A chiral LC-MS/MS method for the simultaneous analysis of desvenlafaxine (DVS) enantiomers in human plasma was developed and applied to a pharmacokinetic study on 12 Chinese healthy volunteers. d6-Desvenlafaxine was used as internal standard (IS). Chromatographic separation was performed on the Astec Chirobiotic V chiral column (150 mm x 4.6 mm, 5 μm). The assay was linear over the concentration range of 0.500-150 ng x mL(-1) for both enantiomers (r2 > 0.99). The method was successfully applied to a stereoselective pharmacokinetic study of 100 mg desvenlafaxine sustained release tablets on 12 Chinese healthy volunteers under fasting conditions. The results showed that the pharmacokinetic parameters were similar to both enantiomers in Chinese healthy volunteers. The AUC(0-t), and C(max) of the two enantiomers were about 1.5 times higher than those of blacks and whites reported in the literature.

Objective:To investigate the expression of miR-92a in colorectal cancer (CRC) and its effect on the regulation mechanisms of tumor angiogenesis. Methods:The miRNA-92a expression in 25 CRC tissues and HT29, SW620, SW480, and HCT116 CRC cells was detected using quantitative real-time polymerase chain reaction. The CD31 positive expression of blood microvessels in CRC tissues was measured by immunohistochemistry. Pearson correlation analysis was used to analyze the relationship between miR-92a and tu-mor angiogenesis. The miR-92a mimic or inhibitor was transfected into HCT116 and SW620 cells in vitro to upregulate or downregu-late the miR-92a expression. The effect of miR-92a overexpression or suppression on the formation of human umbilical vein endotheli-al cell (HUVEC) tubules was detected by tube formation assay. Changes in phosphatase and tensin homolog (PTEN) protein expression were measured by Western blot. Results:The expression level of miR-92a in CRC tissues was significantly higher than that in matched adjacent tissues (P<0.01). The expression levels of miR-92a in HT29, SW480, SW620, and HCT116 colon cancer cell lines were signifi-cantly higher than that of the normal colorectal epithelium control (P<0.05). The number of CD31 positive expression of microvessel density (MVD) in CRC tissues was significantly higher than that in adjacent tissues (P<0.01), and the miR-92a expression level was posi-tively correlated with the MVD in CRC tissues (r=0.580, P=0.01). The cell culture supernatant of HCT116 with miR-92a overexpression could significantly promote the formation of HUVEC tubules (P<0.05). The upregulation of miR-92a expression could significantly inhib-it the expression of PTEN protein in CRC cells (P<0.01). Conclusion:The miR-92a was highly expressed in CRC cells and tissues, which was closely related to the formation of tumor angiogenesis in CRC. The miR-92a could promote tumor angiogenesis in CRC by inhibit-ing PTEN expression.

To investigate the effects of metallothionein (MT) on isolated rat heart, 16 Wistar rats were randomly divided into 2 groups. In control group (group C), distilled water was injected intraperitoneally and 24 h later isolated hearts were perfused with Langendorff and stored at 4 degrees C for 3 h with histidine-tryptophan-ketoglutarate (HTK) solutions, and then isolated hearts were perfused for 2 h by Langendorff. In experimental group (group E), 3.6% ZnSO(4) was injected intraperitoneally, 24 h later isolated hearts were perfused by Langendorff and stored at 4 degrees C for 3 h with HTK solutions, and then the isolated hearts were perfused for 2 h with Langendorff. MT content, the recovery of hemodynamics, myocardial water content (MWC), lactate dehydrogenase (LDH) and creatine kinase (CK) leakage, adenosine triphosphate (ATP) and malondialdehyde (MDA) content, superoxide dismutase (SOD) activity, myocardial cell Ca(2+) content, Ca(2+)-ATPase activity of mitochondria ([Ca(2+)-ATPase](m)) and its Ca(2+) content ([Ca(2+)](m)), synthesizing ATP activity of mitochondria ([ATP](m)), and the ultrastructure of cells were examined. There were a significant increase in group E in hemodynamic recovery, ATP content, SOD activity, [Ca(2+)-ATPase](m) activity, [ATP](m) activity, and substantial reduction in MWC, LDH and CK leakage, MDA content, myocardial cell Ca(2+) content, [Ca(2+)](m) content, and the ultrastructural injury were obviously milder than that of group C. This study demonstrated that MT has protective effects on isolated rat heart.
Cardiotonic Agents/*pharmacology ; Creatine Kinase/*metabolism ; L-Lactate Dehydrogenase/metabolism ; Metallothionein/biosynthesis ; Metallothionein/*pharmacology ; Myocardium/*metabolism ; Myocardium/ultrastructure ; Random Allocation ; Rats, Wistar ; Superoxide Dismutase/metabolism ; Zinc Sulfate/pharmacology

