OBJECTIVES: To investigate the expression levels of marginal zone B and B1-cell-specific protein (MZB1) in pan-cancer and its association with patient prognosis and tumor microenvironment (TME). METHODS: MZB1 expression data, clinicopathological parameters, and survival data from 33 cancer types were extracted from the UCSC database for analyzing the correlations of MZB1 with clinical stage, patient prognosis, immunomodulatory genes, immune checkpoint genes, tumor stemness, immune cell infiltration, tumor mutational burden (TMB), and microsatellite instability (MSI). MZB1 gene mutations in pan-cancer were assessed using cBioPortal online database, and the value of MZB1 for cancer diagnosis was evaluated using ROC curve analysis. MZB1 expression levels in myeloid leukemia and renal carcinoma cells were detected using RT-qPCR and Western blotting, and the effect of MZB1 knockdown on cell proliferation was examined using EdU assay. RESULTS: MZB1 was significantly overexpressed in 20 cancer types, including kidney renal clear cell carcinoma (KIRC), breast invasive carcinoma, and acute myeloid leukemia. Its expression was associated with TNM stage, clinical stage, overall survival, and progression-free survival in multiple cancers. In most tumors, MZB1 expression was correlated significantly with immunomodulatory genes, immune checkpoint genes, tumor stemness, immune cell infiltration, TMB, and microsatellite instability. Gene amplification was the predominant mutation type of MZB1 in pan-cancer, and MZB1 showed high diagnostic value for skin cutaneous melanoma, KIRC, and head and neck squamous cell carcinoma. MZB1 was highly expressed in different myeloid leukemia cell lines and renal carcinoma cell lines, and MZB1 knockdown significantly suppressed the proliferation of HL60 and 769-P cells. CONCLUSIONS MZB1 is highly expressed in a variety of tumors, and its aberrant expression affects the occurrence and prognosis of many tumors, suggesting its potential as a novel tumor biomarker and immunomodulatory target.
Humans ; Prognosis ; Tumor Microenvironment ; Neoplasms/pathology* ; Cell Line, Tumor ; Mutation ; Kidney Neoplasms ; Microsatellite Instability ; Cell Proliferation ; Carcinoma, Renal Cell

The comparison between traditional Chinese medicine Jinzhen oral liquid (JZOL) and Western medicine in treating children with acute bronchitis (AB) showed encouraging outcomes. This trial evaluated the efficacy and safety of the JZOL for improving cough and expectoration in children with AB. 480 children were randomly assigned to take JZOL or ambroxol hydrochloride and clenbuterol hydrochloride oral solution for 7 days. The primary outcome was time-to-cough resolution. The median time-to-cough resolution in both groups was 5.0 days and the antitussive onset median time was only 1 day. This randomized controlled trial showed that JZOL was not inferior to cough suppressant and phlegm resolving western medicine in treating cough and sputum and could comprehensively treat respiratory and systemic discomfort symptoms. Combined with clinical trials, the mechanism of JZOL against AB was uncovered by network target analysis, it was found that the pathways in TRP channels like IL-1β/IL1R/TRPV1/TRPA1, NGF/TrkA/TRPV1/TRPA1, and PGE2/EP/PKA/TRPV1/TRPA1 might play important roles. Animal experiments further confirmed that inflammation and the immune regulatory effect of JZOL in the treatment of AB were of vital importance and TRP channels were the key mechanism of action.

