The development of a screening model for type 2 diabetes mellitus in community population of Xinjiang Karamay / 中华内分泌代谢杂志

Objective:To develop a nomogram model for screening of type 2 diabetes mellitus in a community population.Methods:From October to December, 2020, 6 028 community residents who participated in the " national health physical examination" in Karamay community with complete physical examination data and met the inclusion and exclusion criteria were selected. The physical examination data included medical history, physical examination, laboratory, and ultrasound reports. Random segmentation sampling was used to divide the population into modeling and validation cohorts, and LASSO regression analysis was used to screen for independent factors associated with diabetes diagnosis. The independent influencing factors were furthor incorporated into the multi-factor logistic regression, and the RMS software package was used to construct the column chart. The area under receiver operating characteristic(ROC) curve was used to measure the differentiation of the model. The calibration curve can directly reflect the calibration degree of the model.Results:In the modeling group, multivariate logistic regression analysis showed that age, gender(female), history of hypertension, history of hyperlipidemia, HbA 1C, urinary microalbumin, homeostasis model assessment for insulin resistance, and triglycerides and glucose index were independently associated with diabetes. OR were 1.053(95% CI 1.038-1.069), 0.681(95% CI 0.512-0.906), 1.802(95% CI 1.227-2.626), 1.789(95% CI 1.303-2.448), 10.973(95% CI 8.318-14.745), 1.002(95% CI 1.001-1.004), 2.914(95% CI 2.248-3.799), and 2.673(95% CI 2.03-3.536), respectively. The areas under ROC curves of the training set and the validation set were 0.945 and 0.955, respectively. The optimal critical value in the ROC curve was 0.178(sensitivity 0.930, specificity 0.839) in the training set and 0.201(sensitivity 0.945, specificity 0.848) in the validation set. Conclusion:The screening model of type 2 diabetes developed in this study has good accuracy, which can be used as a screening tool for high-risk population of type 2 diabetes.

A multicenter retrospective study on the real-world outcomes of autologous vs. allogeneic hematopoietic stem cell transplantation for peripheral T-cell lymphoma in China / 中华医学杂志(英文版)

BACKGROUND@#There were few studies on real-world data about autologous hematopoietic stem cell transplantation (auto-HSCT) or allogeneic HSCT (allo-HSCT) in peripheral T-cell lymphoma (PTCL). This study aimed to investigate the clinical outcomes of patients who received auto-HSCT or allo-HSCT in China.@*METHODS@#From July 2007 to June 2017, a total of 128 patients who received auto-HSCT (n  = 72) or allo-HSCT (n  = 56) at eight medical centers across China were included in this study. We retrospectively collected their demographic and clinical data and compared the clinical outcomes between groups.@*RESULTS@#Patients receiving allo-HSCT were more likely to be diagnosed with stage III or IV disease (95% vs. 82%, P = 0.027), bone marrow involvement (42% vs. 15%, P = 0.001), chemotherapy-resistant disease (41% vs. 8%, P = 0.001), and progression disease (32% vs. 4%, P < 0.001) at transplantation than those receiving auto-HSCT. With a median follow-up of 30 (2-143) months, 3-year overall survival (OS) and progression-free survival (PFS) in the auto-HSCT group were 70%(48/63) and 59%(42/63), respectively. Three-year OS and PFS for allo-HSCT recipients were 46%(27/54) and 44%(29/54), respectively. There was no difference in relapse rate (34%[17/63] in auto-HSCT vs. 29%[15/54] in allo-HSCT, P = 0.840). Three-year non-relapse mortality rate in auto-HSCT recipients was 6%(4/63) compared with 27%(14/54) for allo-HSCT recipients (P = 0.004). Subanalyses showed that patients with lower prognostic index scores for PTCL (PIT) who received auto-HSCT in an upfront setting had a better outcome than patients with higher PIT scores (3-year OS: 85% vs. 40%, P = 0.003). Patients with complete remission (CR) undergoing auto-HSCT had better survival (3-year OS: 88% vs. 48% in allo-HSCT, P = 0.008). For patients beyond CR, the outcome of patients who received allo-HSCT was similar to that in the atuo-HSCT group (3-year OS: 51% vs. 46%, P = 0.300).@*CONCLUSIONS@#Our study provided real-world data about auto-HSCT and allo-HSCT in China. Auto-HSCT seemed to be associated with better survival for patients in good condition (lower PIT score and/or better disease control). For patients possessing unfavorable characteristics, the survival of patients receiving allo-HSCT group was similar to that in the auto-HSCT group.

