This study aims to investigate the protective effects and mechanisms of polysaccharide extract PCP1 from Polygonatum cyrtonema in ameliorating cerebral ischemia-reperfusion(I/R) injury in rats through modulation of the Toll-like receptor 4(TLR4)/NOD-like receptor protein 3(NLRP3) signaling pathway. In vivo, SD rats were randomly divided into the sham group, model group, PCP1 group, nimodipine(NMDP) group, and TLR4 signaling inhibitor(TAK-242) group. A middle cerebral artery occlusion/reperfusion(MCAO/R) model was established, and neurological deficit scores and infarct size were evaluated 24 hours after reperfusion. Hematoxylin-eosin(HE) and Nissl staining were used to observe pathological changes in ischemic brain tissue. Transmission electron microscopy(TEM) assessed ultrastructural damage in cortical neurons. Enzyme-linked immunosorbent assay(ELISA) was used to measure the levels of interleukin-1β(IL-1β), interleukin-6(IL-6), interleukin-18(IL-18), tumor necrosis factor-α(TNF-α), interleukin-10(IL-10), and nitric oxide(NO) in serum. Immunofluorescence was used to analyze the expression of TLR4 and NLRP3 proteins. In vitro, a BV2 microglial cell oxygen-glucose deprivation/reperfusion(OGD/R) model was established, and cells were divided into the control, OGD/R, PCP1, TAK-242, and PCP1 + TLR4 activator lipopolysaccharide(LPS) groups. The CCK-8 assay evaluated BV2 cell viability, and ELISA determined NO release. Western blot was used to analyze the expression of TLR4, NLRP3, and downstream pathway-related proteins. The results indicated that, compared with the model group, PCP1 significantly reduced neurological deficit scores, infarct size, ischemic tissue pathology, cortical cell damage, and the levels of inflammatory factors IL-1β, IL-6, IL-18, TNF-α, and NO(P<0.01). It also elevated IL-10 levels(P<0.01) and decreased the expression of TLR4 and NLRP3 proteins(P<0.05, P<0.01). Moreover, in vitro results showed that, compared with the OGD/R group, PCP1 significantly improved BV2 cell viability(P<0.05, P<0.01), reduced cell NO levels induced by OGD/R(P<0.01), and inhibited the expression of TLR4-related inflammatory pathway proteins, including TLR4, myeloid differentiation factor 88(MyD88), tumor necrosis factor receptor-associated factor 6(TRAF6), phosphorylated nuclear factor-kappaB dimer RelA(p-p65)/nuclear factor-kappaB dimer RelA(p65), NLRP3, cleaved-caspase-1, apoptosis-associated speck-like protein(ASC), GSDMD-N, IL-1β, and IL-18(P<0.05, P<0.01). The protective effects of PCP1 were reversed by LPS stimulation. In conclusion, PCP1 ameliorates cerebral I/R injury by modulating the TLR4/NLRP3 signaling pathway, exerting anti-inflammatory and anti-pyroptotic effects.
Animals ; Toll-Like Receptor 4/genetics* ; NLR Family, Pyrin Domain-Containing 3 Protein/genetics* ; Rats, Sprague-Dawley ; Rats ; Reperfusion Injury/genetics* ; Male ; Signal Transduction/drug effects* ; Polysaccharides/isolation & purification* ; Polygonatum/chemistry* ; Brain Ischemia/genetics* ; Drugs, Chinese Herbal/administration & dosage* ; Mice ; Humans

OBJECTIVE: To establish and modify quantitative real-time polymerase chain reaction (qPCR)-based serotyping assays to distinguish 97 pneumococcal serotypes. METHODS: A database of capsular polysaccharide ( cps) loci sequences was generated, covering 97 pneumococcal serotypes. Bioinformatics analyses were performed to identify the cps loci structure and target genes related to different pneumococcal serotypes with specific SNPs. A total of 27 novel qPCR serotyping assay primers and probes were established based on qPCR, while 27 recombinant plasmids containing serotype-specific DNA sequence fragments were constructed as reference target sequences to examine the specificity and sensitivity of the qPCR assay. A panel of pneumococcal reference strains was employed to evaluate the capability of pneumococcal serotyping. RESULTS: A total of 97 pneumococcal serotyping assays based on qPCR were established and modified, which included 64 serotypes previously reported as well as an additional 33 serotypes. Twenty-seven novel qPCR serotyping target sequences were implemented in the pneumococcal qPCR serotyping system. A total of 97 pneumococcal serotypes, which included 52 individual serotypes and 45 serotypes belonging to 20 serogroups, could not be identified as individual serotypes. The sensitivity of qPCR assays based on 27 target sequences was 1-100 copies/µL. The specificity of the qPCR assays was 100%, which were tested by a panel of 90 serotypes of the pneumococcal reference strains. CONCLUSION A total of 27 novel qPCR assays were established and modified to analyze 97 pneumococcal serotypes.
Real-Time Polymerase Chain Reaction ; Serotyping ; Streptococcus pneumoniae/genetics* ; Serogroup

