Discovery and verification of anti-inflammatory-related quality markers in the aerial part of Bupleurum scorzonerifolium by UPLC-Q-TOF-MS/MS and in RAW 264.7 cells and a zebrafish model.

INTRODUCTION: The root of Bupleurum scorzonerifolium Willd. (BS) is officially recognized in the Chinese Pharmacopoeia. In contrast, the aerial part of BS (ABS), accounting for 80% of BS, is typically discarded, causing potential waste of medicinal resources. ABS has shown benefits in the treatment of inflammation-related diseases in China and Spain, and the material basis underlying its anti-inflammatory effects must be systematically elucidated for the rational use of ABS. OBJECTIVE: We aimed to screen and validate the anti-inflammatory quality markers (Q-markers) of ABS and to confirm the ideal time for ABS harvesting. METHODS: The chemical components and anti-inflammatory effects of ABS from 10 extracted parts were analyzed by UPLC-Q-TOF-MS/MS and in a lipopolysaccharide (LPS)-induced cell model. Anti-inflammatory substances were screened by Pearson bivariate analysis and gray correlation analysis, and the anti-inflammatory effects were verified in a zebrafish tail-cutting inflammation model. HPLC was applied to measure the Q-marker contents of ABS in different harvesting periods. RESULTS: Ten ABS extracts effectively alleviated the increase in LPS-induced proinflammatory cytokines in RAW 264.7 cells. Forty components were identified from them, among which 27 were common components. Eight components were correlated with anti-inflammatory effects, which were confirmed to reverse the expression of proinflammatory and anti-inflammatory factors in a zebrafish model. Chlorogenic acid, hypericin, rutin, quercetin, and isorhamnetin can be detected by HPLC, and the maximum contents of these five Q-markers were obtained in the sample harvested in August. CONCLUSION: The anti-inflammatory Q-markers of ABS were elucidated by chromatographic-pharmacodynamic-stoichiometric analysis, which served as a crucial basis for ABS quality control.

SAMD9 Promotes Postoperative Recurrence of Esophageal Squamous Cell Carcinoma by Stimulating MYH9-Mediated GSK3ß/ß-Catenin Signaling.

Recurrence is a challenge to survival after the initial treatment of esophageal squamous cell carcinoma (ESCC). But, its mechanism remains elusive and there are currently no biomarkers to predict postoperative recurrence. Here, the possibility of sterile alpha motif domain-containing protein 9 (SAMD9) as a predictor of postoperative recurrence of ESCC is evaluated and the molecular mechanisms by which SAMD9 promotes ESCC recurrence are elucidated. The authors found that the high level of SAMD9 is correlated with postoperative recurrence and poor prognosis of ESCC. Overexpression of SAMD9 promotes tumor stemness, angiogenesis, and EMT, while downregulation of SAMD9 reduced these phenotypes. Mechanistically, it is found that SAMD9 stimulated ubiquitination-mediated glycogen synthase kinase-3 beta (GSK-3ß) degradation by interaction with myosin-9 (MYH9) and TNF receptor-associated factor 6 (TRAF6), which in turn activated Wnt/ß-catenin pathway. Further, the authors demonstrated that silencing SAMD9 inhibited lung metastasis and tumor formation in vivo. Finally, the authors found that silencing MYH9 or ß-catenin, or overexpressing GSK-3ß inhibited SAMD9-stimulated ESCC cell stemness, EMT, angiogenesis, metastasis, and tumorigenicity. Together, the findings indicate that the SAMD9/MYH9/GSK3ß/ß-catenin axis promotes ESCC postoperative recurrence and that SAMD9 is a crucial target for ESCC therapy. Additionally, SAMD9 has the potential as a predictor of postoperative recurrence in ESCC.

Analysis of Microcirculation Changes in the Macular Area and Para-Optic Disk Region After Implantable Collamer Lens Implantation in Patients With High Myopia.