Correlation between sleep duration and incident diabetes among residents of different ages in Xinjiang region / 中华内分泌代谢杂志

Objective:To explore the correlation between sleep duration and incident diabetes among residents of different ages in Xinjiang region.Methods:A total of 9 541 residents, aged 40 and over in Karamay, Xinjiang were identified by a cluster sampling method. Physical examinations and biochemical test were performed, and the data on sleep duration and lifestyle were obtained using standardized questionnaires. The population was divided into three categories according to sleep duration: insufficient sleep(<6 h), ideal sleep(6-8 h), and long sleep duration(>8 h). They were further divided into 2 subgroups based on age at survey. Those who were younger than 60 years old were defined as the middle-aged group, and the rest as the elderly. Multivariate logistic regression analysis was used to explore the correlation between sleep duration and the risk of diabetes in different age groups.Results:There existed an approximate U-shaped relationship between total sleep duration and fasting blood glucose as well as HbA 1C. Fasting blood glucose and HbA 1C were relatively lower among those with ideal sleep duration. After multivariable adjustment, residents with insufficient sleep revealed a 35% increased risk of diabetes( OR=1.35, 95% CI 1.06-1.71) compared with those with ideal sleep duration. However, the risk of diabetes was not significantly increased in those with long sleep duration( OR=1.04, 95% CI 0.94-1.14). Furthermore, the additive risk of insufficient sleep was only significant in the middle-aged group( OR=1.37, 95% CI 1.02-1.84). Conclusions:Among residents of different ages in Xinjiang region, insufficient sleep is associated with an increased risk of diabetes, which is only significant in the middle-aged group.

Effect of metformin on miRNA expression in type 2 diabetic patients and potential targets / 中华内分泌代谢杂志

Objective:To investigate the effect of metformin on the microRNA (miRNA) expression and screen potential target with network pharmacology analysis in patients with type 2 diabetes.Methods:Fifteen patients with new diagnosed type 2 diabetes admitted to our hospital were selected, who received metformin during hospitalization and after discharge. The expression of serum matrix metalloproteinase (MMP)-9, transforming growth factor (TGF)-β1, and myocardial fibrosis related miRNAs were compared before and 6 month after metformin treatment. In addition, gene ontology (GO) and KEGG pathway enrichment analysis were applied to analyze differential expression miRNAs showing statistical significance. Meanwhile, the network figure was established to reflect the target gene messenger RNA (mRNA) corresponding to differentially expressed miRNA.Results:Compared with pre-medication, the serum level of MMP-9 was significantly decreased after treatment ( P<0.05). Besides, the expression of homo sapiens microRNA (hsa-miR)29a-3p, hsa-miR133a-5p, hsa-miR21-5p, hsa-miR30c-5p, and hsa-miR1-3p in patients with type 2 diabetes were dramatically down-regulated by metformin ( P<0.05 or P<0.01). Results of GO analysis and KEGG pathway enrichment analysis showed that differentially expressed miRNAs were mainly concentrated in endoplasmic reticulum lumen, synapse, basement membrane and other cell components. The molecular functions such as Rho GTPase binding and participation in extracellular matrix structural constituent were exerted through biological processes such as collagen catabolic process, regulation of short-term neuronal synaptic plasticity, and axon extension, which were mainly enriched in advanced glycation end products-receptor for advanced glycation end products (AGE-RAGE) signaling pathway in diabetic complications, tumor necrosis factor (TNF) signaling pathway, and Wnt signaling pathway, etc. The outcome of miRNA-mRNA network analysis demonstrated that there were 230 target genes mRNAs corresponding to differentially expressed miRNA. Conclusion:Metformin could play its role in the treatment of type 2 diabetes by down regulating the expression of miRNA, participating in the transduction of related cellular signaling pathways, regulating chromatin, nucleic acid binding, and enzyme activities.