Objective: To evaluate the clinical value of the MeltPro MTB assays in the diagnosis of drug-resistant tuberculosis. Methods: A cross-sectional study design was used to retrospectively collect all 4 551 patients with confirmed tuberculosis between January 2018 and December 2019 at Beijing Chest Hospital, Capital Medical University. Phenotypic drug sensitivity test and GeneXpert MTB/RIF (hereafter referred to as "Xpert") assay were used as gold standards to analyze the accuracy of the probe melting curve method. The clinical value of this technique was also evaluated as a complementary method to conventional assays of drug resistance to increase the detective rate of drug-resistant tuberculosis. Results: By taking the phenotypic drug susceptibility test as the gold standard, the sensitivity of the MeltPro MTB assays to detect resistance to rifampicin, isoniazid, ethambutol and fluoroquinolone was 14/15, 95.7%(22/23), 2/4 and 8/9,respectively; and the specificity was 92.0%(115/125), 93.2%(109/117), 90.4%(123/136) and 93.9%(123/131),respectively; the overall concordance rate was 92.1%(95%CI:89.6%-94.1%),and the Kappa value of the consistency test was 0.63(95%CI:0.55-0.72).By taking the Xpert test results as the reference, the sensitivity of this technology to the detection of rifampicin resistance was 93.6%(44/47), the specificity was100%(310/310), the concordance rate was 99.2%(95%CI:97.6%-99.7%), and the Kappa value of the consistency test was 0.96(95%CI:0.93-0.99). The MeltPro MTB assays had been used in 4 551 confirmed patients; the proportion of patients who obtained effective drug resistance results increased from 83.3% to 87.8%(P<0.01); and detection rate of rifampicin, isoniazid, ethambutol, fluoroquinolone resistance, multidrug and pre-extensive drug resistance cases were increased by 3.2%, 14.7%, 22.2%, 13.7%, 11.2% and 12.5%, respectively. Conclusion: The MeltPro MTB assays show satisfactory accuracy in the diagnosis of drug-resistant tuberculosis. This molecular pathological test is an effective complementary method in improving test positivity of drug-resistant tuberculosis.
Humans ; Rifampin/therapeutic use* ; Antibiotics, Antitubercular/therapeutic use* ; Mycobacterium tuberculosis ; Ethambutol/pharmacology* ; Isoniazid/pharmacology* ; Paraffin Embedding ; Retrospective Studies ; Cross-Sectional Studies ; Drug Resistance, Bacterial ; Sensitivity and Specificity ; Tuberculosis, Multidrug-Resistant/drug therapy*

Objective:To explore the expression of calcium-binding protein 39 (CAB39) and vascular endothelial growth factor (VEGF) in bladder cancer tissues and their relationship with the prognosis of patients.Methods:A case-control study was conducted to select the focal tissues (case group) and post-operative paracancourous tissues (control group) from 94 bladder cancer patients who underwent surgery from Zibo Central Hospital from June 2014 to April 2018. Western blot and immunohistochemical staining were used to detect the expression of VEGF and CAB39 proteins in the two specimens. The positive expression rates of VEGF and CAB39 protein in bladder cancer tissues with different pathological characteristics were analyzed, and the relationship between VEGF and CAB39 protein expression and prognosis of patients with bladder cancer was analyzed by Cox proportional hazard regression model.Results:The relative expression levels of VEGF and CAB39 protein in the case group were significantly higher than those in the control group, with significant statistical difference (all P<0.05). The immunohistochemical staining scores of VEGF and CAB39 protein in the case group were higher than those in the control group, with statistically significant difference (all P<0.05). The positive expression rate of CAB39 protein was significantly higher in bladder cancer tissure of patients with low-grade, clinical stage T3-T4 and lymph node metastasis, with statistically significant difference (all P<0.05). The positive expression rate of VEGF protein was significantly higher in bladder cancer tissue of patients with clinical stage T3-T4 and lymph node metastasis, with statistically significant difference (all P<0.05). Cox proportional hazard regression model showed that low grade, clinical stage T3-T4, lymph node metastasis, positive expression of VEGF and CAB39 protein were independent risk factors for poor prognosis in bladder cancer patients in 3 years (all P<0.05). Conclusions:VEGF and CAB39 proteins are highly expressed in bladder cancer tissue, and have a certain relationship with the poor prognosis of patients.

