One of the 4 patients with chemotherapy was effective. One of the two patients treated with 177 Lu-PSMA nuclide was effective. One case did not respond to treatment with 89SrCl 2. Radiotherapy failed in 2 cases. PSA progressed in 3 untreated patients. Conclusions:18F-DCFPyLPET/CT imaging has a high detection rate of lesions in patients with CRPC and has potential guiding significance for follow-up treatment. The number of lesions in CRPC patients with different PSA levels was different, and the patients with low PSA levels were mainly oligofoci.