Outcomes of 33 patients with anaplastic large cell lymphoma treated after hematopoietic stem cell transplantation / 中华血液学杂志

Objective: To explore the efficacy and prognostic factors of hematopoietic stem cell transplantation (HSCT) for the treatment of patients with anaplastic large cell lymphoma (ALCL) . Methods: The clinical records of 33 ALCL patients after HSCT were collected and analyzed retrospectively to evaluate the rates of overall survival (OS) and recurrence after autologous (auto-HSCT) and allogeneic HSCT (allo-HSCT) and the factors influencing prognosis. Results: The median-age of this cohort of 33 ALCL cases at diagnosis was 31 (12-57) years old with a male/female ratio of 23/10, 24 cases (72.7%) were ALK(+) and 9 ones (27.3%) ALK(-). Of them, 25 patients (19 ALK(+) and 6 ALK(-)) underwent auto-HSCT and 8 cases (5 ALK(+) and 3ALK(-)) allo-HSCT with a median follow-up of 18.7 (4.0-150.0) months. Disease states before HSCT were as follows: only 6 patients achieved CR status and received auto-HSCT, 16 patients achieved PR (14 cases by auto-HSCT and 2 ones allo-HSCT) , the rest 11 cases were refractory/relapse (5 cases by auto-HSCT and 6 ones allo-HSCT) . There were 7 cases died of disease progression (5 after auto-HSCT and 2 allo-HSCT) and 5 cases treatment-related mortality (TRM) (2 after auto-HSCT and 3 allo-HSCT) , TRM of two groups were 8.0% and 37.5%, respectively. Both the median progression-free survival (PFS) and OS were 15 months after auto-HSCT, the median PFS and OS after allo-HSCT were 3.7 (1.0-90.0) and 4.6 (1.0-90.0) months, respectively. There was no statistically significant difference in terms of survival curves between the two groups (OS and PFS, P=0.247 and P=0.317) . The 2-year OS rates in auto-HSCT and allo-HSCT groups were 72% and 50%, respectively. The 5-year OS rates in auto-HSCT and allo-HSCT groups were 36% and 25%, respectively. Conclusion: ALCL treated by chemotherapy produced high rates of overall and complete responses. Chemotherapy followed by auto-HSCT remained to be good choice for patients with poor prognostic factors. High-risk patients should be considered more beneficial from allo-HSCT.

Clinical outcomes of hematopoietic stem cell transplantation for angioimmunoblastic T-cell lymphoma / 中华血液学杂志

Objective: To evaluate clinical outcomes of autologous (auto-HSCT) and allogeneic hematopoietic stem cell transplantation (allo-HSCT) for angioimmunoblastic T-cell lymphoma (AITL) . Methods: From June 2007 to June 2017, clinical data of AITL patients who underwent HSCT in eight hospitals were assessed retrospectively. Results: Of 19 patients, 13 male and 6 female with a median age of 50 (32-60) years old, 12 auto-HSCT and 7 allo-HSCT recipients were enrolled in this study, all donors were HLA-identical siblings. Two of allo-HSCT recipients were relapsed auto-HSCT ones. There were 5 patients (5/12) in complete response (CR) status and 7 (7/12) in partial remission (PR) status before transplantation in auto-HSCT group, and 2 (2/7) in PR status and 3 (3/7) in progression disease (PD) status before transplantation in allo-HSCT group. The median follow-up for the surviving patients was 46.5 months (range, 1-100 months) for the whole series, two patients lost in auto-HSCT group. Three patients developed acute graft-versus-host disease (aGVHD) and 5 chronic graft-versus-host disease (cGVHD) after allo-HSCT. Three patients died of primary disease and 1bleeding in auto-HSCT group. One patient died of primary disease and 2 transplantation-related mortality in allo-HSCT group. The 3-year cumulative overall survival (OS) were 56% (95%CI 32%-100%) and 57% (95%CI 30%-100%) for auto-HSCT and allo-HSCT, respectively (P=0.979) . The 3-year cumulative progression-free survival (PFS) were 34% (95%CI 14%-85%) and 57% (95%CI 30%-100%) for auto-HSCT and allo-HSCT, respectively (P=0.451) . Conclusion: Both auto-HSCT and allo-HSCT were optimal choices for AITL. In clinical practice, which HSCT was better for AITL patients should be based on comprehensive factors including sensitivity to chemotherapy, risk stratification and disease status at transplantation.

Prevalence of pre-existing direct-acting antiviral agents resistance-associated variants in genotype 1 HCV/HIV co-infected patients / 上海交通大学学报（医学版）

Objective·To investigate the prevalence of pre-existing direct-acting antiviral agents (DAAs) resistance associated variants (RAVs) in genotype 1 hepatitis C virus (HCV)/human immunodeficiency virus (HIV) co-infected patients.Methods·All NS3 and NSSB HCV sequences in genotype 1 HCV/HIV co-infected patients were retrieved from NCBI GenBank database.And sequences were aligned and analyzed using software MEGA 5.0.Results·In total,the overall prevalence of DAAs RAVs in NS3 region was high (26.06％ and 38.18％,respectively),no matter in genotype 1a or genotype 1b.In genotype 1a,the high prevalence of RAVs mainly presented in the position Q80 (8.45％).In genotype lb,S122 RAV was most observed (36.36％).It is worth noting that,RAVs in NS5B region were rare observed (0.77％) in this study,especially as no RAV was detected in any sequence of genotype 1a patients.Conclusion·The prevalence of pre-existing RAVs is high in NS3 region but rare in NS5B region in HCV/HIV co-infected patients,suggesting that NS5B inhibitors based combination regions are a better choice for HCV/HIV co-infected patients.