OBJECTIVES: To investigate the clinical effect of the arytenoid cartilage reposition using snake mouth reduction forceps under general anesthesia. METHODS: Data of twenty-six cases accepted arytenoid cartilage reposition under intravenous general anesthesia were analyzed, nineteen cases accepted laryngeal CT scan and cricoarytenoid joint reconstruction, all patients underwent endolaryngeal muscle electromyography examination. According to the position of cartilage dislocation prompted by laryngoscope and CT, the arytenoid cartilage was repositoned under the visual laryngoscope using special snake mouth reduction forceps. If bilateral arytenoid cartilage were still asymmetrically at the end of the surgery, patients needed repeated reposition 1 to 2 times 1 week after operation. The efficacy was evaluated 4 weeks later. RESULTS: All patients had a hoarse and breathing voice preoperative. Under laryngoscope, there were different degrees of vocal cord movement disorders accompanied by incomplete glottis closure, 22 cases happened in left side and 4 in right side. The arytenoid cartilage was dislocated anteromedially in 25 cases and posterolaterally in 1 case. CT showed that 15 cases of arytenoid cartilage were tilted anteromedially; the interval of the cricoarytenoid joint was widened. In axial CT images, there were no direct signs of the arytenoid cartilage dislocation in the 4 cases, but the abnormal position was seen in the reconstruction images. The laryngeal electromyography indicated that 7 cases were abnormal, duration of motor unit potential were visible and the raising potential were mixed. There were 4 patients with normal voice in the first day after surgery, and 19 cases underwent twice and 3 cases underwent three times surgery. Vioce became normal in 4 weeks. Swallowing pain and bucking were all disappeared. Vocal cords movement were recovered to normal level in 25 cases. In 1 case with neck strangulation, the vocal cord movement was slightly worse than health side, but significantly better than that before operation. CONCLUSIONS The arytenoid cartilage reposition using snake mouth reduction forceps under general anesthesia was an effective method for the treatment of the cricoary-tenoid joint dislocation.
Anesthesia, General ; instrumentation ; Arytenoid Cartilage ; injuries ; Hoarseness ; Humans ; Laryngoscopes ; Mouth ; Surgical Instruments

Objective To investigate the role of metastatic lymph node ratio (MLR) in the evaluation of prognosis of patients with gastric cancer (GC) at different lymph node numbers examined.Methods Clinical data were reviewed retrospectively in a total 535 patients who underwent surgery for GC.Spearman correlation analysis between MLR or number of metastatic lymph nodes (N) and examined lymph node numbers,Kaplan-Meier method was used for comparison survival rates of N stage and MLR stage.A receiver operating characteristic (ROC) curve was used to evaluate the role of N stage and MLR stage in the prognosis of GC patients.Results Metastatic lymph node ratio and number of metastatic lymph nodes correlated with the examined lymph node numbers (r =0.146,r =0.378,P ＜ 0.01,P ＜ 0.001).The 5 year survival rate of MLR0,MLR1,MLR2 and MLR3 patients were 57.5％,69.9％,40.0％ and 21.7％ respectively when examined lymph node numbers ＜ 6 (P ＜ 0.01).The 5-YSR of MLR0,MLR1,MLR2 and MLR3 patients were 86.8％,59.2％,35.8％ and 39.2％ respectively when between 6-10 (P ＜0.001) and the 5-YSR of MLR0,MLR1,MLR2 and MLR3 patients were 88.7％,62.5％,0 and 17.7％ respectively when they ＞ 10(P ＜0.001).AUC of MLR staging was 0.68 ±0.05 when the numbers ＜ 6 (P ＜ 0.001).AUC of MLR staging was 0.72 ± 0.04 at numbers 6-10 (P ＜ 0.001).AUC of MLR staging was 0.79 ± 0.03 when numbers ＞ 10 (P ＜ 0.001).Conclusions MLR was less influenced by lymph node number examined than N.MLR stage has potential superiority to that the N stage in assessing prognosis of GC patients,especially for patients with more than 6 lymph nodes examined.

OBJECTIVEThe influence of anti-tuberculosis (TB) treatment history on tuberculous lymphadenitis (TBLN) diagnosis is unclear. Therefore, this study aims to evaluate the diagnostic methods, including histology, microbiology, and molecular tests, used for TBLN.

METHODSIn this study, suspected patients with TBLN and having different anti-TB treatment background were enrolled. All the samples were tested simultaneously by histology, Ziehl-Neelsen (ZN) staining, mycobacterial culture (culture), Xpert MTB/RIF (xpert), real-time PCR, and high-resolution melting curve PCR (HRM). Thereafter, the performance of these methods on samples with different anti-TB treatment background was assessed.

