Objective:To explore if there was a correlation between TNFAIP8 and gastric tumor drug resistance. Methods:We collected 47 cases of neoadjuvant chemotherapy,and detected the expression of TNFAIP8 in their tissues before and after surgery by im-munohistochemistry. Compared the expression of TNFAIP8 between SGC7901 cell and its cisplatin resistance cell SGC7901-DDP by Western blot,and then we used siRNA technology to decreased its expression,continued to test the susceptibility to cisplatin through MTT. Results:The expression of TNFAIP8 between pre-neoadjuvant and after-neoadjuvant had no difference, but the expression of chemotherapy effectiveness was lower than chemotherapy invalidness(P<0. 05);the expression of SGC7901-DDP cell was also higher than SGC7901 cell; the expression of TNFAIP8 decreased apparently through plasmid transfection of SGC7901-DDP cell, MTT test showed the fewer expression of TNFAIP8 could increase the susceptibility to cisplatin(P<0. 05). Conclusion: TNFAIP8 was involved in chemotherapy resistance of gastric carcinoma.

Objective To investigate the expression and clinicopathological significance of metastasis-associated 1 (MTA1), matrix metalloproteinase-9 (MMP-9) and vascular endothelial growth factor-C (VEGF-C) in a gastric adenocarcinoma.Methods The expressions of MTA1, MMP-9, and VEGF-C were detected with immunohistochemical method in 120 cases of gastric adenocarcinoma tissues and 30 cases of normal gastric tissues.Results The positive percentage of MTA1, MMP-9, and VEGF-C proteins in gastric adenocarcinomas was 76.7％ , 71.7％ , and 64.2％ , respectively, which was significantly higher than that (33.3％, 36.7％, and 30.0％) in normal gastric mucosa tissues (P ＜0.05).The expressions of MTA1, MMP-9, and VEGF-C were significantly higher in gastric carcinoma invasion serous membrane, lymph node metastasis, clinical Ⅲ ～ than that in non-invasion serous membrane, non-lymph node metastasis, and clinical Ⅰ ～ Ⅱ (P ＜ 0.05).The expression of MTA1 had significantly positive correlation with that of MMP-9 (P ＜ 0.05) and VEGF-C (P ＜ 0.05) in gastric adenocarcinomas.Conclusions The expressions of MTA1, MMP-9, and VEGF-C were higher in gastric adenocarcinomas than normal gastric mucosa tissues.They may play an important role in the invasion and metastasis of gastric carcinomas.MTA1 may promote infiltration, invasion, and lymph node metastasis of gastric carcinoma through regulation of the expressions of MMP-9 and VEGF-C.The combined detection of MTA1, MMP-9, and VEGF-C expressions has important value to judge the grade malignance of gastric adenocarcinomas.

Objective To investigate the effects of rhG-CSF on the improvement of cognitive impairment and anti-apoptosis of Alzheimer disease (AD) induced by Aβ1-42 in rat. Methods Healthy male Wistarirats were randomly assigned to the Aβ group, treatment group and sham operation group. Aβ1-42 (10μg) was injected into bilateral hippocampus to create the rat model of AD. Rats in rhG-CSF group were subcutaneously subjected to 50μg/(kg · d) rhG-CSF for 5 days, while rats in Aβ group were subjected to normal saline. Morris water maze tests were done and expressions of caspase-3 protein were determined by immunohistochemical method on the 7th, 14th, 21st, and 28th day after administration. Results (1) The avoiding latent periods of rhG-CSF group ( ( 34. 33 ±6. 47 ) s, (42. 08 ± 6. 36 ) s, (46. 88 ± 7. 66 ) s, respectively ) were shorter than that of Ap group ((49.79 ±4.87)s, (50.25 ±6.81 )s, (51. 33 ±6.90)s, respectively). The percentages of swimming distances in the target quadrant in rhG-CSF group ( (41.00 ±7.62)% ,(43.33 ±8. 16)% ,(44. 67 ±8.07)% ,respectively) were increased comparing with Ap group((25.33 ±6.89)% , (23. 83 ±4.67)% ,(21.50 ±4.64)% ,respectively). The differences were all statistically significant (P<0.05). (2) Compared with sham operation group,the positive rate of caspase-3 protein in rat's hippocampus of Ap group significantly increased after injecting Aβ1-42 The positive rates of caspase-3 protein in rhG-CSF group on the 7th, 14th day ( (7. 93 ±6. 33) and (8. 83 ±5. 94) were lower than Aβ group ( ( 10.43 ±7. 16) and ( 11. 34 ± 5. 17 ) . The differences were statistically significant (P< 0.05). Conclusion RhG-CSF can improve the cognitive impairment of Alzheimer Disease (AD) induced by Aβ1-42 in rat, decrease the apoptosis of hippocampal neurons and delay the decline of its learning and memory ability to some extent.