Percutaneous dynamic release in stress position by acupotomy in treating severe scapulohumeral periarthritis / 中国骨伤

<p><b>OBJECTIVE</b>To investigate the clinical efficacy of acupotomy stress position percutaneous dynamic release for severe shoulder periarthritis.</p><p><b>METHODS</b>From April 2012 to August 2016, 160 patients with severe shoulder periarthritis were randomly divided into treatment group and control group. Among them, 80 patients in treatment group were treated with acupotomy stress position percutaneous dynamic release including 32 males and 48 females with an average of(52.47±9.04)years old ranging from 40 to 74 years old;the courses of disease was(20.72±9.55)months on average. The other 80 patients in control group were treated with simple joint loosening according to Maitland technique in grade III-IV therapy, once a day, 15 to 20 min each time, and 10 d for 1 course, for a total of 2 courses, including 33 males and 47 females with an average of (53.19±10.18) years old ranging from 42 to 75 years old; the average course of disease was (21.98 ±8.99) months. After operation, the shoulder muscles training and shoulder joint activity training were routinely conducted, the treatment lasted for 3 weeks. The visual analogue scale(VAS) and Constant-Murley shoulder function score were observed and compared between the two groups before treatment and 3 weeks, 3, 6 months after treatment.</p><p><b>RESULTS</b>The VAS scores of the treatment group at 3 weeks, 3 and 6 months after treatment were all lower than those of the control group(<0.05). The shoulder joint function Constant-Murley scores of the treatment group at 3 weeks, 3 and 6 months after treatment were higher than those of the control group (<0.05); the result was excellent in 59 cases, good in 18 cases, fair in 3 cases in the treatment group; excellent in 15 cases, good in 31 cases, fair in 23 cases, poor in 11 cases in the control group, and the difference between the two groups was statistically significant(<0.01).</p><p><b>CONCLUSIONS</b>Treatment of severe shoulder periarthritis with acupotomy stress position percutaneous dynamic release can obviously improve the shoulder joint function and pain, according to the different parts of the shoulder joint pain and function limitation, the corresponding shoulder stress and body position should be designed and maintained during the treatment process, and the angle of stress position gradually increased by loosening the adhesion, which is the key to ensure the curative effect.</p>

Reduced Macular Vascular Density in Myopic Eyes / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Morphological changes of the vasculature system in patients with myopia have been observed by Doppler ultrasound and fundus fluorescein angiography (FFA); however, these studies have limitations. Doppler ultrasound provides low-resolution images which are mainly obtained from visualized large vessels, and FFA is an invasive examination. Optic coherence tomography (OCT) angiography is a noninvasive, high-resolution measurement for vascular density. The purpose of this study was to investigate the change of vascular density in myopic eyes using OCT angiography.</p><p><b>METHODS</b>This cross-sectional study includes a total of 91 eyes from 47 participants including control, moderate, and high myopia that were evaluated by OCT angiography. Patients with myopia were recruited from the Refractive Department, Shenzhen Aier Eye Hospital, from August 5, 2015 to April 1, 2016. Emmetropic eyes were from healthy volunteers. The vascular density at macula and optic disc regions, ganglion cell complex (GCC) thickness, and retinal nerve fiber layer (RNFL) thickness were measured. Their relationships with axial length (AL) and refractive error were analyzed. One-way analysis of variance (ANOVA), Pearson's correlation, and generalized estimating equation were used for statistical analysis.</p><p><b>RESULTS</b>Both superficial and deep macular vascular density were highest in control (25.64% ± 3.76% and 37.12% ± 3.66%, respectively), then in moderate myopia (21.15% ± 5.33% and 35.35% ± 5.50%, respectively), and lowest in high myopia group (19.64% ± 3.87% and 32.81% ± 6.29%, respectively) (F = 13.74 and 4.57, respectively; both P < 0.001). Both superficial (β = -0.850 and 0.460, respectively) and deep (β = -0.766 and 0.396, respectively) macular vascular density were associated with AL and spherical equivalent (all P < 0.001). Superficial macular vascular density was associated with GCC thickness (β = 0.244, P = 0.040), independent of spherical equivalent. The vascular density in optic disc region had no difference among the three groups, and it was not associated with AL, spherical equivalent, or RNFL thickness.</p><p><b>CONCLUSION</b>Our results suggested that with the increase of myopia, the vascular density decreased in macular region, but not in optic disc region.</p>

