AIM:To investigate the effect of calcitonin gene-related peptide (CGRP)transfection into c-kitpos cardiac stem cells (c-kit+CSCs) on the cell viability.METHODS: Under the sterile condition, the auricles of SD rats were taken out , and then c-kit +CSCs were collected through enzyme digestion and immunomagnetic bead separation (MACS).The cells were identified by flow cytometry .c-kit +CSCs were transfected with enhanced green fluorescent pro-tein CGRP lentiviral vector ( Lv-EGFP-CGRP) or enhanced green fluorescent protein lentiviral vector ( Lv-EGFP).The cells were randomly divided into Lv-EGFP-CGRP-CSCs group , Lv-EGFP-CSCs group and CSCs group .The transfection was observed under the fluorescence microscope .The transfection efficiency was detected by flow cytometry .The CGRP protein secretion in the cell culture supernatants was detected by ELISA .The viability of c-kit+CSCs transfected with Lv-EGFP-CGRP or Lv-EGFP was measured by CCK-8 assay.RESULTS:c-kit+CSCs were isolated and cultured successfully .The expression positive rate of c-kit was 91.0%and the expression positive rates of CD 45 and CD34 were 4.5% and 4.0%, respectively.After transfected with lentivirus for 48 h, the stable fluorescence in c-kit +CSCs was observed under fluores-cence microscope .The transfection efficiency were 80%when MOI was 20.The level of CGRP was significantly increased in Lv-ECFP-CGRP-CSCs group compared with Lv-EGFP-CSCs group and CSCs group (P<0.05).Meanwhile, transfection with lentiviral vector in each group did not affect the viability of c-kit+CSCs.CONCLUSION:Transfection of Lv-EGFP-CGRP into c-kit+CSCs was successful .The secretion of CGRP was found in the transfected c-kit+CSCs and the viability was not changed after transfection .CGRP-modified c-kit+CSCs may play a role in treating myocardial infarction .

Carbon Monoxide ; physiology ; Humans ; Nervous System Diseases ; physiopathology ; Neurotransmitter Agents ; physiology ; Nitric Oxide ; physiology

Adolescent ; Anticonvulsants ; therapeutic use ; Body Mass Index ; Child ; Child, Preschool ; Epilepsy ; blood ; drug therapy ; Female ; Galanin ; blood ; Humans ; Infant ; Male ; Valproic Acid ; blood ; therapeutic use

Objective This study aims to explore the application value of tuberculosis T lymphocytes enzyme-linked immune SPOT test (T-SPOT.TB) on early diagnosis of tuberculosis.Methods The TB infection in 189 inpatients suspected tuberculosis in pneumology department of Shaanxi Provincial People's Hospital was detected with T-SPOT.TB,fluorescence RQPCR,tuberculosis (TB-Ab)protein chip and PPD methods.Results The sensitivity of four methods was 91.54％ (119/130),73.85％(96/130),63.08％(82/130) and 57.69％ (75/130) respectively and the specificity of those was 89.83％ (53/59),86.44％(51/59),67.79％(40/59) and 66.10％(39/59),respectively.The sensitivity of T-SPOT.TB method was statistically higher than those of other three tests,respectively (P＜0.05).The specificity of T-SPOT.TB was significantly higher than those of TB-AB and PPD (P＜0.05),but there was no statistical difference between RQ-PCR and T-SPOT.TB (P＞0.05).The positive predictive values of T-SPOT.TB,fluorescent quantitative PCR,TB-Ab and PPD assays were 95.2％ (119/1250),92.3％ (96/104),81.2％ (82/101) and 78.9％ (75/95) respectively while the negative predictive values of those were 82.8％ (53/64),60％ (51/85),45.5％ (40/88) and 41.5％ (39/94),respectively.The false-positive rates (misdiagnosis rate) of four assays were 10.2％ (6/59),13.6％ (8/59),32.2％ (19/59) and 33.9％ (20/59) respectively and the false-negative rates (rates of missed diagnosis) of those were 8.5％ (11/130),26.2％ (34/130),36.9％ (48/130)and 42.3 ％ (55/130),respectively.The negative likelihood ratios of T SPOT.TB,fluorescent quantitative PCR,TB-Ab and PPD assays were 0.11,0.16,0.48 and 0.51 respectively,meanwhile the positive likelihood ratios of T-SPOT.TB,fluorescent quantitative PCR,TB-Ab andPPD assays were 9.0,5.4,2.0 and 1.7,respectively.What' s more,the diagnostic accordance rates of the four assays were 91.0％ (172 189),77.8％ (147 189).64.6％ (122/189) and 60.3％ (114/189),respectively.Conclusion T-SPOT.TB test is a more sensitive and specific method and of great significance to the early diagnosis of TB,which has more clinical value in different stages of tuberculosis diagnosis.