Myopia has become an important public health problem to be solved urgently. Posterior chamber phakic implantable Collamer lens (ICL) implantation is one of the latest and safest products for myopia correction worldwide. This prospective cross-sectional case series aimed to observe changes in the macular retinal thickness, retinal nerve fiber layer (RNFL) thickness of para-optic disk region, and blood flow density after posterior ICL implantation in patients with high myopia using optical coherence tomography angiography (OCTA). A total of 67 eyes of 67 patients with high myopia, who underwent ICL implantation at The Affiliated Eye Hospital of Nanjing Medical University from January 2020 and December 2020, were included. The spherical equivalent (SE) of the operative eyes was >-6.00 D. The changes in vision, intraocular pressure (IOP), SE, and vault were observed pre-operatively, and follow-up were performed 1 week, 1 month, and 3 months. OCTA was used to observe the changes in the CRT, retinal thickness of paracentral fovea, FAZ, superficial and deep retinal blood flow density in the macular area, RNFL thickness of para-optic disk region, and blood flow density before and after ICL implantation. The uncorrected distance visual acuity (UDVA) and best corrected distance visual acuity (CDVA) of the patients post-operation were significantly improved (P < 0.001). The IOP increased in comparison with other time points at 1 week post-operation (P < 0.05). There were no significant changes in CRT post-operation. The retinal thickness in the upper, lower, nasal, and temporal quadrants of the paracentral fovea increased significantly at 1 month and 3 months post-operation (P < 0.05). The FAZ area at all postoperative time points were decreased (P < 0.001). At 3 months post-operation, the blood flow density of the superficial and deep retinal layers in the upper, lower, and nasal macular area were significantly reduced (P < 0.05). At 1 month post-operation, the RNFL thickness in the temporal para-optic disk region and blood flow density were significantly reduced (P = 0.001 and P < 0.05, respectively). ICL implantation for highly myopic eyes led to an increase of the retinal thickness in the upper, lower, nasal, and temporal regions of the paracentral fovea; reduction of RNFL thickness in the temporal area of para-optic disk; decrease in FAZ area; and decrease in the blood flow density of some deep and superficial retinal layers as well as that of the temporal para-optic disk region.

Effect of Central Corneal Thickness on Corneal Higher Order Aberrations After Cataract Surgery.

PURPOSE: To investigate the changes of corneal higher order aberrations (HOAs) before and after phacoemulsification in patients with different central corneal thickness (CCT). METHODS: In this retrospective non-randomized, non- comparative cases series, 51 eyes of 51 patients who underwent phacoemulsification were enrolled. The corneal total HOAs and Zernike coefficients (3rd and 4th order) over the 3- and 4-mm zones were measured by the iTrace aberrometer (Tracey Technologies). Participants were divided into three groups (528 µm or less, 529 to 550 µm, and greater than 550 µm) depending on the CCT tested by the LenStar LS900 (Haag Streit AG). The corneal aberrations between groups were compared with analysis of variance, the variation of corneal aberrations before and after phacoemulsification was analyzed by the paired-samples t test, and generalized linear models were used to compare postoperative aberrations between groups. RESULTS: Significant differences were found in oblique trefoil at 3 mm and vertical trefoil, oblique trefoil, and HOAs at 4 mm in the 528 µm or less group; vertical trefoil, oblique trefoil, oblique quadrafoil, and oblique secondary astigmatism at 3 mm and oblique trefoil and oblique secondary astigmatism at 4 mm in the 529 to 550 µm group; and vertical quadrafoil at 3 mm in the greater than 550 µm group. After corrected deviation of preoperative aberrations, there were significant differences in postoperative vertical coma and oblique secondary astigmatism at 4 mm when compared to the 528 µm or less and greater than 550 µm groups. CONCLUSIONS: The lower the CCT, the greater the change in corneal aberrations in patients with cataract after phacoemulsification. Surgeons should be aware of the potential for worse visual quality related to surgically induced aberrations in patients with thin corneas. [J Refract Surg. 2021;37(12):842-847.].

The Human Positive Cofactor 4 is a Promising Chemotherapeutic Target in Lung Adenocarcinoma.

Reduced sensitivity to chemotherapeutic drugs is almost inevitable in lung adenocarcinoma patients. Thus, understanding the relevant mechanisms is urgent. Positive cofactor 4 (PC4) was at first revealed to be a coactivator of basal transcription. Previous research has shown that PC4 participates in various cellular processes in normal and malignant cells. However, it is still unknown whether PC4 participates in altering the lung adenocarcinoma cell sensitivity to chemotherapy, and the relevant mechanisms remain to be explained. In this study, we discovered that PC4 was overexpressed in cisplatin-resistant lung adenocarcinoma cells. PC4 decreased cisplatin's cytotoxic effects on lung adenocarcinoma in vivo and in vitro. Furthermore, PC4 positively correlated with SOX9 in multiple cancers. PC4 was an upstream regulator of SOX9 in lung adenocarcinoma. Furthermore, PC4 mediated lung adenocarcinoma cell sensitivity to the HIF-PH inhibitor DMOG and the mTOR inhibitor rapamycin, and PC4 mediated the synergistic effect of DMOG and cisplatin. Finally, PC4 destabilized HIF-1α upon cisplatin treatment. Our research showed that PC4 participates in mediating lung adenocarcinoma cell sensitivity to multiple drugs. Mechanistically, PC4 governs multiple downstream pathways associated with chemotherapy resistance, including the SOX9 and HIF-1α pathways. Thus, PC4 is a promising chemotherapeutic target in lung adenocarcinoma.