Effect of miR-101 on biological characteristics of colorectal cancer cell line SW620 / 南方医科大学学报

<p><b>OBJECTIVE</b>To investigate the effect of miR-101 on biological behaviors of colorectal cancer cell line SW620.</p><p><b>METHODS</b>CCK-8 method, colony formation assay, cell cycle analysis and apoptosis analysis were applied to assess the effects of miR-101 on cell proliferation, invasion and apoptosis of SW620 cells.</p><p><b>RESULTS</b>Over-expression of miR-101 in SW620 cells significantly suppressed the cell proliferation and attenuated the colony-forming ability of the cells. Flow cytometry showed that over-expression of miR-101 in SW620 cells caused obvious cell cycle arrest in G2/M and 1/ phases, and significantly increased the cell apoptosis rate.</p><p><b>CONCLUSION</b>Over-expression of miR-101 can inhibit the proliferation, cause cell cycle arrest and promote apoptosis of colorectal cancer SW620 cells.</p>

MiR-135b promotes proliferation of endometrial carcinoma cells by targeting FOXO1 / 南方医科大学学报

<p><b>OBJECTIVE</b>To explore the expression of miR-135b in endometrial carcinoma and the mechanism by which miR-135b promotes the proliferation of endometrial cancer cells.</p><p><b>METHODS</b>The expressions of miR-135b and FOXO1 were using RT-PCR detected in 22 fresh endometrial cancer tissues and paired adjacent tissues and also in endometrial cancer cell lines JEC, Ishikawa, HEC-1-B, and RL-952. The RL-952 and Ishikawa cell lines were transfected with miR-135b mimics or inhibitors, and the changes in their proliferative activity were detected with MTT assay; the expressions of FOXO1 mRNA and protein were detected by RT-PCR and Western blotting, respectively.</p><p><b>RESULTS</b>The expression of miRNA135b was significantly up-regulated and FOXO1 expression was down-regulated in endometrial carcinoma tissues as compared with the adjacent tissues (P<0.05). The mRNA expression of miR-135b was negatively correlated with the expression of FOXO1 in endometrial carcinoma. In RL-952 and Ishikawa cell lines, transfection with miR-135b mimics obviously promoted the cell proliferation (P<0.05). Up-regulation of miR-135b significantly decreased the expressions of FOXO1 protein and mRNA (P<0.05), and down- regulation of miR-135b increased FOXO1 expressions (P<0.05).</p><p><b>CONCLUSIONS</b>MiR-135b plays an important role in the occurrence and development of endometrial carcinoma partially by regulating its target gene FOXO1.</p>

Relationship between subfoveal choroidal thickness and both choroidal hemodynamic index and glycosylated hemoglobin in diabetic subjects / 国际眼科杂志(Guoji Yanke Zazhi)

AlM: To investigate the relationship between the subfoveal choroidal thickness ( SFCT) and both choroidal hemodynamic index and glycosylated hemoglobin in diabetic subjects.METHODS:Seventy-eight type 2 diabetic patients (156 eyes) from ophthalmology and endocrinology ward of our hospital were enrolled in this study, including 39 females and 39 males, with a mean age of (59. 8±6. 2)years. According to early treatment diabetic retinopathy study ( ETDRS) grading method, all samples were divided into diabetic retinopathy ( DR ) group, mild or moderate nonproliferative diabetic retinopathy group, severe nonproliferative diabetic retinopathy ( NPDR) group and proliferative diabetic retinopathy ( PDR ) group. The SFCT and choroidal hemodynamic index were measured by enhanced depth imaging optical coherence tomography ( EDl-OCT ) and Color Doppler lmaging. Recording glycosylated hemoglobin content of all samples. Using multivariate linear regression to analyse the relationship between the SFCT and both choroidal hemodynamic index and glycosylated hemoglobin.RESULTS: The end diastolic velocity ( EDV ) was significant higher and the SFCT was significant thinner in no diabetic retinopathy ( NDR) group than other groups. There was no significant difference of peak systolic velocity ( PSV ) between four groups. The resistance index ( Rl) was significant higher in severe NPDR group than NDR group and mild or moderate group, the Rl in PDR group was hihgest than other group with statistically significance. The SFCT was correlated positively ( b =0. 540,P<0. 001) with the glycosylated hemoglobin. No significant correlation was found between the SFCT and the choroidal hemodynamic index (DR,P=0. 341;PSV,P=0. 770;EDV,P=0. 131;Rl,P=0. 084).CONCLUSlON: Our results suggest that there is no significant correlations between the SFCT and the choroidal hemodynamic index; glycosylated hemoglobin is one of the factors that affect the SFCT in diabetic patients.

