AIM:To investigate the effects of down-regulation of protein kinase C (PKC) on the activity of storeoperated Ca2 + channels (SOC) and the proliferation of airway smooth muscle cells (ASMCs). METHODS:Rat bronchial smooth muscle cells were isolated and cultured. Fluo-3 /AM fluorescence was measured by laser confocal microscope to assessing intracellular Ca2 +. Downregulation of PKC activity was achieved by incubation of ASMCs with PKC activator phorbol-12-myristate-13-acetate (PMA,10 ?mol/L) or phorbol 12,13 -dibutyrate (PDBu,1 ?mol/L) for 24 h. The proliferation of ASMCs was assayed by calculating the reduction rates of Alamar blue. RESULTS:Down-regulation of PKC activity by longterm exposure of PMA or PDBu inhibited the proliferation of ASMCs,the similar results were obtained by using PKC inhibitor chelerythrine. Both downregulation of PKC activity and inhibition of PKC activity by chelerythrine reduced Ca2 + entry through SOC channels. Low concentration of PMA (0. 1 ?mol/L) promoted the proliferation of ASMCs,and this effect was inhibited by SOC blocker SKF-96365. CONCLUSION:Inhibition or down -regu-lation of PKC activity results in the inhibition of SOC channels,suggesting that PKC is involved in the activation of these channels. Ca2 + entry through SOC channels might contribute to PKC-promoted proliferation of ASMCs.

Store-operated Ca(2+) channels (SOCs) are plasma membrane Ca(2+) permeable channels activated by depletion of intracellular Ca(2+) store. Ca(2+) entry through SOCs is known as store-operated Ca(2+) entry (SOCE), which plays an important role in the functional regulation of airway smooth muscle cells (ASMCs). Protein kinase C (PKC) has been shown to have an activating or inhibiting effect on SOCE, depending on cell types and PKC isoforms that are involved. In ASMCs, the effect of PKC on SOCE has not been elucidated so far. In this study, the role of PKC in the activation of SOCE in rat ASMCs was examined by using Ca(2+) fluorescence imaging technique. The results showed that acute application of PKC activators PMA and PDBu did not affect SOCE induced by the sarcoplasmic reticulum Ca(2+)-ATPase (SERCA) inhibitor thapsigargin. The non-selective PKC inhibitor chelerythrine significantly inhibited thapsigargin- and bradykinin-induced SOCE. RT-PCR assay identified PKCα, δ and ɛ isoforms in rat ASMCs. PKCα-selective inhibitor Gö6976 and PKCɛ-inhibiting peptide Epsilon-V1-2 had no effect on SOCE; by contrast, PKCδ-selective inhibitor rottlerin attenuated SOCE dramatically, suggesting that PKCδ was the major PKC isoform involved in the activation of SOCE in ASMCs. Moreover, PKC down-regulation by extended exposure to high doses of PMA or PDBu also reduced SOCE, confirming the essential role of PKC in the activation of SOCE in ASMCs. In addition, PKC down-regulation did not influence the expression of stromal interaction molecule 1 (STIM1) and Orai1, two elementary molecules in the regulation and activation of SOCs. These results identified PKCδ as an essential PKC isoform involved in the activation of SOCE, and confirmed that PKC regulates the function of ASMCs in a SOCE-dependent manner.