[ ABSTRACT] AIM:To observe the effect of ginsenoside Rb 1 on the proliferation and the expression of serotonin transporter (SERT), 5-hydroxytryptamine 1B receptor (5-HT1BR) in rat pulmonary artery smooth muscle cells (PASMCs) under hypoxia condition and the relationship with Rho /Rho-kinase signal pathway .METHODS: PASMCs were isolated from the adult male SD rats and primarily cultured .The subcultured cells from the 4th generation to the 6th generation were harvested and divided into normal group , and hypoxia group , different concentrations of Rb 1 incubation groups treated with 50, 100 and 200 mg/L ginsenoside Rb1 under hypoxia (HR50, HR100 and HR200 groups, respectively).The viability of the PASMCs was measured by CCK-8 assay.BrdU positive cells were determined using flow cytometry .The expression of serotonin transporter and 5-HT1BR at mRNA and protein levels was detected by RT-PCR and Western blot, respectively. The PASMCs were randomly divided into normal group , hypoxia group , HR200 group and hypoxia +Y-27632 incubation group ( HY group ) .The mRNA expression of Rho-kinase and phosphorylated myosin phosphatase target subunit 1 ( p-MYPT1) protein level were investigated by RT-PCR and Western blot, respectively.RESULTS: Compared with normal group, the proliferation of PASMCs in hypoxia group was significantly increased (P<0.01).The cell viability and the ex-pression of SERT and 5-HT1B R at mRNA and protein levels in all different concentrations of Rb 1 groups were obviously de-creased compared with hypoxia group ( P<0.05 ) .The mRNA expression of Rho-kinase and protein level of p-MYPT1 were markedly decreased in HR200 group, and no significant difference compared with HY group was observed ( P <0.01).CONCLUSION: Treatment with ginsenoside Rb1 might prevent hypoxia-induced proliferation of PASMCs and over-expression of SERT and 5-HT1B R through inhibiting the Rho/Rho-kinase pathway .