BCAR1 plays critical roles in the formation and immunoevasion of invasive circulating tumor cells in lung adenocarcinoma.

Background: We investigated the roles of breast cancer anti-estrogen resistance 1 (BCAR1/p130Cas) in the formation and immunoevasion of invasive circulating tumor cells (CTCs) in lung adenocarcinoma (LUAD). Methods: Biomarkers of CTCs including BCAR1 and CD274, were evaluated by the CanPatrol method. Proteomics analysis of LUAD cells and exosomes after BCAR1 overexpression (BCAR1-OE) was performed by mass spectrometry. Cell functions and relevant signaling pathways were investigated after BCAR1 knockdown (BCAR1-KO) or BCAR1-OE in LUAD cells. Lastly, in vitro and in vivo experiments were performed to confirm the roles of BCAR1 in the formation and immunoevasion of CTCs. Results: High expression of BCAR1 by CTCs correlated with CD274 expression and epithelial-to-mesenchymal transition (EMT). RAC1, together with BCAR1, was found to play an important role in the carcinogenesis of LUAD. RAC1 functioned with BCAR1 to induce EMT and to enhance cell proliferation, colony formation, cell invasion and migration, and anoikis resistance in LUAD cells. BCAR1 up-regulated CD274 expression probably by shuttling the short isoform of BRD4 (BRD4-S) into the nucleus. CTCs, as well as tumor formation, were prohibited in nude mice xenografted with BCAR1-KO cells. The co-expression of BCAR1/RAC1 and BCAR1/CD274 was confirmed in LUAD. BCAR1 expression in LUAD is an indicator of poor prognosis, and it associates with immunoevasion. Conclusion: BCAR1, as a new target for the treatment of LUAD, plays roles in the formation and immunoevasion of invasive CTCs. The mechanism includes triggering EMT via RAC1 signaling and up-regulating CD274 expression by shuttling BRD4-S into the nucleus.

FOXO1 and FOXO3a sensitize non-small-cell lung cancer cells to cisplatin-induced apoptosis independent of Bim.

Low sensitivity to chemotherapy has been a major challenge in the treatment of non-small-cell lung cancer (NSCLC). It is of great clinical significance to discover its mechanisms to improve cell sensitivity to chemotherapeutic drugs. The forkhead box subfamily O (FOXO) transcriptional factors are downstream factors of the phosphatidylinositol 3-kinase/protein kinase B (PI3K/AKT) pathway and are reported to play pro-apoptotic roles in a variety of cells including NSCLC cells. But their roles and mechanisms in mediating cell response to chemotherapy remain to be discovered. We proposed that FOXO1 and FOXO3a may increase the sensitivity of NSCLC cells to cisplatin. Moreover, we presumed that LY294002, an inhibitor of the PI3K/AKT pathway, may enhance the cytotoxic effects of cisplatin through upregulating FOXO1 and FOXO3a. In the present study, we found that cisplatin initially increased the expressions and nuclear accumulation of FOXO1 and FOXO3a in NSCLC. Knockdown of FOXO1 and FOXO3a significantly decreased the cell sensitivity to cisplatin in vitro and in vivo. Moreover, inhibition of FOXO1 and FOXO3a attenuated cisplatin-induced cell apoptosis independent of Bim, a pro-apoptotic protein downstream of the FOXOs. Moreover, LY294002 synergistically increased the cytotoxic effects of cisplatin. Mechanistically, LY294002 increased the expressions and nuclear accumulation of FOXO1 and FOXO3a. Knockdown of FOXO1 and FOXO3a abrogated the enhancing effect of LY294002 on cisplatin. Taken together, our results suggested that FOXO1 and FOXO3a sensitize NSCLC cells to cisplatin and mediate the enhancing effects of LY294002 on cisplatin.

BCAR1 promotes proliferation and cell growth in lung adenocarcinoma via upregulation of POLR2A.