Store-operated Ca(2+) channels (SOCs) are plasma membrane Ca(2+) permeable channels activated by depletion of intracellular Ca(2+) store. Ca(2+) entry through SOCs is known as store-operated Ca(2+) entry (SOCE), which plays an important role in the functional regulation of airway smooth muscle cells (ASMCs). Protein kinase C (PKC) has been shown to have an activating or inhibiting effect on SOCE, depending on cell types and PKC isoforms that are involved. In ASMCs, the effect of PKC on SOCE has not been elucidated so far. In this study, the role of PKC in the activation of SOCE in rat ASMCs was examined by using Ca(2+) fluorescence imaging technique. The results showed that acute application of PKC activators PMA and PDBu did not affect SOCE induced by the sarcoplasmic reticulum Ca(2+)-ATPase (SERCA) inhibitor thapsigargin. The non-selective PKC inhibitor chelerythrine significantly inhibited thapsigargin- and bradykinin-induced SOCE. RT-PCR assay identified PKCα, δ and ɛ isoforms in rat ASMCs. PKCα-selective inhibitor Gö6976 and PKCɛ-inhibiting peptide Epsilon-V1-2 had no effect on SOCE; by contrast, PKCδ-selective inhibitor rottlerin attenuated SOCE dramatically, suggesting that PKCδ was the major PKC isoform involved in the activation of SOCE in ASMCs. Moreover, PKC down-regulation by extended exposure to high doses of PMA or PDBu also reduced SOCE, confirming the essential role of PKC in the activation of SOCE in ASMCs. In addition, PKC down-regulation did not influence the expression of stromal interaction molecule 1 (STIM1) and Orai1, two elementary molecules in the regulation and activation of SOCs. These results identified PKCδ as an essential PKC isoform involved in the activation of SOCE, and confirmed that PKC regulates the function of ASMCs in a SOCE-dependent manner.
Animals ; Bronchi ; metabolism ; Calcium ; metabolism ; Calcium Channels ; Calcium Signaling ; physiology ; Cells, Cultured ; Male ; Membrane Glycoproteins ; metabolism ; Myocytes, Smooth Muscle ; metabolism ; ORAI1 Protein ; Protein Kinase C-delta ; metabolism ; Rats ; Rats, Sprague-Dawley ; Stromal Interaction Molecule 1

Objective:To observe the changes of retinal blood flow density and thickness in the macular region of eyes with high myopia (HM) combined with peripapillary intrachoroidal cavitation (PICC).Methods:A cross-sectional study. From March 2019 to May 2021, 65 patients (65 eyes) diagnosed as PICC (HM+PICC group) in Eye Hospital, China Academy of Chinese Medical Sciences, sex-and age-matched 69 HM patients of 69 eyes (HM group) and 65 healthy people of 65 eyes (control group) were enrolled in this study. The optical coherence tomography angiography was used to scan macular areas in 3 mm×3 mm, and measure the macular fovea and optic disc on superior, inferior, nasal, temporal superficial capillary plexus (SCP) and deep capillary plexus (DCP) vessel density in the foveal and parafoveal region, and macular retinal ganglion cell complex (mGCC) thickness, full retinal thickness. One-way analysis of variance were used to test the difference of the index values among three groups, and then two groups were compared with Bonferroni test. A paired t-test was used to test the difference of the macular vessel density and thickness between the superior and inferior hemifield in three groups. Pearson partial regression analysis was used to calculate the correlations between them at same sites. Results:PICC was located most frequently at the inferior temporal disc border, followed by the inferior nasal region, superior temporal region, and superior nasal region in the HM+PICC group on 57(87.7%, 57/65), 25(38.5%, 25/65), 3(4.6%, 3/65) and 1(1.5%, 1/65 ) eye. There were significant differences in the global and regional full retinal thickness, mGCC thickness, SCP and DCP vessel density among 3 groups ( F=29.097, 51.929, 16.253, 6.135; P<0.001). The macular SCP and DCP vessel density except in the fovea, all regional macular full retinal thickness and mGCC thickness in the HM+PICC group were significantly lower than those in the normal group ( P<0.05). Compared to the HM group, the HM+PICC group had lower all regional mGCC thickness and SCP vessel density, as well as full retinal thickness in the inferior hemifield and DCP vessel density in the foveal region ( P<0.05). Macular vessel density and thickness in the inferior hemifield were significantly lower than those in the superior hemifield ( t=6.356, 11.693, 6.212, 2.936; P<0.01). Pearson partial regression analysis showed the SCP vessel density was positively correlated with corresponding mGCC thickness and full retinal thickness ( r=0.584, 0.534, 0.592, 0.496, 0.485, 0.517; P<0.001). However, there was no significant correlation between the DCP vascular density and mGCC thickness ( P>0.05), and only a weak positive correlation between the DCP vascular density and the full retinal thickness in the inferior hemifield ( r=0.319, P=0.014). However, no association with average and superior full retinal thickness ( r=0.066, 0.002, 0.125, 0.184, 0.016, 0.319; P>0.05). Conclusion:The macular SCP vessel density, mGCC thickness and the full retinal thickness in the inferior hemifield in PICC eyes are lower than those in the HM eyes, especially the mGCC thickness and SCP vessel density in the inferior hemifield, and there is a strong positive correlation between them.