Expression of Hydrodynamic Injection-mediated PD-L1 in Myeloablative Conditioning Mouse Model / 中国实验血液学杂志

<p><b>OBJECTIVE</b>To establish the mouse model for the expression of PD-L1 by hydrodynamic injection and to study the effects of myeloablative conditioning on hydrodynamic injection-mediated PD-L1 expression.</p><p><b>METHODS</b>Plasmid amplification, hydrodynamic injection, collagenase perfusion, real time PCR, ELISA and flow cytometry were applied to test the expression and function of PD-L1. Also, animal models were set up to test the effects of chemical or radiactive myeloablative conditioning on hydrodynamic injection-mediated PD-L1 expression.</p><p><b>RESULTS</b>The expression of PD-L1 mRNA and protein could be detected as early as 8 h after hyrodynamic injection and reached peak expression by 24 h, and returned to baseline level by 7 d after injection. Serum PD-L1 level reached to 100 µg/ml as early as 24 h after injection and plateaued at 7 d after injection. Serum PD-L1 persisted for 3 weeks and declined to baseline after 1 month of hydrodynamic injection. The PD-L1 function induced by hydrodynamic injection was consistent with literature reports. At each time point, the PD-L1 expression was not different significantly between the myeloablative conditioning group and control group; the mice transfected with PD-L1 showed a higher survival rate than that in control group.</p><p><b>CONCLUSION</b>Myeloablative conditioning does not affect hydrodynamic injection-mediated PD-L1 expression, indicating that the PD-L1 can be used in HSCT mouse model.</p>

Numerical simulation of pulse wave propagation and parameter analysis based on transmission line model / 医用生物力学

Objective To numerically simulate the propagation of pulse wave in human arterial tree by proposing a novel calculation method which combines a transmission line model and a recursive algorithm of input impedance, and to study the effects of individual differences and arterial tree parameters on pulse wave so as to provide references for the analysis on physiologic and pathologic characteristics of human arterial tree. MethodsThe transmission line model of human arterial tree was constructed, which consisted of 55-segment large and medium sized arteries. The recursive algorithm was applied to compute the input impedance of arterial tree at each point. The blood pressures and flows of 55 arteries were calculated and showed in the distribution graphs. Based on this method, the effects of height, heart rate, stroke volume, internal radius and wall thickness on pulse wave propagation and blood pressure distribution were compared. Results The simulation results were in good agreement with the general rules of pulse wave propagation. The propagation of pulse wave in arterial tree showed significantly different characteristics for different parameters. Conclusions The proposed method can effectively simulate the propagation of pulse wave in arterial tree and accurately reflect the effects of individual differences and hemodynamics parameters on pulse wave propagation, and it is an important assistant means for the pathophysiologic analysis and diagnosis of human arterial tree.

Construction of has-miR-335 lentiviral vector and verification of the target gene of miR-335 / 南方医科大学学报