To investigate the effects of metallothionein (MT) on isolated rat heart, 16 Wistar rats were randomly divided into 2 groups. In control group (group C), distilled water was injected intraperitoneally and 24 h later isolated hearts were perfused with Langendorff and stored at 4℃ for 3 h with histidine-tryptophan-ketoglutarate (HTK) solutions, and then isolated hearts were perfused for 2 h by Langendorff. In experimental group (group E), 3.6% ZnSO4 was injected intraperitoneally, 24 h later isolated hearts were perfused by Langendorff and stored at 4℃ for 3 h with HTK solutions, and then the isolated herts were perfused for 2 h with Langendorff. MT content, the recovery of hemodynamics, myocardial water content (MWC), lactate dehydrogenase (LDH) and creatine kinase (CK) leakage, adenosine triphosphate (ATP) and malondialdehyde (MDA) content, superoxide dismutase (SOD) activity, myocardial cell Ca2+ content, Ca2+-ATPase activity of mitochondria ([Ca2+-ATPase]m) and its Ca2+ content ([Ca2+]m), synthesizing ATP activity of mitochondria ([ATP]m), and the ultrastructure of cells were examined. There were a significant increase in group E in hemodynamic recovery, ATP content, SOD activity, [Ca2+-ATPase]m activity, [ATP]m activity, and substantial reduction in MWC, LDH and CK leakage, MDA content, myocardial cell Ca2+ content, [Ca2+]m content,and the ultrastructural injury were obviously milder than that of group C. This study demonstrated that MT has protective effects on isolated rat heart.

Objective:To investigate the relationship between thyroid iodine uptake rate, 99Tc m imaging and treatment of Graves' hyperthyroidism with 131I. Methods:132 patients with Graves' hyperthyroidism were analyzed retrospectively. According to the difference of thyroid 24-hour iodine uptake rate, they were divided into group A (≤50%), group B (50%-80%) and group C (≥80%). According to the trend of iodine uptake rate curve, they were divided into peak advance group (6 h/24 h iodine uptake rate ratio >1) and non-peak advance group (6 h/24 h iodine uptake rate ratio≤1). Thyroid 99Tc m imaging was divided into three groups according to quality, including group 1 (≤30 g), group 2 (30-60 g) and group 3 (≥60 g). The therapeutic effects of 131I in different types of patients were compared. Results:132 patients were followed up for 6 months after the first 131I treatment. The total effective rate, total cure rate and uncured rate were 88.6%(117/132), 78.0%(103/132), 22.0%(29/132), respectively. The serum thyroxine levels of patients with different 24-h iodine uptake rates in the three groups after treatment were significantly lower than those before treatment ( P<0.05). The lower the 24-h iodine uptake rate of thyroid, the more significantly the serum total triiodothyronine (TT3), total triiodothyronine (TT4), free triiodothyronine (FT3) and free thyroxine (FT4) levels decreased after treatment, the more obvious the increase of thyroid stimulating hormone (TSH), and the more obvious the treatment effect ( P<0.05). The cure rate (13/29, 44.8%) in the peak advance group was significantly lower than that in the non peak advance group (90/103, 87.4%), with statistically significant difference (χ 2=23.899, P<0.001). There was significant difference in the cure rate among the three groups with different thyroid 99Tc m imaging quality (χ 2=15.502, P<0.001). Conclusions:With the increase of thyroid mass, the higher the 24-h iodine intake rate, the more obvious the peak shift, the lower the cure rate, the higher the non-healing rate, and the lower the incidence of hypothyroidism.

ObjectiveTo investigate the effect of HBsAg on the expression of interferon-α (IFN-α) in peripheral blood plasmacytoid dendritic cells (pDCs) induced by the stimulator of interferon genes (STING) signaling pathway activated by cyclic GMP-AMP (cGAMP). MethodPeripheral venous blood was collected from healthy adults and the patients with chronic hepatitis B virus (HBV) infection who attended the outpatient service or were hospitalized in Department of Infectious Diseases, The Affiliated Hospital of Xuzhou Medical University, from February to December 2016, and peripheral blood mononuclear cells (PBMCs) were isolated and extracted. After the STING agonist cGAMP was added to PBMCs, ELISA was used to measure the levels of IFN-α, interferon-β, and tumor necrosis factor-α in supernatant. PBMCs from healthy adults were pre-incubated with HBsAg and then stimulated by cGAMP, and supernatant was collected to measure IFN-α. The magnetic-activated cell sorting method was used to remove pDCs from PBMCs, and after culture with cGAMP, ELISA was used to measure the level of IFN-α in supernatant. PBMCs from healthy adults were stimulated by HBsAg and/or cGAMP, and then flow cytometry was used to measure the frequency of pDCs. The independent samples t-test was used for comparison of continuous data between two groups. ResultsPBMCs from the patients with chronic HBV infection stimulated by cGAMP in vitro had a significantly lower level of IFN-α than healthy controls (469.72±18.95 vs 599.90±84.06, t=4.868, P=0.001). PBMCs from healthy adults co-cultured with HBsAg and stimulated by cGAMP had a significantly lower level of IFN-α than those in the non-HBsAg group (448.5±52.0 vs 571.0±30.8, t=4.500, P=0.011). Compared with PBMCs containing pDCs, PBMCs without pDCs stimulated by cGAMP had a significant reduction in the level of IFN-α (164.50±40.73 vs 339.50±35.33, t=6.482, P=0.001). Compared with PBMCs from healthy adults stimulated by cGAMP, PBMCs pre-incubated with HBsAg and then stimulated by cGAMP had a significant reduction in the frequency of pDCs (0.12%±0.04% vs 0.24%±0.04%, t=5.176, P=0.014). ConclusionHBsAg can inhibit the expression of IFN-α induced by the STING pathway in pDCs activated by cGAMP.