BACKGROUND: This study was designed to investigate the effects of a novel carcinogenetic molecule, p130cas (breast cancer antiestrogen resistance protein 1 or BCAR1) on proliferation and cell growth in lung adenocarcinoma. The study also aimed to identify the possible underlying signal networks of BCAR1. METHODS: First, we evaluated proliferation, cell colony formation, apoptosis, and cell cycle after BCAR1 was knocked out (KO) using CRISPR-Cas9 technology in H1975 and H1299 human lung adenocarcinoma cells. Subsequently, BCAR1 was upregulated in 293T cells and immunoprecipitation-mass spectrometry (IP-MS) was used with bioinformatics analysis to screen for potential networks of BCAR1 interacting proteins. Ultimately, we validated the correlated expressions of BCAR1 and a selected hub gene, RNA polymerase II subunit A (POLR2A), in 54 lung adenocarcinoma tissues, as well as in H1975 and H1299 cells. RESULTS: Cell proliferation of H1975 and H1299 was significantly inhibited following BCAR1-KO. Colony formation of H1975 cells was also significantly decreased following BCAR1-KO. IP-MS demonstrated 419 potential proteins that may interact with BCAR1. Among them, 68 genes were significantly positively correlated to BCAR1 expression, as verified by TCGA. Six hub genes were revealed by PPI String. High expression of POLR2A, MAPK3, MOV10, and XAB2 predicted poor prognosis in lung adenocarcinoma, as verified by the K-M plotter database. POLR2A and MAPK3 are involved in both catalytic activity and transferase activity. POLR2A and BCAR1 were significantly increased in lung cancer tissues as compared with matched normal tissues. High expression of POLR2A was significantly positively correlated to BCAR1 overexpression and predicted poor prognosis in 54 lung cancer cases. POLR2A expression was significantly decreased following BCAR1-KO in H1975 and H1299 cells. CONCLUSIONS: BCAR1 promotes proliferation and cell growth, probably via upregulation of POLR2A and subsequent enhancement of catalytic and transferase activities. However, additional robust studies are required to elucidate the mechanisms involved.

Evaluation of SARS-CoV-2 neutralizing antibodies using a vesicular stomatitis virus possessing SARS-CoV-2 spike protein

SARS-CoV-2 is a novel coronavirus that emerged in 2019 and is now classified in the genus Coronavirus with closely related SARS-CoV. SARS-CoV-2 is highly pathogenic in humans and is classified as a biosafety level (BSL)-3 pathogen, which makes manipulating it relatively difficult due to its infectious nature. To circumvent the need for BSL-3 laboratories, an alternative assay was developed that avoids live virus and instead uses a recombinant VSV expressing luciferase and possesses the full length or truncated spike proteins of SARS-CoV-2. Furthermore, to measure SARS-CoV-2 neutralizing antibodies under BSL2 conditions, a chemiluminescence reduction neutralization test (CRNT) for SARS-CoV-2 was developed. The neutralization values of the serum samples collected from hospitalized patients with COVID-19 or SARS-CoV-2 PCR-negative donors against the pseudotyped virus infection evaluated by the CRNT were compared with antibody titers determined from an immunofluorescence assay (IFA). The CRNT, which used whole blood collected from hospitalized patients with COVID-19, was also examined. As a result, the inhibition of pseudotyped virus infection was specifically observed in both serum and whole blood and was also correlated with the results of the IFA. In conclusion, the CRNT for COVID-19 is a convenient assay system that can be performed in a BSL-2 laboratory with high specificity and sensitivity for evaluating the occurrence of neutralizing antibodies against SARS-CoV-2.

Changes in angle kappa and angle alpha before and after cataract surgery.

PURPOSE: To observe the distributions and changes in angle kappa and angle alpha preoperatively and postoperatively in patients with cataract who were implanted with a multifocal intraocular lens (mIOL). SETTING: The First Affiliated Hospital of Northwest University, Shaanxi, China. DESIGN: Prospective nonrandomized noncomparative case series. METHODS: Eyes that underwent phacoemulsification were included. The magnitudes and orientations of angle kappa and angle alpha preoperatively and postoperatively were compared, respectively. RESULTS: The study comprised 81 eyes of 70 patients. The magnitude of angle kappa significantly decreased after phacoemulsification. No significant differences were observed between preoperative and postoperative angle kappa in orientation, as well as between preoperative and postoperative angle alpha. The correlations between preoperative and postoperative angle kappa and angle alpha were significant. There were displacement vectors for angle kappa (0.15 ± 0.10) and angle alpha (0.12 ± 0.12) after phacoemulsification. Locations of angle kappa of right and left eyes were scattered mostly in the temporal side of corneal light reflection, whereas locations of angle alpha were well organized in the horizontal position on temporal sides of corneal light reflection. CONCLUSIONS: The distribution of angle alpha was more organized compared with angle kappa. Angle kappa may change after phacoemulsification. During preoperative evaluation for patients with cataract planning for mIOLs implanted angle alpha may be a more reliable and stable factor compared with angle kappa.