RESULTSIn our study, 89 patients were prospectively included 82 patients with TBLN and 7 with other diseases. The overall sensitivities of Xpert, real-time PCR, histology, ZN staining, and culture were 86.6%, 69.5%, 58.5%, 43.9%, and 22.0%, respectively. The anti-TB treatment history revealed dramatic influences on the sensitivity of culture (P < 0.0001). In fact, the treatment that lasted over 3 months also influenced the sensitivity of Xpert (P < 0.05). However, the treatment history did not affect the performance of remaining tests (P > 0.05). For rifampicin drug susceptibility test (DST), the anti-TB treatment showed only significant influence on the success rate of culture DST (P = 0.001), but not on those of Xpert and HRM tests (P > 0.05).

CONCLUSIONOther tests as well as culture should be considered for patients with TBLN having retreatment history or over 1-month treatment to avoid false negative results.

Adolescent ; Adult ; Aged ; Antitubercular Agents ; therapeutic use ; Bacteriological Techniques ; Drug Resistance, Bacterial ; Female ; Humans ; Male ; Middle Aged ; Tuberculosis, Lymph Node ; diagnosis ; drug therapy ; microbiology ; Young Adult

OBJECTIVETo assess the beneficial and adverse effects of Wendan Decoction (温胆汤, WDD) for the treatment of schizophrenia.

METHODSFive electronic databases were searched until May 2014, including the Chinese National Knowledge Infrastructure, the Chinese Biomedical Literature Database, the Chinese Scientist Journal Database, PubMed, and the Cochrane Central Register of Controlled Trials in the Cochrane Library. The randomized controlled trials (RCTs) testing WDD against placebo, antipsychotic drugs, or WDD combined with antipsychotic drugs against antipsychotic drugs alone were included. Study selection, data extraction, quality assessment, and data analyses were conducted according to the Cochrane standards.

RESULTSThirteen RCTs (involving 1,174 patients) were included and the methodological quality was evaluated as generally low. The pooled results showed that WDD combined with antipsychotic drugs were more effective in clinical comprehensive effect, Positive and Negative Syndrome Scale (PANSS) scores and Brief Psychiatric Rating Scale scores compared with antipsychotic drugs alone. However, WDD had less effectiveness compared with antipsychotics in clinical comprehensive effect; and WDD was not different from antipsychotic drugs for PANSS scores. The side effects were significantly reduced in the intervention group compared with the control group.

CONCLUSIONSWDD appears to be effective on improving symptoms in patients with schizophrenia. However, due to poor methodological quality in the majority of the included trials, the potential benefit from WDD needs to be confirmed in rigorous trials and the design and reporting of trials should follow the international standards.

Antipsychotic Agents ; therapeutic use ; Brief Psychiatric Rating Scale ; Drugs, Chinese Herbal ; therapeutic use ; Humans ; Publication Bias ; Randomized Controlled Trials as Topic ; Schizophrenia ; drug therapy

OBJECTIVETo investigate the effect of Emodin combined with 3'-azido-3'-deoxythymidine (AZT) on the proliferation and apoptosis of concentrated leukemia stem cells (CLSC)-human acute myeloid leukemia KG-la cells and expression of BCL-2, NF-κB and TGF-β.

METHODSThe tumor stem cell-like subpopulation in human leukemia cell line KG-1a was enriched with 5-fluorouracil (5-FU). The CD34⁺ CD38⁻ subpopulation in the KG-1a cells was detected with flow cytometry, the cell proliferation was detected by MTT method to study the of Emodin and AZT in the CLSC. The cell apoptosis was analyzed by flow cytometry. The expression of NF-κB, BCL-2 and TGF-β mRNA and proteins were measured with RT-PCR and Western blot respectively.

RESULTSAs compared with cells treated with mentioned above drugs alone, the inhibition of proliferation potential and apoptosis rate of cells in combination group markedly increase with time and concentration dependent member (P < 0.01), the expression of NF-κB, BCL-2 and TGF-β mRNA and proteins decreased.

CONCLUSIONEmodin combined AZT can synergistically inhibit the proliferation, induce cell apoptosis, and down regulate the expression of NF-κB, BCL-2 and TGF-β mRNA and proteins in the CLSC, the possible mechanism of synergistic effect may be associated with inhibiton of BCL-2 activation and down-regulation of the expression of NF-κB, and TGF-β.

Apoptosis ; Cell Line, Tumor ; Cell Proliferation ; Down-Regulation ; Emodin ; pharmacology ; Humans ; Leukemia ; NF-kappa B p50 Subunit ; metabolism ; Neoplastic Stem Cells ; cytology ; drug effects ; metabolism ; Proto-Oncogene Proteins c-bcl-2 ; metabolism ; Transforming Growth Factor beta1 ; metabolism ; Zidovudine ; pharmacology