Objective To determine the feasibility of magnetically labeling and tracking mesenchymal stem cells(MSCs)in vitro by using a gadolinium and fluorescent bi-functionally transfection agent of polyethylenimine.Methods A gadolinium bifunctional transfection reagent complex was obtained after the linear polyethylenimine derivative(JetPEI-FluoR)was incubated with Gd-DTPA.Mesenchymal stem cells isolated from the bone marrows of SD rats were cultured and expanded.The mesenchymal stem cells were incubated with the bi-functional labeling agents.After labeling,the MSCs were examined with fluoroscope and electron microscope and the biological characters were detected including trypan blue exclusion test,MTT,and apoptosis detection.On a 1.5 T MR system,the labeled MSCs were examined with spin echo T1 WI and T2 WI and T1 measurement with mixed sequence.After labeling,the cells were cultured and undergone routine passage.Prior MR examinations were repeated for each passage of labeled cells.All data was statistically prolessed with SPSS for Windows.Results Of 5×105 MSCs incubated with the bi-functional agents,4.25×105 MSCs were successfully labeled,the percentage of labeled MSCs was 85% fluoroscopically.The high density electron particles of gadolinium observed electron microscopically existed around cellular apparatuses,especially around Golgi apparatus.In trypan blue exclusion test,the exclusion rate of labeled MSCs with incubation duration of 3,6,12,24 h was(96.55±2.90)%,(94.17±2.56)%,(97.16±3.12)% and(94.23±2.67)%,respectively.The corresponding exclusion rate of unlabeled MSCs was(95.86±2.67)%,(92.04±2.21)%,(93.38±3.64)%and(92.12±2.53)%,respectively.There was no statistical difference of trypan blue exclusion rate between labeled cells and control unlabeled cells within 24 hours of incubation(F=4.523,P＞0.05).In the proliferation test,the optical absorption value of labeled MSC with 2.5,5.0,10.0,20.0,30.0 and 40.0 μl bi-functional labeling agent was(0.1884±0.0151),(0.1878±0.0190),(0.1741±0.0160),(0.1135±0.0215),(0.1079±0.0145)and(0.0811±0.0079),respectively.The corresponding optical absorption value of unlabeled MSCs was(0.1940±0.0116).The optical absorption value of labeled cells was not affected in case of less than 30.0 μl of Gd-DTPA(q'=0.2225-0.9458,P＞0.05).The apoptosis index for labeled cells and unlabeled cells were 5.08% and 3.86%,respectively.On T1 WI,the signal intensity and T1 relaxation time of unlabeled cells and labeled cells were 240.3±24.7 and(2457±56)ms,336.2±20.7 and(1102±64)ms,respectively,and there were significant statistical difference(t=12.656,17.889,P＜0.01).The minimal amount of cells which was detectable for T1 WI was 5×103.After routine passage,the gadolinium in the cells gradually decreased and could be tracked by MRI until the fifth passage.Conclusions The gadolinium and fluorescent bi-functionally labeling rat bone marrow mesenchymal stem cell by using the transfection agent of polyethylenimine is feasible,efficient and safe.The labeled cells could be tracked in vitro on MR imaging.

OBJECTIVETo assess the association of neural precursor cell expressed developmentally down-regulated 4 (NEDD4) with schizophrenia.

METHODSFive single nucleotide polymorphisms (SNPs) of the NEDD4 gene were genotyped by TaqMan SNP genotyping assay in an independent sample of 464 individuals with schizophrenia and 487 healthy controls from eastern Han Chinese population. Clinical data were collected with a general information questionnaire and Positive and Negative Syndrome Scale (PANSS).

RESULTSFrequencies of rs3088077 (allelic: χ2=18.024, P=0.000; genotypic: χ2=16.634, P=0.000), rs7162435 (allelic: χ2=6.771, P=0.009; genotypic: χ2=7.352, P=0.025) and rs2303579 (allelic: χ2=11.253, P=0.001; genotypic: χ2=12.248, P=0.002) were found to be significant different between the two groups. Moreover, TT of rs7162435 was significantly correlated with scores of factors of excitement and hostility (14.53±3.925, F=3.551, P=0.029).

CONCLUSIONrs3088077, rs7162435 and rs2303579 of the NEDD4 gene may be associated with schizophrenia. Moreover, the TT genotype of rs7162435 may increase the severity of excitement and hostility. Our results may provide a clue for delineating the connection between the glutamate hypothesis of schizophrenia and ubiquitination.

Adolescent ; Adult ; Aged ; Alleles ; Asian Continental Ancestry Group ; ethnology ; genetics ; China ; ethnology ; Endosomal Sorting Complexes Required for Transport ; genetics ; Female ; Genetic Predisposition to Disease ; Genotype ; Humans ; Male ; Middle Aged ; Nedd4 Ubiquitin Protein Ligases ; Polymorphism, Single Nucleotide ; Schizophrenia ; enzymology ; ethnology ; genetics ; Ubiquitin-Protein Ligases ; genetics ; Young Adult

