Objective To observe the variation of phosphatidylinositol-3 kinase (PI3K),protein kinase B (AKT),sodium iodide symporter (NIS) mRNA and protein expression in rat mammary tissues and serum insulin growth factor Ⅰ (IGF-1) under different iodine nutrition levels,and to study the role of PI3K-AKT signaling pathway in the process of mammary gland intaking iodine during lactation period.Methods Totally 130 Wistar rats (100 female rats,30 male rats) were randomly divided into five groups with 20 female rats in each group:①control group (NI):was feed with normal diet and iodine content 50 μg/L in deionized water;②low iodine group 1 (LI1 group):was feed with low iodine diet and deionized water;③low iodine group 2 (LI2):was feed with low iodine diet and iodine content 5 μg/L in deionized water;④high iodine group 1 (HI1 group):was feed with normal diet and iodine content 3 000 μg/L in deionized water;⑤high iodine group 2 (HI2):was feed with normal diet and iodine content 10 000 μg/L in deionized water.After feeding for 3 months,females were mated with male rats,then male rats were taken out and every female rat was feed individually.Urinary iodine level of rats in lactation period 10 days after giving birth was tested.Blood and mammary tissue samples of rats in lactation period were taken after killing them.Enzyme linked immunosorbent assay (ELISA) was used to detect serum IGF-1 level,real-time fluorescence quantification PCR to detect the mRNA expression of mammary gland PI3K,AKT and NIS,Western blotting to detect mammary gland PI3K,total AKT,phosphorylation AKT (p-AKT) and NIS protein expression.Results The medians urinary iodine of lactation period rats in LI1 and LI2 (3.16,6.36 μg/L) were significantly lower than that in NI group (162.59 μg/L),and were significantly higher in HI1 and HI2 (2 356.27,11 507.29 μg/L) than that in NI group.The differences were statistically significant (all P ＜ 0.01).Compared with control group [(8.84 ± 2.12) μg/L],the content of serum IGF-1 increased significantly in lactation period rats in LI1 and LI2 groups [(13.30 ± 2.37) and (10.90 ± 1.92) μg/L,all P＜ 0.01].The real-time fluorescence quantification PCR detection results indicated that the differences were statistically significant by comparing NIS,AKT,PI3K mRNA expression of the mammary tissues of lactation period rats in the five groups (F=14.916,36.477,14.994,all P＜ 0.01).Among them,NIS mRNA expression quantities in LI1 and LI2 groups (0.75 ± 0.40,0.89 ± 0.51) were significantly higher than that in NI group (0.53 ± 0.31),and significantly lower in HI2 group (0.30 ± 0.24) than that in NI group (P ＜ 0.05 or ＜ 0.01).AKT mRNA expression quantities in LI1 and LI2 groups (0.90 ± 0.19,0.64 ± 0.22) were significantly higher than that in NI group (0.43 ± 0.22),and significantly lower in HI2 group (0.29 ± 0.15) than that in NI group (P ＜ 0.05 or ＜ 0.01).PI3K mRNA expression quantity in LI1 group (0.85 ± 0.42) was significantly higher than that in NI group (0.50 ± 0.24),and significantly lower in HI2 group (0.28 ± 0.10) than that in NI group (all P ＜ 0.01).Western blot detection results indicated that the differences were statistically significant by comparing mammary gland NIS protein expression of lactation period rats in the five groups (F=4.510,P＜ 0.01).Among them,LI1 group (1.67 ± 0.97) was significantly higher than NI group (0.87 ± 0.43,P ＜ 0.05).The differences were statistically significant by comparing the p-AKT protein expression among groups (F =3.528,P ＜ 0.05).Among them,HI2 group (1.10 ± 0.30) was significantly higher than NI group (0.75 ± 0.23,P ＜0.05).The differences were not statistically significant by comparing total AKT and PI3K protein expression among groups (F =0.558,1.319,all P ＞ 0.05).Conclusion The inhibitory effect of PI3K-AKT signaling pathways on NIS in the mammary gland was weaker than the effect of iodine intake.But the expression of functional p-AKT was gradually increased with the increment of iodine intake,which had been presented inhibit effect on NIS expression in lactating mammary gland.

