Objective To investigate the tumor-inhibitory effect of cytokine-induced killer cells(CIK)co-cul-tured with dendritic cells (DC)pulsed by breast cancer stem cell antigen on the same tumor-bearing mice. Methods Breast cancer stem cells were isolated from the cell line of MCF-7/ADR and extract lyses antigen of the stem cell was saved. DC and CIK derived from peripheral blood mononuclear cells of healthy individuals were co-cultured and pulsed or un-pulsed by the above antigen lyses. This DC+CIK were injected to breast tumorbearing mice (BCSC-AP-DC+CIK group), and were used to compared with the common breast cancer cell antigen (rather than breast cancer stem cell antigen) pulsed DC+CIK group(AP-DC+CIK group), DC+CIK group, CIK CIK group and normal saline group(NS group). The tumor-inhibitory effect were evaluated and compared among all 5 groups through the tumor size, TdT-mediated dUTP nick end labeling test (TUNEL), examining expression level of bcl-2 and bax by immunohistochemistry. Results The tumor size in each group before and after therapy and the tumor size after therapy between each group was of significant difference(P＜0.05). The maximum size is NS group(3.625±0.093)cm3 and BCSC-AP-DC+CIK group is minimum,which is (1.234±0.131)cm3. BC-SC-AP-DC+CIK group is of highest expression of bax and apoptotic index value, lowest bcl-2 expression in all 5 groups. Conclusion The CIK co-cultured with DC pulsed breast cancer stem cell antigen was more effective to induce apoptosis of breast cancer cells than those of CIK cells co-cultured with DC pulsed breast cancer cell antigen,CIK cells co-cultured with DC and CIK cells.

Objective To investigate the relationship between the single nucleotide polymorphism (SNP)of KCNJ1 1 (inwardly rectifying potassium channel,subfamily J,member 1 1)gene and essential hypertension (EH)in Xinjiang Kazak population.Methods The polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP ) method was used to detect genotypes of rs2285676 polymorphism of KCNJ1 1, including 126 hypertensives (EH group)and 126 normotensives (NT group)in Xinjiang Kazak population.Multiple-factor unconditional Logistic regression analysis was used to evaluate the risk factors for hypertension in Xinjiang Kazak. Results Logistic regression analysis showed that KCNJ1 1-rs2285676 genotypes,sex,weight,total cholesterol and triglyceride were not associated with hypertension;body mass index (BMI)and high-density lipoprotein (HDL) were protective factors affecting hypertension while low-density lipoprotein (LDL ) was a risk factor for hypertension.Genotypes (TT,CT and CC)and allele frequencies (T and C)of the SNP of rs2285676 in KCNJ1 1 gene were 50.00%,48.41%,1.59%,74.21% and 25.79% in EH group,respectively.The differences between EH and normal control were not significant (P >0.05).Conclusion The rs2285676 polymorphism of KCNJ1 1 is not associated with EH in Xinjiang Kazak population.

Objective Toinvestigatetheentropyoflocalfieldpotentials(LFPs)recordedinratmedialprefrontal cortex during a Y-maze working memory (WM) task, to provide computing support for neural coding mechanism.Methods Sixteen-channel LFPs were recorded from SD rats while they performed a Y-maze WM task.The data came from 4 rats, 20 trials (10 correct trials and 10 incorrect trials) per rat provided by laboratory of neurobiology in medicine,Tianjin Medical University.Original LFPs were preprocessed to remove 50 Hz power line noise and baseline drift.Multi-taper Fourier transform was applied to calculate spatial distributions of LFPs and band pass filter were used to extract characteristic signal.The entroy coding of 16 channel LFPs was as follows: the physiological window was set to be 500 ms, the step length of physiological window was set to be 125 ms, windows were added to LFPs data, and then LFPs entropy of each sliding window was computed and averaged to get the trend of multichannel entropy values duringthe WM task.Results The power of θ band (4-12 Hz) in LFPs increased.The averaged entropy value ofmultichannel θ band LFPs in correct trials was 0.939±-0.020, which were larger than those in the resting state, 0.795±0.031 (P＜0.05).Those during wrong WM task had no significant difference, which didn't encode the WM task.Conclusions The principal frequency band related to WM is the θ band and LFPs entropy encodes the WM effectively.

Objective To explore the relationship between KCNJ11-E23K polymorphism and essential hypertension (EH) in Kazakh from Xinjiang. Methods PCR-RFLP method was used to test KCNJ11-E23K genotypes of Kazakh from Xinjiang,including 237 EH patients and 221 normotension (NT). Logistic regression analysis was used to analyze the risk factors associated with EH. Results The frequencies of KCNJ11-E23K genotype (EE and (EK + KK)) and allele (E and K) were 34.18%, 65.82%, 61.60% and 38.40%respectively in EH group. There was a significant difference between two groups (P < 0.05). Weight and EE genotype were risk factors affecting EH in Kazakh from Xinjiang. Individual who carried EE genotype and allele E were 2.501 and 1.388 times than (EK + KK) and allele K suffered from EH respectively. Conclusion KCNJ11-E23K polymorphism was associated with EH in Kazakh from Xinjiang.