Objective:Aimed to investigate the value of adrenocorticotropic hormone (ACTH) stimulation in adrenal venous blood sampling (AVS).Methods:Patients who diagnosed as primary aldosteronism (PA) and completed successful bilateral cannulation judged by selection index (SI) for routine and(or) ACTH stimulation AVS were enrolled. The lateralization index(LI) was calculated to compare the effect of ACTH stimulation on AVS cannulation success rate and lateralization judgment.Results:A total of 73 patients with PA were enrolled in the study, of whom 28 were confirmed as aldosterone producing adenoma (APA) after unilateral adrenalectomy. Cortisol and aldosterone in peripheral and adrenal veins were significantly increased after ACTH stimulation. The left SI was increased from 6.5(3.0-13.6) to 26.8 (16.9-40.3) ( P<0.01) and the right SI from 20.8(4.8-34.8) to 57.6(35.7-80.9) ( P<0.01) after ACTH stimulation. There was no significant difference on LI before and after ACTH stimulation [7.7(2.3-19.6) vs 5.6(1.9-14.6), P=0.14]. The success rates of left and right adrenal cannulation were increased by 15% and 10% respectively after ACTH stimulation. For 57 patients who were determined in successful cannulation by both routine and ACTH stimulation AVS, 27 patients were determined to have lateralization by both AVS methods, 21 patients were determined to have bilateralization, and the consistency of lateralization by both AVS methods was 84%(48/57). Among the 28 patients who were confirmed to be APA after unilateral adrenalectomy, the correct rate of lateralization by both AVS methods was 89% (25/28). Conclusion:ACTH stimulation is able to improve the success rate of bilateral adrenal vein cannulation, and is helpful to judge AVS results. For patients with successful cannulation, there is no significant difference in lateralization judgment for routine and ACTH stimulation AVS.

Objective To evaluate the effect of microRNA-92a (miR-92a) on regulating cell proliferation by targeting Krüppel-like factor 4 (KLF4) in colon cancer.Methods The miR-92a expressions in 21 colon cancer tissues and matched normal tumor-adjacent tissues and 4 colon cancer cells (HT29,SW480,SW620,HCT116) were detected using quantitative real-time polymerase chain reaction (qRT-PCR).Models of over-expression and suppression of miR-92a were established by transient transfection of miR-92a-3p mimic to HCT116 and transient transfection of miR-92a-3p inhibitor to SW620,respectively.Cell proliferation activity was detected by the CCK-8 colorimetry method,cell cycles were detected by flow cytometry,KLF4 protein expression was detected by Western blotting,and cell luciferase activity was detected by the dual luciferase reporter gene experiment.Results The expression level of miR-92a in colon cancer tissues was (0.648 ± 0.489) fmol/μg total RNA,significantly higher than that in matched normal tumor-adjacent tissues [(0.064 ± 0.062) fmol/μg total RNA],with statistically significant difference (t =-5.420,P ＜ 0.001).In 4 colon cancer cell lines,the miR-92a expression level in HCT116 cells was the lowest,and highest in SW620 cell.When the expression of miR-92a was up-regulated,the cell proliferation activity of 72 h in HCT116 cells was higher than that in the negative control group (0.919 ±0.014 vs.0.765 ± 0.025),with statistically significant difference (t =-9.309,P =0.001),the proportion of S phase cells was also significantly increased [(41.670 ±0.461)％ vs.(38.703 ±0.554)％,t =-7.127,P =0.002),and KLF4 protein expression was decreased (0.460 ± 0.048 vs.0.758 ± 0.109,t =22.865,P =0.028).When the expression of miR-92a was down-regulated,the cell proliferation activity of 72 h in SW620 cells was lower than that in the negative control group (0.608 ± 0.011 vs.0.713 ± 0.005),with statistically significant difference (t =15.920,P ＜ 0.001),while the proportion of S phase cells was decreased [(31.935 ± 0.365) ％ vs.(34.955 ± 0.465) ％,t =8.849,P =0.001],and KLF4 protein expression was increased (0.694 ± 0.121 vs.0.479 ± 0.044,t =-5.246,P =0.034).KLF4 3'UTR wild-type dual luciferase report plasmids were co-transfected with miR-92a-3p mimic to HCT116 cell,and dual luciferase assay showed that miR-92a slightly repressed firefly luciferase actively,but the difference was not statistically significant (t =0.878,P =0.429).There was a negative correlation between the expression of miR-92a and the expression of KLF4 protein in colon cancer tissues,but with no statistical significance (r =-0.163,P =0.699).Conclusion miR-92a is highly expressed in colon cancer tissues.It can promote colon cancer cells proliferation via enhancement of the cell cycle transition of G0-G1 phase to S phase.Up-expression of miR-92a may play a role in down-regulating the expression of KLF4 protein in colon cancer cells.However,KLF4 is not a direct target gene of miR-92a.