Objective:To investigate the influence of T2DM Spleen-qi Deficiency syndrome on Ankle-Brachial Index (ABI).Methods:The clinical data of 298 patients with T2DM who were hospitalized in Dongzhimen Hospital, Beijing University of Chinese Medicine from January 2019 to December 2021 were retrospectively analyzed. According to the diagnostic criteria of spleen-qi deficiency syndrome, the patients were divided into two groups: spleen-qi deficiency syndrome group and non-spleen-qi deficiency syndrome group. There were 142 patients in the spleen-qi deficiency syndrome group and 156 patients in the non-spleen-qi deficiency syndrome group. The differences of ABI between the two groups were compared, and the correlation between spleen-qi deficiency syndrome and clinical indicators (gender, age, body mass index, course of diabetes, history of hypertension, smoking history, fasting glucose, total cholesterol, triglyceride, platelet, hemoglobin, serum albumin, blood urea nitrogen, serum creatinine, estimated glomerular filtration rate) and ABI in T2DM patients was analyzed.Results:The left ABI [1.09 (1.00, 1.19) vs. 1.13 (1.03, 1.22)] and the right ABI [1.09 (0.96, 1.17) vs. 1.12 (1.02, 1.20)] in T2DM spleen-qi deficiency syndrome group were significantly lower than those in non-spleen-qi deficiency group ( P<0.05).The left ABI was significantly correlated with spleen-qi deficiency syndrome ( r=0.122, P=0.035) and estimated glomerular filtration rate ( r=0.137, P=0.018), and the right ABI was significantly correlated with spleen-qi deficiency syndrome ( r=0.123, P=0.034) and PLT ( r=-0.115, P=0.047). After correcting for other confounding factors by multiple linear regression analysis, there was significantly correlation between spleen-qi deficiency syndrome and ABI. Conclusion:Compared with the non-spleen-qi deficiency syndrome group, T2DM patients in the spleen-qi deficiency group had a lower ankle-brachial index and were more likely to develop peripheral arterial disease.

Objective To investigate the Th1/Th2 cytokines and chemokine CXCL 13 levels in peripheral blood and cerebrospinal fluid(CSF)of patients with neurosyphilis and their clinical significance. Methods Forty seven HIV negative patients with neurosyphilis, 36 syphilis patients without neurological involvement(syphilis group)and 23 patients without infectious intracranial diseases(control group) admitted in Hangzhou Third Hospital during July 2011 to December 2014 were enrolled in the study.CSF pressure, protein contents, white blood cell counts and IgG index were detected in patients with neurosyphilis.Enzyme-linked immunosorbent assay(ELISA)was used to detect the levels of Th1 cytokines (IL-2,IL-12 and IFN-γ),Th2 cytokines(IL-6,IL-10)and CXCL13 in serum and CSF in three groups.Th1/Th2 cytokines,CXCL13 levels and CSF routine were also examined in neurosyphilis patients after treatment for 3 and 12 months.ANOVA, Kruskal-Wallis test and Spearman correlation were used for data analysis.Results The levels of IL-6 and IL-10 in serum and CSF from neurosyphilis patients were higher than those in syphilis group and control group(χ2IL-6=15.43, 15.39 and 14.44, 20.01,χ2 IL-10 =16.46, 23.86 and 15.11,24.44;P<0.05 or <0.01), while the levels of IL-2, IL-12 and IFN-γin the serum and CSF were lower than those in syphilis group and control group(χ2IL-2=14.55,17.14 and 16.14,17.97;χ2IL-12=13.65,20.50 and 18.48,21.04;χ2IFN-γ=16.95,17.53 and 16.00,15.21;P<0.05 or <0.01). CXCL13 contents in CSF of neurosyphilis patients were significantly higher than those in other two groups (χ2=52.51 and 53.76, P <0.01), and were positively correlated with leucocyte counts, protein concentrations,IgG index,IL-6 and IL-10(r=0.325,0.544,0.750,0.333 and 0.382,P<0.05 or <0.01),but were negatively correlated with IL-12 levels in neurosyphilis patients(r=-0.303,P<0.05). In neurosyphilis patients,CXCL13 and IL-6 levels were reduced after treatment for 12 months(χ2=102.00 and 22.17,P<0.05 and <0.01), while the levels of IL-2 and IL-12 were increased(χ2=18.28 and 24.10,P<0.05 and <0.01).Conclusion Chemokine CXCL13 and Th1/Th2 cytokines are involved in the immune response in neurosyphilis patients, which may be used for the diagnosis and evaluation of therapeutic efficacy in patients with neurosyphilis.

OBJECTIVE: To investigate the persistent damage of learning and memory ability after the cessation of sub-chronic manganese(Mn)-exposure in rats. METHODS: Specific pathogen free weaning male SD rats were randomly divided into control group and low-, medium-and high-dose groups based on body weight, with 6 rats in each group. Rats were intraperitoneally injected with Mn chloride(MnCl_2·4 H_2O) at the concentrations of 0, 5, 10, or 20 mg/kg body weight, 5 days per week for 6 weeks and continued to be observed for 12 weeks after the cessation of Mn-exposure. During the experiment, the body mass of the rats was weighed. Learning and memory ability was evaluated by a Morris water-maze task at the 6 th weeks of Mn-exposure(cessation of Mn-exposure of week 0), the 6 th and 12 th week of the cessation of Mn-exposure. The organ coefficients of heart, liver, spleen, kidney and testicles were evaluated after the cessation of Mn-exposure on week 12. RESULTS: The body mass of the high-dose group was lower than that of the other 3 groups(P<0.05) at the 4 th and 6 th week of Mn-exposure and the 2 nd week of the cessation of Mn-exposure. There was no significant difference in body mass between the groups(P>0.05) on the 12 th week of the cessation of Mn-exposure. The escape latency of high-dose group was higher than that of the control group(P<0.05), and the number of platform crossings in the low-, medium-and high-dose groups were fewer than that in the control group(P<0.05) after the cessation of Mn-exposure. The escape latency was shorter and the numbers of platform crossings were higher on the 6 th and 12 th week of the cessation of Mn-exposure(P<0.05) when compared with that of the 6 th week of Mn-exposure rats. There was no statistical significance in the organ coefficients of heart, liver, spleen, kidney and testicles among the 4 groups at the 12 th week of the cessation of Mn-exposure in rats(P>0.05).CONCLUSION: Sub-chronic Mn exposure can impair learning and memory ability of rats, and the damage persists after the cessation of Mn-exposure.