The optimization and assessment of the method for inducing hyperuricemia in rats / 中国应用生理学杂志

To explore an effective method for inducing a rat model with hyperuricemia in a short period and assess the effects of the model. Methods: Sprague-Dawley rats were adopted as donors and randomly divided into control group (CT group, n=6) and 5 model groups (M1-M5 groups, n=8 in each group). M1 group (gavage with 10 g/kg yeast extracts and 100 mg/kg adenine, twice per day, 300 mg/kg oxonic acid potassium by intraperitoneal injection, in the 7 day of model inducing), M2 group (gavage with 10 g/kg yeast extracts and 100 mg/kg adenine, twice per day, 300 mg/kg oxonic acid potassium by intraperitoneal injection, in the 1, 3 and 7 day of model inducing),M3 group (gavage with 10 g/kg yeast extracts and 100 mg/kg adenine, twice per day, 300 mg/kg oxonic acid potassium by intraperitoneal injection, once per day during the model inducing), M4 group (gavage with 20 g/kg yeast extracts and 100 mg/kg adenine, twice per day, 300 mg/kg oxonic acid potassium by intraperitoneal injection, once per day during the model inducing), M5 group (gavage with 30 g/kg yeast extracts and 100 mg/kg adenine, twice per day, 300 mg/kg oxonic acid potassium by intraperitoneal injection, once per day during the model inducing), and group CT (gavaged with equal volume sterilized water and intraperitoneal injected with normal saline according to the weight and at the same frequency as the model groups). The model inducing lasted for 7 days. After the inducing was finished, blood and 24-hour urine were sampled for uric acid and creatinine determination. Then rats were maintained for 2 weeks and blood and 24-hour urine samples were collected, the concentration of uric acid and creatinine were detected. The kidney and stomach were weighed,morphological changes in kidney were observed. After model inducing, the body weight of rats in all model groups was lower than that of the control group (P＜0.01). Deaths occurred in all the rats with model treatments except M2. M4 and M5 groups were failed to be analyzed because of the high mortality, model 1 and 3 groups had 4 and 2 deaths, respectively. The uric acid levels in blood and urine of the model groups were significantly elevated (P＜ 0.01) at the end of model inducing. The model 2 group's blood uric acid was highest among the model groups (P＜0.05). It sustained a higher concentration than CT group in the three model groups after 2 weeks feeding (P＜0.05). The kidneys in model groups obviously swelling and were heavier than CT group (P＜0.01). The inflammation and structural damages were observed in kidneys of all model groups. The yeast extract (10 g/kg), adenine (100 mg/kg) gavage combined with intraperitoneal injections(the 1, 3, 7 day during inducing) of potassium oxonate can be an rapid and effective method for inducing the rat model with hyperuricemia, which can be suggested to the related research.

Analysis of medication regularity of traditional Chinese medicine prescriptions for gastropyretic excessiveness diabetes based on data mining / 中国中药杂志

To analyze the medication regularity of traditional Chinese medicine(TCM) prescriptions for gastropyretic excessiveness diabetes recorded in Chinese Medicine Prescriptions Dictionary. A total of 103 eligible prescriptions were input into the system platform, and the Apriori algorithm was used to analyze their medication regularity. The 103 prescriptions for gastropyretic excessiveness diabetes were selected from the system, and 29 herb medicines were found with frequency of usage more than 8. Totally 33 commonly used herbal pairs(support degree≥10), twenty-three 3-herb core combinations(support degree≥8, confidence values≥0.5), and twenty-one 4-herb core combinations(confidence values≥0.5) were discovered after the medication regularity analysis by Apriori algorithm. The herbal medicine combinations with the highest correlation degree were discovered after the association rule analysis on the 103 prescriptions(support degree≥10, confidence values≥0.5). The four properties, five tastes, channel distributions and frequency of dose of the 103 prescriptions were also obtained after the corresponding analysis. According to the analysis and summary of the above data, the combination of Trichosanthis Radix, Anemarrhenae Rhizoma, Coptidis Rhizoma and Ophiopogonis Radix could reflect the medication regularity of TCM prescriptions for gastropyretic excessiveness diabetes to a certain degree, which is of great significance in guiding value in clinic.