Objective:To explore the clinical value of heparin binding protein (HBP) in bronchoalveolar lavage fluid (BALF) for diagnosis and differential diagnosis of bacterial pneumonia.Methods:Eighty eight patients with pulmonary infection from the respiratory department of the Fourth Affiliated Hospital of Anhui Medical University from January 2019 to January 2021 were enrolled in the study, including 48 cases of bacterial pneumonia and 40 cases of non-bacterial pneumonia; meanwhile, 40 non-pulmonary infection patients were also enrolled as the control group. The BALF levels of HBP, procalcitonin (PCT), interleukin-6 (IL-6) were measured, and the clinical values of the above indexes in differential diagnosis of bacterial and non-bacterial pneumonia were analyzed.Results:The BALF levels of HBP and IL-6 in bacterial pneumonia group were significantly higher than those of the non-bacterial pneumonia group and the control group ( P<0.05). ROC curve showed that the areas under the curve (AUC) of HBP and IL-6 were 0.930 and 0.893 for the early diagnosis of bacterial pneumonia; and the sensitivity was 88.5% and 82.7%, the specificity was 92.5% and 92.5%, respectively. Combined detection of HBP and IL-6, the AUC was 0.942 and the sensitivity was 94.2% and the specificity was 95.0%. When they were used to distinguish bacterial pneumonia, the AUC of HBP and IL-6 were 0.890 and 0.777, and the sensitivities were 80.8% and 71.2%, and the specificity were 91.7% and 75.0%, respectively. Combined detection of HBP and IL-6, the AUC was 0.902, and the sensitivity was 96.2% and the specificity was 79.2%. Conclusions:BALF HBP and IL-6 have good clinical value in the early diagnosis and distinguishing bacterial pulmonary infection and the joint value of the two is better.

Objective:To investigate the iodine nutritional status of pregnant women in Qingdao and the effect of prevention and treatment of iodine deficiency disorders (IDD), so as to provide a basis for residents to supplement iodine scientifically, and take targeted prevention measures and adjust intervention strategies.Methods:In accordance with the requirements of the "National Iodine Deficiency Disorders Surveillance Program (2016 edition)" and "Shandong Iodine Deficiency Disorders Surveillance Program", the cluster sampling method was adopted to select pregnant women from 10 districts (cities) in Qingdao from 2018 to 2020, to investigate their basic information and thyroid disease history. Meanwhile, household edible salt samples and random urine samples were collected to detect iodine content.Results:A total of 3 000 pregnant women were monitored from 2018 to 2020, the median age was 31 years, and the median gestational age was 18 weeks. There were significant differences in the distribution of age, gestational age, whether senile puerpera, and pregnancy in different years ( H/χ 2 = 29.35, 81.03, 65.62, 77.34, P < 0.001). The median salt iodine of edible salt ( n = 3 000) and iodized salt ( n = 2 700) in pregnant women's homes were 23.02 and 23.70 mg/kg, respectively. The qualified rate of iodized salt, the coverage rate of iodized salt and the consumption rate of qualified iodized salt were 89.59% (2 419/2 700), 90.00% (2 700/3 000) and 80.63% (2 419/3 000). The comparison of qualified rate of iodized salt, coverage rate of iodized salt and consumption rate of qualified iodized salt among different years was statistically significant (χ 2 = 48.09, 36.62, 61.08, P < 0.001), the coverage rate of iodized salt and the consumption rate of qualified iodized salt showed a downward trend year by year (χ 2trent = 35.54, 29.50, P < 0.001). A total of 3 000 urine samples were collected from pregnant women and the median urinary iodine of pregnant women was 147.85 μg/L. The urinary iodine level in the third trimester was lower than that in the first and second trimesters ( P < 0.001). The urinary iodine level in the non elderly group was higher than that in the elderly group ( Z = - 6.66, P < 0.001). The urinary iodine level in the group without thyroid disease was higher than that in the group with thyroid disease ( Z = - 1.99, P = 0.047). The urinary iodine level in iodized salt group was higher than that in non-iodized salt group ( Z = - 2.42, P = 0.015). Conclusions:The iodine nutrition of pregnant women in Qingdao is generally at an insufficient level, and the risk of iodine deficiency is high, which needs attention. In recent years, the coverage rate of iodized salt and the consumption rate of qualified iodized salt in Qingdao have shown a downward trend, and have failed to meet the requirements of national standards. In the future, we should strengthen the monitoring and health education of IDD in pregnant women.