OBJECTIVEThe aim of this study was to detect the plasma concentration of OLC1 (overexpressed in lung cancer 1) protein as a potential cancer biomarker, and evaluating its clinical application value in the diagnosis of non-small cell lung cancer (NSCLC).

METHODSWe prepared OLC1 antibody with OLC1 full length protein, in 5-6-week old Bal B/c mice. Each mouse was immunized four times at a dose of 15-30 µg antigen protein, and the interval between two consecutive immunizations was two weeks. Antibody screening was made by ELISA and Western blot, and a double antibody sandwich ELISA kit was developed. We used this established ELISA kit to detect the plasma concentration of OLC1 protein in 281 NSCLC patients and 92 gender- and age-matched healthy controls. Area under the receiver operating characteristic curve (AUC) was used to evaluate the detection efficacy of OLC1.

RESULTSWe obtained 11 OLC1 monoclonal antibodies and successfully established the ELISA kit to detect the plasma concentration of OLC1 with a detection range from 1.95 ng/ml to 62.50 ng/ml. OLC1 concentration in the case group (124.69 ng/ml) was significantly higher than that in the control group (67.07 ng/ml, P < 0.001). In the scenario of distinguishing NSCLC from control group, AUC result was 0.69. When the cut-off was set at 67.72 ng/ml, the sensitivity and specificity was 84.4% and 51.1%, respectively. In term of distinguishing early lung cancer (IA) from normal controls, the AUC, sensitivity and specificity were 0.68, 77.8% and 54.4%, respectively.

CONCLUSIONThe plasma concentration of OLC1 protein is significantly elevated in NSCLC patients. OLC1 may be as a potential cancer biomarker applied in clinical diagnosis.

Adult ; Animals ; Antibodies, Monoclonal ; Biomarkers, Tumor ; blood ; Blotting, Western ; Carcinoma, Non-Small-Cell Lung ; blood ; diagnosis ; immunology ; Early Detection of Cancer ; methods ; Enzyme-Linked Immunosorbent Assay ; Female ; Humans ; Lung Neoplasms ; blood ; diagnosis ; immunology ; Male ; Mice, Inbred BALB C ; Middle Aged ; Oncogene Proteins ; blood ; immunology ; ROC Curve ; Sensitivity and Specificity ; Young Adult

Objective:To analyze the mental health status of men who have sex with men (MSM) population among acquired immunodeficiency syndrome (AIDS) patients.Methods:A face-to-face questionnaire survey was conducted to investigate the general information, symptom check list-90 (SCL-90) questionnaire and scale of 269 MSM patients with AIDS in the Eighth Hospital of Xi'an city from November 2015 to May 2016.Results:Among the 269 cases of MSM with human immunodeficiency virus (HIV)/AIDS, the majority were 20-40 years old, with higher education level, mostly unmarried and nonsingleton. The scores of each factor of SCL-90 were significantly higher than that of the norm, among which diet and sleep, interpersonal relationship, anxiety and depression were the most prominent; regression analysis showed that the main reason affecting the scores of each factor among AIDS patients was the low quality of life, and the patients with high education were more likely to have depression and anxiety.Conclusions:MSM has its own special characteristics among AIDS patients, and there are many psychological problems. Necessary psychological intervention measures should be given.