Diagnostic value of 12 leads ECG combined with detection of myocardial enzyme spectrum for atypical myocardial infarction / 中国医学装备

Objective: To explore the diagnostic value of 12 leads electrocardiogram (ECG) combined with detection of myocardial enzyme spectrum for atypical myocardial infarction. Methods: 80 patients with atypical myocardial infarction were enrolled in the observation group, and 80 patients with typical myocardial infarction in the same period were enrolled in the control group. Both of two groups were implemented the routine 12 leads ECG examination using FX-8322 ECG instrument, and the changes of Q wave and ST segment were recorded. And the series of indicators of serum myocardial enzymes (CK, CK-MB and cTnI) were detected by using AU2700 automatic biochemical analyzer. The test results of the first symptoms, ECG characteristics, myocardial enzyme spectrum and diagnostic accuracy of the two groups were compared. Results: In 80 patients of observation group, the painless first symptom was 20%, and the atypical pain was 75%, the typical upper abdominal and back pain accounted for 5%. In 80 patients of control group, all of them were typical upper abdominal and back pain, and the differences of these indicators between the two groups were significant (x2=8.889, x2=48.000, x2=72.381, P<0.05). For characteristics of ECG, the 22.5% of 80 patients of observation group hasn't been changed at ST-T segment, and the 17.5% of them was no pathological Q wave +T wave erect +ST segment elevation, and the percentage of pathological Q wave +T wave erect +ST segment elevation in observation group was 60.0%. On the other hand, the 100.0% of 80 patients of control group was pathological Q wave+T wave inverted +ST segment elevation, the differences of these indicators between the two groups were significant (x2=10.141, x2=7.671, x2=20.000, P<0.05). The results of myocardial enzyme spectrum detection indicated that the levels of serum CK, CK-MB and cTnI in the observation group were significantly lower than those in the control group (t=59.766, t=20.735, t=13.352, P<0.05), respectively. The diagnostic accuracy rate of ECG in observation group was 87.5%, which was significantly lower than 100% in control group (x2=5.333, P<0.05). The accuracy rate of ECG combined with myocardial enzyme spectrum in the observation group was 100%, which was not statistically significant compared with that in control group. Conclusion: 12 leads ECG is effective in diagnosing typical myocardial infarction, but it is necessary to combine myocardial enzyme spectrum detection for atypical myocardial infarction so as to increase the diagnostic accuracy rate.

Prokaryotic expression and preparation of the polyclonal antibodies of E protein and domain Ⅲ in Zika virus / 中国人兽共患病学报

To clone,express and purify the E Protein and EDⅢ of Zika virus in E.coli and prepare two kinds of polyclonal antibodies,the virus was amplified by Vero E6 cell culture.Total RNA was extracted by RT-PCR and reverse transcribed into cDNA.The prokaryotic expression vectors pET32a/E and pET28a/EDⅢ were constructed by cDNA sequence of E and EDⅢ gene.Then,recombinant plasmids were transformed into E.coli BL21 and induced by IPTG,and purified by Ni+ column affinity chromatography.BALB/C mice were immunized with purified recombinant proteins.Antiserum was collected and titer was determined by indirect ELISA.Western blot was used to detect the specificity.Results showed that the recombinant proteins were successfully expressed and purified.The titer of the polyclonal antibodies both reached 1:409 600.Western Blot analysis showed that the polyclonal antibodies could specifically recognize the recombinant proteins.Thus,the specific polyclonal antibody were successfully prepared,laying a foundation for further study on the pathogenesis,detection methods and immune strategies of Zika virus.

Infiltrating mast cells promote neuroendocrine differentiation and increase docetaxel resistance of prostate cancer cells by up-regulating p21 / 南方医科大学学报

<p><b>OBJECTIVE</b>To investigate the effect of infiltrating mast cells on neuroendocrine differentiation (NED) and docetaxel sensitivity of prostate cancer (PCa) cells in vitro.</p><p><b>METHODS</b>Human PCa cell lines (LNCaP and C4-2) were co-cultured with human mast cell line (HMC-1) in Transwell chambers. Androgen receptor (AR) was silenced in C4-2 cells using sh-AR lentivirus, and p21 was knocked down and overexpressed by transfecting C4-2 cells with pLKO.1-sh-p21 and pCMV-p21, respectively. The morphological changes of LNCaP and C4-2 cells were observed. MTT assay and colony formation assay were used to assess the proliferation of LNCaP and C4-2 cells. CCK8 assay was used to detect the cell viability of C4-2 cells following docetaxel trreatment. RT-qPCR and Western blotting were performed to determine the mRNA and protein expressions of neuroendocrine markers, AR and p21 in the cells.</p><p><b>RESULTS</b>Co-culture with HMC-1 cells enhanced the neuroendocrine phenotypes, inhibited the proliferation and up-regulated the expression of p21 in LNCaP and C4-2 cells. P21 positively regulated NED through a non-AR-dependent signaling pathway, while p21 knockdown partially reversed NED promoted by the mast cells. PCa cells co-cultured with HMC-1 cells showed increased resistance to docetaxel, and silencing p21 partially reversed docetaxel resistance in PCa cells.</p><p><b>CONCLUSION</b>Infiltrating mast cells up-regulates p21 to promote NED and increase docetaxel resistance in PCa cells in vitro.</p>