Sleep deprivation (SD) has a profound impact on the central nervous system, resulting in an array of mood disorders, including depression and anxiety. Despite this, the dynamic alterations in neuronal activity during sleep deprivation have not been extensively investigated. While some researchers propose that sleep deprivation diminishes neuronal activity, thereby leading to depression. Others argue that short-term sleep deprivation enhances neuronal activity and dendritic spine density, potentially yielding antidepressant effects. In this study, a two-photon microscope was utilized to examine the calcium transients of anterior cingulate cortex (ACC) neurons in awake SD mice in vivo at 24-hour intervals. It was observed that SD reduced the frequency and amplitude of Ca2+ transients while increasing the proportions of inactive neurons. Following the cessation of sleep deprivation, neuronal calcium transients demonstrated a gradual recovery. Moreover, whole-cell patch-clamp recordings revealed a significant decrease in the frequency of spontaneous excitatory post-synaptic current (sEPSC) after SD. The investigation also assessed several oxidative stress parameters, finding that sleep deprivation substantially elevated the level of malondialdehyde (MDA), while simultaneously decreasing the expression of Nuclear Factor erythroid 2-Related Factor 2 (Nrf2) and activities of Superoxide dismutase (SOD) in the ACC. Importantly, the administration of gallic acid (GA) notably mitigated the decline of calcium transients in ACC neurons. GA was also shown to alleviate oxidative stress in the brain and improve cognitive impairment caused by sleep deprivation. These findings indicate that the calcium transients of ACC neurons experience a continuous decline during sleep deprivation, a process that is reversible. GA may serve as a potential candidate agent for the prevention and treatment of cognitive impairment induced by sleep deprivation.

Objective:To investigate the ultrasonographic features and evolution of thyroid shrinking nodules in order to improve the differential diagnosis and management strategy and avoid unnecessary biopsy.Methods:A total of 245 patients with old bleeding of benign thyroid nodules diagnosed via fine needle aspiration cytology (FNAC) from May 2015 to July 2020 in the southern part of the Shanghai General Hospital Affiliated to the Medical College of Shanghai Jiaotong University, including 263 nodules. The sonographic parameters such as size, shape, aspect ratio, echo, edge, boundary, periphery, halo, calcification, posterior echo enhancement and posterior acoustic attenuation of nodules were analyzed retrospectively, and the ultrasonic appearances of nodules were classified; 41 nodules from 40 patients, who did ultrasound examinations more than twice and had both complete ultrasonographic data, were compared and then classified in order to explore the evolution rules of the nodules.Results:Two hundred and sixty-three thyroid shrinking nodules in 245 cases were classified into 4 types: "cystic wall shrinkage sign" type(71.48%, 188/263), "carcinoma-like" type (22.05%, 58/263), "inflammation-like" type(3.04%, 8/263) and "undefined atypical solid nodules" type(3.42%, 9/263). Of the 41 consecutive follow-up nodules, 37 cases showed obvious cystic wall shrinkage sign, and 7 of them developed into "carcinoma-like" type and 6 cases into "inflammation-like" type.Conclusions:The ultrasonographic appearance of thyroid shrinking nodules is a dynamic process, which can be divided into 4 types: "cystic wall shrinkage sign" , "carcinoma-like" , "inflammation-like" and "undefined atypical solid nodules" types. The "cystic wall shrinkage sign" type is typical and common. The "cystic wall shrinkage sign" type can develope into the "carcinoma-like" type or the "inflammation-like" type.