<p><b>OBJECTIVE</b>To construct a lentiviral vector of miR-335 gene and verify the target gene of miR-335.</p><p><b>METHODS</b>The precursor sequence of miR-335 gene was amplified from the genomic DNA by PCR and cloned into the lentiviral vector PLVTHM labeled with GFP. Real-time quantitative RT-PCR was used to detect miR-335 and RASA1 expression in different colorectal cancer cell lines. A recombinant vector psiCHECK-2-RASA1 containing RASA1 3'UTR was constructed followed by site-directed mutagenesis of RASA1 3'UTR to establish the vector psiCHECK-2-RASA1-Mut. Co-transfection of hsa-mir-335 or a NC with these recombined vectors in HEK293A and SW480 cells was performed, and dual-luciferase reporter assay was utilized to examine the changes in luciferase activities. The recombinant PLVTHM-miR335 plasmid was packaged into mature lentivirus by 293FT cells and used to infect SW620 cells. Flow cytometry was employed for sorting the GFP+ cells. The expression of miR-335 and RASA1 were determined by qRT-PCR, and Western blotting was used to detect the expression of RASA1 protein in SW620 cell lines.</p><p><b>RESULTS</b>The recombinant lentiviral vector PLVTHM-miR335, psiCHECK-2-RASA1 and the mutation expression vector psiCHECK-2-RASA1-Mut were successfully constructed. Dual-luciferase reporter assay showed that miR-335 decreased luciferase activity in cells co-transfected with psiCHECK-2-RASA1. The expression of miR-335 in SW620 cells infected with the lentivirus PLVTHM-miR335 was significantly increased, but the expression of RASA1 showed only slight changes. Overexpression of miR-335 suppressed the expression of RASA1 protein in SW620 cells.</p><p><b>CONCLUSION</b>We have successfully constructed the lentiviral vector containing mir-335 gene and a SW620 cell line with miR-335 overexpression. MiR-335 can suppress RASA1 gene expression by targeting the specific sequence of RASA1 3'UTR.</p>

Long-term culture and identification of CD133⁺ hematopoietic progenitor cells from human umbilical cord blood / 南方医科大学学报

<p><b>OBJECTIVE</b>To isolate CD133(+) hematopoietic progenitor cells from human umbilical cord blood and optimize the culture condition for maintaining their stem cell characteristics.</p><p><b>METHODS</b>CD133(+) hematopoietic progenitor cells were isolated from human umbilical cord blood using magnetic cell sorting system, and the cells were detected by flow cytometry. Four methods were used for culturing cells. After 8 weeks' culture, cytomorphology, flow cytometry, immunocytochemistry and immunofluorescence assay were used to identify the characteristics of the stem cells.</p><p><b>RESULTS</b>Over 80% of CD133(+) hematopoietic progenitor cells were isolated from human umbilical cord blood using magnetic cell sorting system. The cells were effectively expanded using optimized serum-free medium after 8 weeks of cell culture, whereas the cells in other media differentiated into adherent cells in a poor state.</p><p><b>CONCLUSION</b>The optimized serum-free medium allows effective expansion of CD133(+) hematopoietic progenitor cells that maintain stem cell characteristics after a long-term culture.</p>

Touchdown PCR and overlap extension PCR for generating CD133(+) cancer stem cell-selective adenovirus vector / 南方医科大学学报

<p><b>OBJECTIVE</b>To construct a replication-incompetent adenovirus vector targeting cancer stem cells by modified touchdown PCR and overlap extension PCR and investigate its infection efficiency in CD133(+) SW480 cells in vitro.</p><p><b>METHODS</b>The two portions of the fiber gene encoding the Ad5 fiber knob domain with the HI loop deleted were amplified using two pairs of designed primers and then linked by overlap extension PCR. The product obtained was identified by sequencing and inserted into prokaryotic expression vector pEGFP-N1. The product, pEGFP-N1 KNOBδHI, contained a unique EcoRV restriction site in the deleted portion of the sequence encoding the HI loop. The gene sequences of the adenovirus fiber were amplified using both common PCR and overlap extension PCR, then identified by sequencing and inserted into pNEB193, resulting in pNEB-F5. CD133(+) SW480 cells were infected with the generated adenovirus vectors Ad5-GFP and Ad5FHI-GFP to investigate the infection efficiency using fluorescent microscope.</p><p><b>RESULTS</b>The target fragments of expected sizes were amplified by touchdown PCR and overlap extension PCR, but not by common PCR. Ad5FHI-GFP showed a higher infection efficiency than Ad5-GFP in CD133(+) SW480 cells.</p><p><b>CONCLUSION</b>Compared with common PCR, touchdown PCR and overlap extension PCR can significantly improve the specificity and efficiency of the PCR products for constructing CD133(+) cancer stem cell-selective adenovirus type 5 vector, which provides carriers for tumor-targeted gene therapy.</p>

Selection and identification of specific-binding peptides for cancer stem cell surface marker CD133 / 南方医科大学学报