Case control study on Mobi-C cervical artificial disc replacement and anterior cervical decompression MC+fusion for the treatment of cervical spondylosis / 中国骨伤

<p><b>OBJECTIVE</b>To compare the clinical effects and clinical indications between Mobi-C cervical artificial disc replacement (CADR) and MC+ anterior cervical decompression and fusion(ACDF) in treating cervical spondylosis.</p><p><b>METHODS</b>The clinical data of 100 patients with cervical spondylosis treated ACDF or CADR from June 2009 to June 2015 were retrospectively analyzed. There were 53 males and 47 females, aged from 38 to 70 years old. Among them, 50 cases were treated by ACDF (ACDF group), follow-up time was for 22 to 42 months with an average of (32.24±5.20) months;other 50 cases were treated by CADR (CADR group), follow-up time was for 23 to 48 months with an average of (30.40±5.66) months. Odom criterion was used to evaluate the clinical effects in two groups. JOA score, including sensory function, motor function and bladder function was used to assess the spinal cord function. Preoperative and postoperative responsible intervertebral space heights, cervical curvatures were compared by image data between two groups.</p><p><b>RESULTS</b>All incisions obtained good healing and no serious complications were found. At final follow-up, 30 cases got excellent results, 12 good, 8 fair in ACDF group;and 34 cases got excellent results, 10 good, 6 fair in CADR group;there was no significant difference between two groups(u=4.000, =0.827). At final follow-up, the scores of sensory function and motor function were obviously improved(<0.05), and bladder function had not obviously recovered (>0.05) in two groups;and CADR group in the scores of sensory function and motor function were obviously better than of ACDF group(<0.05). There was no significant difference in preoperative intervertebral space height, cervical curvature between two groups, and at final follow-up both had different recovered. The recovery of CADR group was obviously better than of ACDF group.</p><p><b>CONCLUSIONS</b>CADR can quickly recover normal action for patients and retains the movement. CADR has certain advantages in recovering cervical curvature, improveing sensory function and motor function, but it is not able to completely replace ACDF.</p>

A single-team experience with robotic pancreatic surgery in 1010 cases / 南方医科大学学报

<p><b>OBJECTIVE</b>To assess the safety and advantages of robotic pancreatic surgery (RPS) based on the single-team experience with 1010 cases.</p><p><b>METHODS</b>The clinical data of 1010 cases of RPS performed by a single team from November, 2011 to September, 2017 in our hospital were collected prospectively and analyzed. In most of cases the surgeries were performed using the third-generation da Vinci robotic surgical system.</p><p><b>RESULTS</b>The 1010 cases receiving RPS included 417 cases of robotic pancreatoduodenectomy (RPD), 428 cases of robotic distal pancreatectomy, 60 cases of robotic central pancreatectomy, 53 cases of robotic pancreatic tumor enucleation, 3 cases of Appleby procedure, and 49 cases of other operations (including 4 cases of innovative robotic retroperitoneal laparoscopic surgery, 4 cases of robotic pancreatic tumor enucleation combined with main pancreatic duct bridging repair, 1 case of single incision robotic pancreatic tumor enucleation, and 2 cases of robotic central pancreatectomy combined with end-to-end anastomosis reconstruction). The median operative time was 210 min (30-720 min) with a median intraoperative blood loss of 80 mL (10-2000 mL), a conversion rate of 4.06% (41/1010), a blood transfusion rate of 6.7% (68/1010), a mean post-operative stay of 10.87∓6.70 days, a complication rate (beyond grade III according to Clavien-Dindo scoring system) of 8.0% (81/1010), and a pancreatic fistula rate (beyond) grade B of 9.21% (93/1010). The mortality rate of the patients was 0.69% (7/1010) in 30 days and 1.31% (12//934) in 90 days. The application of RPS in total pancreatectomy increased steadily from the rate of 10.44% in 2012 to 72.06% in 2017.</p><p><b>CONCLUSION</b>This represents to our knowledge the world largest series of robotic pancreatic resections. RPS is expected to gradually replace open procedure and laparoscopic procedure to become the primary choice of approach for pancreatectomy. After the learning curve, RPS procedure including distal pancreatectomy, robotic Appleby procedure and other operations can be safely performed, and the experiences from other centers can be beneficial to reduce severe complications in the early stage of learning.</p>