Aim To explore the differences in hyper-trophic marker genes such as atrial natriuretic peptide ( ANP) , brain natriuretic peptide ( BNP) and β-myo-sin heavy chain (β-MHC) genes in different models of cardiac hypertrophy. Methods Respectively using re-nal abdominal aortic coarctation ( AAC) , arteriovenous fistula ( AVF) and isoproterenol ( ISO) methods to es-tablish C57BL/6 mouse model of cardiac hypertrophy. After modeling, each mouse ’ s body weight ( BW ) , heart weight ( HW) and left ventricular weight ( LVW) were weighed, and the heart weight ( HW/BW) and left ventricular index ( LVW/BW ) were calculated;myocardium by HE staining, pathological morphologi-cal changes were observed; myocardium by immuno-histochemistry, ANP, BNP and β-MHC protein ex-pression was observed;myocardium by Real-time PCR detection, ANP, BNP and β-MHC mRNA expression was observed. Results Compared with control group, HW/BW and LVW/BW were increased in three mod-els. Through the light microscope, each mouse model showed varying degrees of cardiac hypertrophy. ANP, BNP and β-MHC were increased in the protein and mRNA expression. Compared with AAC group, AVF and ISO groups’ myocardial tissue ANP, BNP and β-MHC expression were decreased in the protein and mRNA expression. Conclusions Three cardiac hy-pertrophy models are successful. Cardiac tissue ANP, BNP and β-MHC expression in AAC model exceeds AVF and ISO model.

Objective To investigate the association of the Cx37 polymorphism of connexin gene with essential hyper?tension (EHT) in Xinjiang Han and Kazak population. Methods In Xinjiang region, 500 EHT patients (EHT group) were in?cluded in this study including Kazak 250 cases and Han 250 cases. Five hundred healthy volunteers (NT) were used as NT group including Kazak 250 cases and Han 250 cases. The values of age, body mass index (BMI), systolic blood pressure (SBP) and diastolic blood pressure (DBP), and other general clinical features were compared between two groups. The poly?morphism of Cx37 gene rs1630310, rs697372 and rs705193 SNP were compared between EHT and NT groups in the two eth?nic groups. Hardy-Weinberg equilibrium was used to detect the representation, and differences of genotype frequencies and gene frequency were calculated in two groups of Kazak and Han groups. Results There were significant differences in BMI, SBP, DBP, apolipoprotein ratios and homocysteine between EHT group and NT group in Kazak and Han groups (P<0.05). There were no significant differences in genotype frequencies and gene frequencies of rs705193 between EHT and NT groups (P>0.05). The differences of Kazak rs1630310 genotype and gene frequency were statistically significant (P<0.01). The frequency of Kazak rs697372 locus genotype was not significantly different (P>0.05), but the difference of gene frequen?cy was statistically significant (P<0.05). There were no significant differences in rs1630310 and rs697372 locus genotype and gene frequency in two groups of Han group. Conclusion Cx37 gene polymorphism is associated with the occurrence of EHT in Xinjiang Kazak population, which may be related with the rs1630310 and rs697372 polymorphism.

Objective To investigate the serum levels of 25-hydroxyvitamin D3 (25 (OH)D3) and urine vitamin D binding protein(uVDBP) in patients with diabetic nephropathy (DN),and to determine the relationship between 25 (OH) D3,uVDBP and DN,in order to provide a new method for early diagnosis and treatment of DN.Methods From January 2015 to December 2015,85 DN patients admitted into Weihai Municipal Hospital were selected.According to the ratio of UALB to UCR(UACR),the patients were divided into three groups.Type 2 diabetes had 28 cases of normal albuminuria group,31 cases of microalbuminuria group,and 26 cases of clinical albuminuria group.We also enrolled 25 healthy people who received outpatient service as control group.Serum 25 (OH) D3 levels were measured by chemiluminescence method.Urine VDBP levels were assayed by ELISA.FPG,HbA1 c,UREA,SCr,TC,TG were measured by electrochemiluminescence.Results The results showed that serum 25 (OH)D3 was significantly lower in the normal albuminuria group,microalbuminuria group and clinical proteinuria group than that in the control group (P ＜ 0.05),and there was statistically significant difference among the four groups [(20.04 ± 7.52) ng/mL,(16.54 ± 6.51) ng/mL,(10.77 ± 4.63) ng/mL,(29.65 ± 5.47) ng/mL,F =86.294,P ＜ 0.001].The results showed that uVDBP was significantly higher in the DN group than that in the control group(all P ＜ 0.05),and there was statistically significant difference among the four groups [(8.44 ± 3.20) mg/L,(14.22 ± 3.26) mg/L,(2 1.77 ± 5.87) mg/L,(4.95 ± 1.34) mg/L,F =125.583,P ＜ 0.001].Correlation analysis showed that serum 25 (OH) D3 decreased gradually with the increase of DN and negatively correlated with UACR (r =-0.575,P ＜ 0.01),while uVDBP level was positively correlated with UACR (r =0.436,P =0.015).Conclusion With the progress of DN,serum 25 (OH) D3 levels gradually decreased,indicating that 25 (OH) D3 may play an important role in the pathogenesis of DN;uVDBP may be an early diagnostic method for DN.