<p><b>OBJECTIVE</b>To select the peptides that specifically bind human cancer stem cell surface marker CD133 from the Ph.D.-7>(TM) phage peptide library.</p><p><b>METHODS</b>With a biotinylated extracellular fragment of human cancer stem cell surface marker CD133 as the target protein, the CD133 high-affinity peptides were screened from the phage peptide library by liquid phase panning. The clones with high-binding force with human CD133 were then identified by sandwich ELISA and their single-stranded DNA was extracted to test the specificity by competitive ELISA. The amino acid sequences of the selected peptides derived from the phage DNA sequences were synthesized after sequence alignment analysis, and their capacity of binding with colorectal carcinoma cells was assessed by immunofluorescence technique.</p><p><b>RESULTS</b>After 4 rounds of liquid phase selection, the phages capable of specific binding with human CD133 were effectively enriched, with an enrichment ratio of 388 times compared to that at the fourth and first rounds. Thirteen out of the 20 clones from the fourth round of panning were identified as positive clones, among which 11 had identical amino acid sequence of TISWPPR, and 2 had the sequence of STTKLAL, and the former sequence showed a stronger binding specificity to CD133.</p><p><b>CONCLUSION</b>We have successfully obtained a peptide that specifically binds human CD133 from the Ph.D.-7(TM) phage peptide library, demonstrating the feasibility of screening small molecule high-affinity polypeptides from phage peptide library by liquid-phase panning.</p>

Recursive calculation and parameter analysis on input impedance of arterial tree based on electric network model / 医用生物力学

Objective In order to provide a reference for analysis of the pathophysiological changes of human arterial tree, the effects of the arterial parameters on input impedance of arterial tree were studied by proposing a recursive algorithm to calculate the input impedance. Method A distributed electric network model was constructed for the arterial tree which consisted of 55 segments accounting for all the central vessels and major peripheral arteries. By setting up the peripheral resistance of the model and constructing a unilateral data link list of the arterial tree, the recursive algorithm was employed to compute the input impedance of ascending aorta in the arterial tree. Then, comparisons of the effects of arterial compliance, peripheral resistance, length, internal radius and wall thickness on the input impedance were made based on the recursive algorithm. Results The computational results were in accord with experimental data and results of other models, and proved the validity of the model and algorithm. The input impedances of the arterial tree showed significant influence with different characteristics for different parameters. Conclusions The input impedance of arterial tree can reflect the changes of the hemodynamic parameters effectively, which is an important reference for the pathophysiological diagnosis of human arterial tree.

The effect of oral alpha-lipoic acid in overweight/obese individuals on brachial-ankle pulse wave velocity and supine blood pressure:a randomized,crossover,double-blind,placebo-controlled trial / 中华流行病学杂志

Objective To investigate the effect of oral alpha-lipoic acid (ALA) supplement on brachial-ankle pulse wave velocity (baPWV),supine systolic blood pressure (SBP) and diastolic blood pressure (DBP) in overweight/obese individuals.An 8-week double-blind,randomized,placebo-controlled and cross-over trial with a 4-week washout between cross-over periods.Methods Sixty-three males and 40 females aged 22-57 years old who met the inclusion criteria as (1) Han ethnicity;(2) 20-60 years old;(3) BMI≥25 kg/m2 and having at least one of the following risk factors:borderline hypertension (130 mm Hg≤SBP＜140 mm Hg and/or 85 mm Hg≤supine DBP＜90 mm Hg),dyslipidemia(fasting total cholesterol≥5.2 mmol/L or HDL-C＜1.04 mmol/L),or impaired fasting glucose (6.1 mmol/L≤fasting glucose＜7.0 mmol/L);(4)Not on any antioxidant gender.Group 1 received 8 weeks ALA (1200 mg/day) followed by 4-week washout period and followed by another 8 weeks placebo;while Group 2 received 8 weeks placebo (1200 mg/day)followed by 4-week washout period,and followed by ALA treatment for 8 weeks.BaPWV and supine blood pressure were measured at the beginning of 1st phase and 2nd phase and at the endpoint of the whole trial.Mixed effect linear regression model was performed to compare the change of baPWV and supine blood pressure between ALA group and placebo group.Results BaPWV decreased -33.03 cm/s ± 130.70 cm/s for ALA group and increased 5.66 cm/s ± 139.89 cm/s for placebo group,supine systolic blood pressure decreased -4.09 mm Hg±9.18 mm Hg for ALA group and -2.32 mm Hg±8.16 mm Hg for placebo group.Supine diastolic blood pressure decreased -1.29 mm Hg ± 6.55 mm Hg for ALA group and -0.48 mm Hg±6.63 mm Hg for placebo group.These three mix-effect models did not show significant effect of ALA treatment after adjustment on baseline values,sex,age,treatment sequence or period.Conclusion The current trial did not provide evidence that oral intake of ALA for 8 weeks had significant effects on lowering baPWV,supine systolic blood pressure or supine diastolic blood pressure.