AIM: To investigate hemodynamic alterations of retrobulbar vessels in proliferative diabetic retinopathy ( PDR) patients with anterior segment neovascularization, before and 3mo after vitrectomy combined with panretinal photocoagulation and to explore the clinical significance.
●METHODS: Color Doppler flow imaging ( CDFl ) was used for measurement of blood flow velocities and resistive indexes ( Rl ) of the ophthalmic artery ( OA ) , short posterior ciliary arteries ( sPCA ) and central retinal artery ( CRA ) in 21 eyes of 21 PDR patients with anterior segment neovascularization. CDFl parameters were obtained before and 3mo after vitrectomy combined with panretinal photocoagulation ( PRP) .
● RESULTS: Peak systolic velocity ( PSV ) and end diastolic velocity ( EVD ) of CRA were significantly increased after surgeries, Rl were decreased significantly (P<0. 05). Parameters of sPCA and OA have no change after surgeries (P>0. 05).
●CONCLUSION: Vitrectomy combined with panretinal photocoagulation might increase the velocity of CRA, decrease Rl and improve ocular blood supply postoperatively. lt may delay or prevent the process of neovascular glaucoma.

Objective To evaluate the the effect of microalbuminuria of combined treatment with fosinopril and losartan,or fosinopril,losartan monotherapy in patients with hypertension.Methods In this double-blind, intention to treat study,136 patients with hypertension were randomly assigned to receive fosinopril 10 mg/d(n= 50),losartan 50 mg/d(n=41),or a combination of fosinopril 5 mg and losartan 25 mg (n=45) qd for 4 weeks, followed by titrating to the maximum recommended doses for another 4 weeks.The primary endpoint was the difference of mean sitting blood pressure and microalbuminuria excretion at baseline and week 8.Results At week 8,the combination of fosinopril and losartan therapy lowered mean mieroalbuminuria from baseline by 26.1?10~(-8) mol/L,significantly more than either monotherapy approaches (fosinopril 20 mg,18.3?10~(-8)mol/L,P

Objective: To explore the diagnosis, treatment and prognosis of autoimmune hemolytic anemia (AIHA) after allo-HSCT in patients with thalassemia major (TM). Methods: A retrospective analysis of AIHA status after allo-HSCT in 291 TM patients from July 2007 to December 2017 was conducted. Results: Five of the 291 TM patients (1.72%) were diagnosed with post-transplant AIHA. The median time of AIHA was 7 (5-12) months after HSCT. All post-transplant AIHA patients were positive in direct and indirect Coombs test, the main clinical manifestations were dizziness, fatigue, pale complexion, skin and sclera yellow, and soy sauce urine. The incidence of AIHA was higher after unrelated donor transplantation (6.36%, 4/63) compared with that of sibling donor transplantation (0.43%, 1/228). One patient who received only prednison was dead. Four patients who received rituximab combined with prednisolone were alive, Coombs test in two of them were negative. Conclusions: AIHA after allo-HSCT developed in 1.72% patients with TM. Monitoring of Coombs test was important for diagnosis of post-transplant AIHA. The incidence of post-transplant AIHA was higher in unrelated donors compared with that of sibling donors transplantation. Treatment of rituximab combined glucocorticoid was effective strategy for post-transplant AIHA.
Anemia, Hemolytic, Autoimmune ; Coombs Test ; Hematopoietic Stem Cell Transplantation ; Humans ; Retrospective Studies ; beta-Thalassemia

OBJECTIVETo explore the quality of the embryos, clinical outcomes and birth defects resulting from intracytoplasmic sperm injection (ICSI) treatment using sperms of different origins and parameters.

METHODSA total of 980 ICSI-ET (embryo-transfer) cycles were divided into 4 groups, namely normal sperm or mild oligozoospermia group (group A), severe oligozoospermia group (group B), epididymal aspirates group (group C) and testicular biopsies group (group D). The cleavage rate, embryo quality, fertilization rate, clinical pregnancy rate, and rates of birth defects were compared between the groups.