Objective:To investigate the effect of Mediterranean diet on blood glucose control and cardiovascular risk factors in patients with type 2 diabetes.Methods:As to December 2021, the PubMed, Cochrance Central Register of Controlled Trials and Cochrance Database, Cochranc Library, Embase, China National Knowledge Infrastructure and Wanfang Medical Network system were searched for clinical randomized controlled trials(RCTs) of Mediterranean diet in patients with type 2 diabetes to conduct Meta-analysis The main observation index were cardiovascular risk factors, and the mean difference and its 95% confidence interval were used to estimate the effect size.Results:There were six RCTs, and 1181 patients met the inclusion criteria and entered the Meta-analysis. Compared with the control group, the intervention group can significantly reduce the level of systolic blood pressure ( MD=-1.20, 95% CI-2.21 to -0.19) and diastolic blood pressure ( MD=-4.17, 95% CI-7.12 to -1.22) in patients with type 2 diabetes mellitus, but there were no significant difference in the level of TC ( MD=2.92, 95% CI-0.84 to-6.67), HDL ( MD=2.33, 95% CI-0.27 to -4.92) and LDL ( MD=-2.34, 95% CI-5.67 to -0.99) between the two groups (all P>0.05). Conclusions:The meta-analysis provided evidence the Mediterranean diet showed the beneficial improvements in blood pressure glycemic control, but the effect of Mediterranean diet on lipid profile was not significant, which needed further verification.

Objective:To investigate the effect of distilled water on the viability of human lens epithelial cells (LECs) cultured in vitro. Methods:A total of 156 anterior capsule specimens were collected from 156 patients (156 eyes) who were diagnosed with age-related cataract during phacoemulsification combined with intraocular lens implantation from May to December 2020 in Drum Tower Hospital, the Affiliated Hospital of Nanjing University Medical School.The 156 specimens were divided into 312 small pieces.Of the 312 pieces, 157 pieces were divided into normal control group (23 pieces), positive control group (10 pieces), balanced salt solution (BSS) immersion group (61 pieces) and distilled water immersion group (63 pieces) using computer-generated random numbers.Normal control group received no treatment.Positive control group was directly fixed with a mass fraction of 4% histiocytes fixative solution.For the 61 pieces in BSS immersion group, 20 pieces were soaked for 1 minute, 21 pieces for 2 minutes, and 20 pieces for 3 minutes.For the 63 pieces in distilled water immersion group, 20 pieces were soaked for 1 minute, 23 pieces for 2 minutes, and 20 pieces for 3 minutes.Another 125 pieces were selected to simulate the cataract aspiration-irrigation according to the treatment in BSS immersion group and distilled water immersion group respectively, plus rinse in a bottle containing BSS at a height of 70 cm for 1 minute.Cell viability was detected by trypan blue-eosin staining.LECs density, dead cell count, cell death rate and percentage of shedding (%) were calculated.Of the remaining 30 pieces, every 15 pieces were divided into normal control group, BSS immersion group, and distilled water immersion for 1, 2 and 3 minutes groups, with 3 pieces in each group.BSS immersion group was immersed for 3 minutes, and the other four groups were treated as mentioned above, and the LECs structure of the four groups was observed by light microscopy and transmission electron microscopy.This study adhered to the Declaration of Helsinki.The study protocol was approved by the Ethics Committee of Drum Tower Hospital, the Affiliated Hospital of Nanjing University Medical School (No.2019-248-01). Written informed consent was obtained from each subject.Results:The boundaries of LECs in BSS treatment groups were clear, and there was no significant difference in morphology compared with normal control group.With time increasing, LECs in distilled water treatment groups gradually swelled, and the boundaries of dead cells were not clear.There were significant differences in LECs density, dead LECs count and LECs mortality ( F=13.459, 98.918, 130.600; all at P<0.001). The LECs density was lower in 2-minute and 3-minute distilled water treatment groups than in normal control group, showing statistically significant differences (both at P<0.05). The dead LECs count and LECs mortality were higher in 1-minute, 2-minute and 3-minute distilled water treatment groups than in normal control group and BSS treatment groups for the same time, and the differences were statistically significant (all at P<0.05). Only a few shed LECs were seen in normal control group, 1-minute, 2-minute and 3-minute BSS treatment groups, and BSS immersion combined rinse group.After different time of soaking, there were more shed LECs in distilled water immersion combined rinse group, and the range of LECs shedding increased with the extension of distilled water immersion.There was a significant difference in the shedding percentage of LECs among different groups ( F=123.670, P<0.001). The shedding percentages of LECs at different time points were higher in distilled water immersion groups and distilled water immersion combined rinse groups than in normal control group, and the difference was statistically significant (all at P<0.05). The shedding percentage of LECs increased significantly in distilled water immersion groups with the extension of immersion.Light microscopy showed that the cells were destroyed in 1-minute, 2-minute and 3-minute distilled water treatment groups, and some LECs shed in the 2-minute and 3-minute treatment groups.Transmission electron microscopy showed cell lysis and destruction, suborganelles swelling, disruption of intercellular junctions in 1-minute, 2-minute and 3-minute distilled water treatment groups, loose attachment between cells and capsule in the 2-minute and 3-minute treatment groups, and cell detachment from capsule in the 3-minute treatment group. Conclusions:Distilled water immersion leads to LECs death in a time-dependent manner, and distilled water immersion combined with rinse can remove LECs on the lens capsule.