Objective:To explore the mechanism of highly expressed primary cilia in tibial growth plate chondrocytes accelerating chondrocytes differentiation in young rats with chronic renal failure (CRF).Methods:Forty male 4-week-old SD rats weighing (98±3) g were randomly divided into control group (intragastric administration with distilled water, n=20) and CRF group (intragastric administration with adenine suspension 150 mg·kg -1·d -1, n=20). All the young rats were sacrificed after continuous gavage for 6 weeks. The length of the growth plate was measured with histological sections. Immunofluorescence (IF) was used to detect the expression rate of primary cilia and the level of β-catenin, the key protein of Wnt/β-catenin signaling pathway in tibial growth plate chondrocytes. Chondrocytes isolated from growth plate in two groups were cultured in vitro to P3 generation, and the expression rate of primary cilia in chondrocytes, the levels of Indian hedgehog (IHH) and glycogen synthase kinase 3β (GSK3β) were detected by IF. Co-immunoprecipitation was used to detect the relationship between IHH and GSK3β. Results:Compared with the control group, the relative length of the growth plate was shorter in histological sections [(0.51±0.11) vs (1.00±0.08), t=16.11, P<0.001], the expression rate of primary cilia was higher [(26.3±5.5)% vs (7.6±1.9)%, t=14.37, P<0.001], and the level of β-catenin increased [(7.1±2.0) scores vs (3.6±1.0) scores, t=7.10, P<0.001] in CRF group. In vitro, the expression rate of primary cilia was higher in CRF group chondrocytes [(31.4±8.2)% vs (12.5±3.1)%, t=9.64, P<0.001] than that in control group. The level of IHH in CRF group increased than that in control group [(1 360±270) vs (310±84), t=16.61, P<0.001]. There was no significant difference in GSK3β level of chondrocytes between the two groups [(850±195) vs (780±140), t=1.30, P=0.200]. There was a direct interaction between IHH and GSK3β in CRF group chondrocytes. Conclusions:The expression levels of primary cilia and related protein IHH increase in tibial growth plate chondrocytes of CRF young rats. The IHH protein plays a direct interaction with GSK3β protein, Wnt/β-catenin signaling pathway antagonist, which leads to the activation of Wnt/β-catenin signaling pathway and final accelerated differentiation of chondrocytes. The rapid differentiation of chondrocytes causes the closing trend of growth plate.

Objective To establish new classification criteria for early rheumatoid arthritis (E-RA) based on large samples of early inflammatory arthritis patients and to evaluate the value of this criteria in China.Methods Patients who had arthritic complaints with disease duration less than one year were enrolled.They were divided into RA group and non-RA group according to the clinical diagnosis by experienced rheumatologists.The clinical and laboratory parameters were analyzed and those with high sensitivity or specificity were selected as the new classification criteria.Statistical analysis was carried out by using t test,x2 test and Logistic regression.Results ① A total of 803 patients with early inflammatory arthritis were included in this study.Five hundreds and fourteen patients were diagnosed as early RA and 251 were diagnosed as other rheumatic diseases,and the diagnosis of 38 patients remained unestablished by the end of follow-up.② New E-RA classification criteria were established based on the parameters with high sensitivity and/or specificity.The sensitivity of the new E-RA criteria was 84.4％,which was higher than 1987 ACR criteria (58.0％),while the corresponding specificities were similar,which were 87.4％ and 93.6％ respectively.③ Compared with the complex scoring system of 2010 ACR/EULAR criteria,the E-RA criteria was more simple and practical.The diagnostic sensitivity and specificity of E-RA criteria were higher than those of 2010 ACR/EULAR criteria reported in the literatures.④ New classification criteria based on scoring system using Logistic regression analysis was established.The sensitivity of this criteria was 86.4％,which was higher than 1987 ACR criteria (58.0％).Conclusion The diagnostic value of the E-RA criteria developed in this study for early RA is better than 1987 ACR criteria,and is more simple than 2010 ACR/EULAR criteria.It may be used as a new classification criteria for early RA diagnosis.