Prognostic value of preoperative serum albumin in patients with non-muscle-invasive bladder cancer undergoing transurethral resection of bladder tumor / 南方医科大学学报

<p><b>OBJECTIVE</b>To assess the value of preoperative serum albumin level in predicting the survival of patients with non-muscle-invasive bladder cancer (NMIBC) undergoing transurethral resection of bladder tumor (TURBT).</p><p><b>METHODS</b>Two hundred and sixteen newly diagnosed patients with NMIBC who underwent TURBT between January, 2007 and April, 2012 were retrospectively analyzed. The patients were categorized into low albumin (<40 g/L) and normal albumin (≥40 g/L) groups. The patient survival was estimated using the Kaplan-Meier method, and univariate and multivariate Cox proportional analyses were used to determine the hazard ratios (HRs) for the overall survival (OS).</p><p><b>RESULTS</b>Of the patients with available data, 82 (39%) and 127 (61%) patients were classified into low albumin (<40 g/L) and normal albumin (≥40 g/L) groups, respectively. Kaplan-Meier analysis showed a significantly worse 5-year OS in low albumin group than in normal albumin group (P=0.017). In the multivariate Cox regression analysis, after adjusting for confounding variables, the preoperative albumin level remained as an independent predictor for 5-year OS (HR: 3.102, 95%CI: 1.200-8.020, P=0.020).</p><p><b>CONCLUSION</b>A low preoperative albumin level predicts a poor 5-year OS in patients with NMIBC who underwent TURBT. Preoperative serum albumin can be a good prognostic factor for predicting survival of the patients with NMIBC treated with TURBT.</p>

Reproduction,genotype identification and evaluation of APP/PS1 transgenic mice / 中国应用生理学杂志

OBJECTIVES@#To identify the genotype of (APP/PS1) transgenic mice and evaluate the changing of cognitive and behavioral fu nctions, provide an effective animal model for the Alzheimer's disease (AD) research.@*METHODS@#Male APP/PS1 transgenic mice mated with female APP/PS1 transgenic mice, and the genotype of their filial mice was identified by PCR. The APP +/PS1 + mice were assigned into AD model group (AD group, =8), and the APP/PS1 mice were assigned into control group (CT group, =8). The Morris water maze test was carried out to detect the capacity of learning and memory of mice. After that, the mice were sacrificed and the brain tissues were sampled and stained by HE and congo red for the pathological examination.@*RESULTS@#①A APP/PS1 genome DNA about 360 bp size was detected. The methods of feeding and breeding were successful to attain APP/PS1 transgenic mice.②Statistical significance was found in the differences of the capacity of learning and memory between 7-month-old APP/PS1 positive mice and negative mice (<0.05).③The results of HE stain showed that the structure and cellular morphology of hippocampus of AD mice were obviously abnormal. The results of congo red stain showed that positive amyloid plaque was observed in brains of AD mice.@*CONCLUSIONS@#APP/PS1 transgenic mice present typical symptoms and behaviors of Alzheimer's disease. The transgenic mouse is an effective tool for the research and prevention of AD.

Expression of hypoxia inducible factor-2 alpha in overloaded- stress induced destruction of mandibular condylar chondrocytes.

OBJECTIVE: To study the protein expression of HIF-2α in condylar chondrocytes under the different stress loading, to investigate the possible effects of HIF-2α involved in the mortality of condylar chondrocytes under overloaded- stress. MATERIALS AND METHODS: Chondrocytes were isolated from TMJ condylar cartilage and cultured in hypoxia-incubator. Chondrocytes were divided into 4 groups: 0, 1000, 2000, 3000 ustrain group, which was subjected to cyclic tensile strain (CTS) of 0.5Hz for 2h. The rate of cell mortality was calculated. Western blot was used to measure the expression of HIF-2α and it's downstream catabolic factors (MMP3, MMP13, ADAMTS4) in protein levels respectively. RESULTS: With the increase of CTS, both of the rate of cell mortality and protein expression of HIF-2α increased significantly (p<0.05). The same tendency was also found in it's downstream catabolic factors (MMP3, MMP13, ADAMTS4) in protein levels (p<0.05). CONCLUSIONS: The results indicated that elevated expression of HIF-2α may be a possible mechanism related to overloaded- stress induced mortality of condylar chondrocytes.