RESULTSThe fertilization rate, cleavage rate and good-quality embryo rate were not significantly different among the 4 groups (P>0.05), and the embryo implantation rate and clinical pregnancy rate were significantly higher in group C than in groups A and B (P<0.05). Groups A, B and C showed no significant differences in the rates of birth defects (P>0.05), and no birth defects occurred in group D.

CONCLUSIONSSperms of different parameters and origins used in ICSI treatment can achieve similar fertilization rate, good-quality embryo rate and delivery rate. The embryo implantation rate and clinical pregnancy rate of epididymal sperm group are higher than those of ejaculated groups, possibly due to the younger age of the patients and a greater number of oocytes retrieved in group C than in groups A and B.

Congenital Abnormalities ; etiology ; Female ; Fertilization ; Humans ; Male ; Sperm Injections, Intracytoplasmic ; adverse effects ; Spermatozoa ; Treatment Outcome

OBJECTIVETo explore the effect of Chinese herbs (CH) for cool-moistening and freeing collaterals on gastro-dynamic disturbance in patients of diabetes mellitus type 2 with gastroparesis (DM-GP).

METHODSFifty-three patients of DM-GP were enrolled and treated with CH (n = 28) and Cisapride (n = 25) respectively for 4 weeks, the changes of gastrin and electro-gastrogram (EGG) before and after treatment were observed.

RESULTSAfter treatment, the EGG improved significantly, showing the rhythm significantly improved, and level of serum gastrin lowered significantly, as compared with those before treatment, the difference was significant (P<0.01), but insignificant difference was found between the two groups. Fifteen patients in each group were followed-up afar stopping medication for 3 months, recurrence occurred in 1 patient of CH treated group, and 2 patients of Cisapride treated group. No adverse reaction was found in the rest patients.

CONCLUSIONCH could obviously improve the gastro-intestinal motility and hormones abnormality.

Adult ; Aged ; Diabetes Mellitus, Type 2 ; blood ; complications ; drug therapy ; Drugs, Chinese Herbal ; therapeutic use ; Electromyography ; Electrophysiology ; Female ; Gastric Emptying ; drug effects ; Gastrins ; blood ; Gastrointestinal Agents ; therapeutic use ; Gastrointestinal Motility ; drug effects ; Gastroparesis ; drug therapy ; etiology ; physiopathology ; Humans ; Male ; Middle Aged ; Phytotherapy

Objective To investigate the epidemiology of viral infection in elderly patients to contract acute exacerbations of chronic obstructive pulmonary disease (AECOPD)in Minhang district of Shanghai from 2010 to 2012,and to study the relationships between viral infection and clinical features.Methods The elderly patients (age >70 year old)with AECOPD admitted from September 2010 to November 2012 were enrolled for study.The patients who couldn't complete lung function test were excluded.The pharyngeal swabs (PS)were taken from each patient within the first 24 h after admission.Nine respiratory viruses and their subtypes from pharyngeal swabs were detected by the nested multiplex polymerase chain reaction (PCR)method,including influenza virus A (FluA),2009 influenza A (H1N1 )virus (09FluH1 ), influenza virus B (FluB),respiratory syncytial virus A (RSVA)and B (RSVB),human coronavirus-229E (hCOV-229E),human coronavirus-NL63 (hCOV-NL63 ),human coronavirus-OC43 (hCOV-OC43 ), human coronavirus-HKU1 (hCOV-HKU1),human parainfluenza virus 1-4 (hPIV1-4),human adenovirus (hAdV),human boca virus (hBoV),human metapneumo-virus (hMPV)and human rhinovirus (hRV). According to the PCR results,all patients were divided into positive viral infection group and negative viral infection group.The relationships between viral infection and clinical features were analyzed.Results Sixty patients were eligible for study.Of them,14 patients were found to be positive for virus infection including a triple infected patient (FluB,hRV and hROV)and 46 patients were negative for virus infection.The viral pathogens detected in the positive viral group were:9 cases of hRV (15.00%),2 cases of hPIV (3.33%),2 cases of hCOV (3.33%),2 cases of FluB (3.33%)and 1 cases of RSV (1.67%).The mortality in the positive viral group was higher than that in the negative viral group.However,the other clinical characteristics between the two groups had no significant differences. Conclusions Human rhinovirus was the most common viral pathogen in elderly patients with AECOPD.Viral infection might be associated with the prognosis.However,the patients with viral infection are lack of specific clinical characteristics,therefore,the prompt diagnosis before careful study would be difficult.