Objective:To investigate the inhibitory effect of indocyanine green (ICG) on biological behavior and transdifferentiation of human lens epithelial cells (HLECs) and its mechanism.Methods:HLECs were divided into blank control group, 5% glucose solution (GS) group and 0.5% ICG group, 1.5% ICG group and 2.5% ICG group, which were treated with balanced salt solution, 5% GS and 0.5%, 1.5% and 2.5% ICG solutions for 3 minutes, respectively, and then were incubated in fresh medium for 24 hours.The apoptosis level of HLECs was detected by flow cytometry.The expression levels of apoptosis-related proteins, Bcl-2-associated X protein (Bax), B-cell lymphoma-2 (Bcl-2), caspase-3 and caspase-9 were detected by Western blot.Cell proliferation was detected via the cell counting kit-8 (CCK-8) assay and 5-ethynyl-2'-deoxyuridine (EdU) incorporation assay.The migration ability of HLECs was detected by cell scratch assay.Cell migration and invasion were determined by Transwell assays.The expression levels transdifferentiation-related proteins, α-smooth muscle actin (α-SMA), nerve calcium adhesion protein (N-cadherin), fibronectin (FN) and vimentin were assessed by Western blot.Results:The apoptosis rates of blank control group, 5% GS group, 0.5% ICG group, 1.5% ICG group and 2.5% ICG group were (4.35±0.60)%, (4.63±0.19)%, (8.17±0.69)%, (13.90±0.33)% and (23.08±1.12)%, with a statistically significant difference in the overall comparison ( F=412.74, P<0.05). The apoptosis rate was significantly higher in 0.5% ICG group, 1.5% ICG group and 2.5% ICG group than in blank control group and 5% GS group (all at P<0.05). The relative expressions of caspase-3, caspase-9 and Bax proteins were significantly higher in 0.5% ICG group, 1.5% ICG group and 2.5% ICG group than in blank control group and 5% GS group, and the relative expression of Bcl-2 protein was lower in 1.5% ICG group and 2.5% ICG group than in blank control group and 5% GS group, and the differences were statistically significant (all at P<0.05). The rate of EdU-positive cells was significantly lower in 0.5% ICG group, 1.5% ICG group and 2.5% ICG groups than in blank control group and 5% GS group (all at P<0.05). The survival rate of cells was significantly lower in 0.5% ICG group, 1.5% ICG group and 2.5% ICG group than in blank control group and 5% GS group (all at P<0.05). The migration rates of scratch cells were significantly lower in 0.5% ICG group, 1.5% ICG group and 2.5% ICG group than in blank control group and 5% GS group, and the differences were statistically significant (all P<0.05). The number of migrating cells and the number of invading cells were significantly lower in 0.5% ICG group, 1.5% ICG group and 2.5% ICG group than in blank control group and 5% GS group (all at P<0.05). The relative expressions of α-SMA, N-cadherin and FN were significantly lower in 0.5% ICG group, 1.5% ICG group and 2.5% ICG group than in blank control group and 5% GS group, and the relative expression of vimentin was lower in 1.5% ICG group and 2.5% ICG group than in blank control group and 5% GS group, and the differences were statistically significant (all at P<0.05). Conclusions:ICG can promote HLECs apoptosis and inhibit HLECs proliferation, migration, invasion and transdifferentiation in a concentration-dependent manner.