Objective To develop an ultra-high performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) method to simultaneously detect the contents of gallic acid, syringin, phellodendrine, aesculin and rhein in the gallnut lotion. Methods An UPLC- MS/MS method was established. Separation was performed on an Agilent Poroshell 120 EC-C₁₈(2.1 mm×150 mm, 2.7 μm)with a gradient mobile phase system of 0.2% formic water-acetonitrile solution. The flow rate was 0.3 ml/min. The temperature of column was 30 ℃. The injection volume was 2 μl. The MS detection was in dynamic MRM mode. Results gallic acid, syringin, phellodendrine, aesculin and rhein were successfully separated using this method, with good linear relationship as the ranges of 153.8-15380、10.31-1031、5.265-526.5、50.70-5070、1.054-105.4 ng/ml, respectively. The precision, repeatability, stability and recovery were good. Conclusion This UPLC-MS/MS method is stable, rapid, and reproducible., It is suitable for detecting the contents of gallic acid, syringin, phellodendrine, esculetin and in the gallnut lotion.

OBJECTIVE: To investigate the effect of osthole on the expression of amyloid precursor protein (APP) in Alzheimer's disease (AD) cell model and its mechanism. METHODS: The SH-SY5Y cell with over expression of APP was established by transfection by liposome 2000. The cells were treated with different concentrations of osthole, and the cell viability was determined by MTT and lactate dehydrogenase (LDH) assay. The differentially expressed miRNAs with and without osthole treatment were detected by miRNA array, and the target genes binding to the differentially expressed miRNAs were identified and verified by databases and Cytoscape. After the inhibitor of the differentially expressed miRNA was transduced into cells, the changes of APP and amyloid β (Aβ) protein were determined by immunofluorescence cytochemistry, and the mRNA expression of APP was determined by RT-PCR. RESULTS: The AD cell model with over expression of APP was established successfully. The results of MTT and LDH assay showed that osthole had a protective effect on cells and alleviated cell damage. miR-101a-3p was identified as the differentially expressed miRNA, which was binding to the 3'-UTR of APP. Compared with APP group, the expression of APP and Aβ protein and APP mRNA increased in the miR-101a-3p inhibitor group (all <0.01), while the expression of APP and Aβ protein and APP mRNA decreased in the cells with osthole treatment (all <0.01). CONCLUSIONS Osthole inhibits the expression of APP by up-regulating miR-101a-3p in AD cell model.
Alzheimer Disease ; Amyloid beta-Peptides ; Amyloid beta-Protein Precursor ; genetics ; Cell Line ; Coumarins ; pharmacology ; Gene Expression Regulation ; drug effects ; genetics ; Humans ; MicroRNAs ; genetics ; metabolism

Objective: To observe the influence of lower wide pedicle frontal periosteum flap on frontal bone absorption rate after cranioplasty. Methods: From February 2016 to July 2017, the lower wide pedicle frontal periosteum flap was produced in 12 patients of Shanghai Ninth People′s Hospital (7 males/5 females, aged 5-9 years, 10 hypertelorism, 2 Crouzon syndrome), who accepted intro-cranio-route plastic surgery, to cover the frontal bone window. A spiral CT scans were applied one week (t1) and one year (t2) after surgery. DICOM data was imported into Mimics software to reconstruct the 3D model of skull. The bone window covered the frontal bone was selected and the bone volume was calculated. The absorption rate was calculated as (Vt₁-Vt₂)/Vt₁×100%. As a control group, the CT data of 20 patients (from January 2010 to December 2015, 11 males/9 females, 7 hypertelorism, 12 Crouzon syndrome, 1 Pfeiffer syndrome) were analyzed retrospectively in the same way, and compared to the experimental group. Results: The average bone absorption rate in experimental group was 8.65%±2.56% (n=12), while in control group it was 26.51%±5.23% (n=20). Significant statistical difference was observed. No further cranial defect was observed in one-year follow-up in both two groups. Conclusions The lower wide pedicle frontal periosteum flap reduces the bone absorption rate after intro-cranio-route plastic surgery. It could also help to repair the anterior cranial base defect during the surgery. This technique was recommend as a regular step in craniofacial surgeries.