Objective:To investigate the local consistency of inferior colliculus and ventrolateral orbital cortex by resting-state functional magnetic resonance imaging (fMRI) in rats with noise induced deafness and its relationship with anxiety- and depression-like behavior.Methods:Twenty-four clean grade male four-weeks old SD rats were randomly divided into noise group and control group with 12 rats in each group.Rats in the noise group were exposed to 122 dB broadband strong noise for 2 hours to induce severe bilateral hearing loss, while rats in the control group were placed in a quiet environment. Hearing thresholds were assessed by auditory brainstem response (ABR) test. The open field test (OFT) was conducted to examine anxiety-depression related behavior, and the local consistency in the rat brain was evaluated by fMRI.SPM12 software was used to process fMRI data, and Pearson correlation analysis was conducted by SPSS 22.0 software to calculate the correlation between fMRI data and behavior.Results:The results of ABR showed that the full band hearing threshold of rats in the noise group was higher than that of rats in the control group ((85.417±6.463) dB, (20.083±8.853) dB, t=46.168, P<0.001). And compared with control group, the rats in the noise group showed obvious anxiety-depression-like behavior in the open field test, that was, low activity level.The results of OFT showed that the total distance ((39.912±5.696) m, (47.993±10.820)m, t=-2.289, P=0.032), average moving speed ((13.306±1.900)cm/s, (15.998±3.607)cm/s, t=-2.290, P=0.032) and standing times ((13.333±5.960), (23.500±7.323), t=-3.730, P=0.001) of the rats in the noise group were all lower than those in the control group. Compared with the control group, the local consistency of hypothalamus in the noise group was significantly enhanced, while the local consistency of ventrolateral orbital cortex was significantly reduced, and the abnormal neural activity was lateralized. The correlation analysis showed that the neural activity of the inferior colliculus was negatively correlated with the total distance of rats in the noise group moving in the open field( r=-0.691, P=0.013), while the neural activity of the ventrolateral orbital cortex was not significantly correlated with the anxiety-depression-like behavior in the open field. Conclusions:The neural activity of inferior colliculus is closely related to anxious-depression behavior in rats with noise-induced deafness, while the ventrolateral orbital cortex may be related with other behaviors.

OBJECTIVE：To establish the fingerprint of Adiantum capillusveneris from different producing areas ，and to conduct chemometric analysis and content determination of differential components ，so as to provide reference for quality control of A. capillusveneris . METHODS ：HPLC-DAD combined with Similarity Evaluation System of TCM Chromatogramtic Fingerprint （2004 A edition ）were used to establish fingerprint of 19 batches of A. capillusveneris from different producing areas （S1-S19）. Common peaks were confirmed and their similarities were evaluated. Chemometric analysis methods such as cluster analysis ， principle component analysis （PCA），orthogonal partial least squares discriminant analysis （OPLS-DA）were used to evaluate the quality of 19 batches of A. capillusveneris from different producing areas ，screen the differential components ，and determine the contents of some differential components. RESULTS ：Among 19 batches of A. capillusveneris from different producing areas ，22 common peaks were confirmed ；peak 9 was chlorogenic acid ，peak 17 was quercetin- 3-O-β-D-glucopyranoside，peak 20 was kaempferol-3-O-rutoside；the similarity of 19 batches of sample were 0.677-0.962. Through cluster analysis ，it was found that S 7 and S 10 were clustered into one category ；S15 and S 18 were clustered into one category ；and S 1-S6，S8，S9，S11-S14，S16，S17 and S 19 were clustered into one category. PCA and OPLS-DA found that S 7 and S 10 were clustered into one category ；S15 were clustered into one category ；S18 were clustered into one category ；S1-S6，S8，S9，S11-S14，S16，S17 and S 19 were clustered into one category. Chlorogenic acid ，quercetin-3-O-β-D-glucopyranoside，kaempferol-3-O-rutoside and chemical composition represented by peak 14 were the differential components of the〔2017〕2841）； medicinal material. Among 19 batches of A. capillusveneris ， average contents of chlorogenic acid and quercetin- 3-O-β-D- glucopyranoside and kaempferol- 3-O-rutoside were 0.10-4.25， 0.31-7.11，0.61-12.00 mg/g，respectively. CONCLUSIONS ： 电话：0851-86614212。 HPLC-DAD fingerprints of A. capillusveneris from different producing areas are establishe d in the study ，and three common peaks are identified. Four differential components affecting the quality of A. capillusveneris are screened ， and the contents of chlorogenic acid ， quercetin-3-O-β-D-glucopyranoside and kaempferol-3-O-rutoside in A. capillusveneris from different producing areas were significantly different.