The global COVID-19 coronavirus pandemic has infected over 109 million people, leading to over 2 million deaths up to date and still lacking of effective drugs for patient treatment. Here, we screened about 1.8 million small molecules against the main protease (M^pro) and papain like protease (PL^pro), two major proteases in severe acute respiratory syndrome-coronavirus 2 genome, and identified 1851M^pro inhibitors and 205 PL^pro inhibitors with low nmol/l activity of the best hits. Among these inhibitors, eight small molecules showed dual inhibition effects on both M^pro and PL^pro, exhibiting potential as better candidates for COVID-19 treatment. The best inhibitors of each protease were tested in antiviral assay, with over 40% of M^pro inhibitors and over 20% of PL^pro inhibitors showing high potency in viral inhibition with low cytotoxicity. The X-ray crystal structure of SARS-CoV-2 M^pro in complex with its potent inhibitor 4a was determined at 1.8 Å resolution. Together with docking assays, our results provide a comprehensive resource for future research on anti-SARS-CoV-2 drug development.
Humans ; Antiviral Agents/chemistry* ; COVID-19 ; COVID-19 Drug Treatment ; High-Throughput Screening Assays ; Molecular Docking Simulation ; Protease Inhibitors/chemistry* ; SARS-CoV-2/enzymology* ; Viral Nonstructural Proteins

Animals ; Monkeypox/drug therapy* ; Antiviral Agents/therapeutic use* ; Phosphoric Monoester Hydrolases/therapeutic use* ; Macaca fascicularis

The application of clustered regularly interspaced short palindromic repeats (CRISPR) and CRISPR-associated proteins (Cas) can be limited due to a lack of compatible protospacer adjacent motif (PAM) sequences in the DNA regions of interest. Recently, SpRY, a variant of Streptococcus pyogenes Cas9 (SpCas9), was reported, which nearly completely fulfils the PAM requirement. Meanwhile, PAMs for SpRY have not been well addressed. In our previous study, we developed the PAM Definition by Observable Sequence Excision (PAM-DOSE) and green fluorescent protein (GFP)‍-reporter systems to study PAMs in human cells. Herein, we endeavored to identify the PAMs of SpRY with these two methods. The results indicated that 5'-NRN-3', 5'-NTA-3', and 5'-NCK-3' could be considered as canonical PAMs. 5'-NCA-3' and 5'-NTK-3' may serve as non-priority PAMs. At the same time, PAM of 5'-NYC-3' is not recommended for human cells. These findings provide further insights into the application of SpRY for human genome editing.
CRISPR-Associated Protein 9/metabolism* ; CRISPR-Cas Systems ; DNA ; Gene Editing/methods* ; Humans ; Streptococcus pyogenes/metabolism*

The demand for rehabilitation therapists is rising in response to social development, disease spectrum changes and population aging. Under the guidance of the integration of rehabilitation and clinical medicine, Beijing Rehabilitation Hospital has carried out a comprehensive scientific design and practice in the sub-professional training mode for new rehabilitation therapists according to the discipline development and clinical needs, strengthened their training of sub-professional skills, and provided an effective way to standardize the profession admission and specialty advancement.

Objective: To explore relationships between the enrichment of ETBF, Fn, Hp in feces, tissues and colorectal cancer. Methods: Feces, lesion tissue and adjacent tissue from 24 patients with colorectal cancer and 31 patients with adenomas were collected, and we collected Feces and tissue of 20 healthy control persons. Then the copy numbers of enterotoxigenic B. fragilis (ETBF), Fusobacterium nucleatum (Fn) and Helicobacter pylori (Hp) were determined by quantitative real-time PCR. Immunohistochemical method was used to examine the expression intensity of EGFR and p53, and the relationships between different expression intensity of EGFR, p53 and the numbers of three bacterias. Results: In the feces, copy numbers of ETBF and Fn were as follous: colorectal cancer group>adenomas group>healthy control group (P<0.05). Copy numbers of Hp were as follous: colorectal cancer group>healthy control group (P<0.01); adenomas group>healthy control group (P<0.01). In the tissue, copy numbers of ETBF, Fn were as follows: colorectal cancer group>adenomas group>healthy control group (P<0.05). Copy numbers of Hp were as follows: colorectal cancer group>healthy control group (P<0.01); adenomas group>healthy control group (P<0.01). Copy numbers of those three bacteria in the lesion tissue and the adjacent tissue had no significant difference. This happened both in colorectal cancer group and adenomas group. The different expression intensity of EGFR, p53 and the number of three bacteria showed no obviously statistical correlation(P>0.05). Conclusion Adenomatous polyp and colorectal cancer patients show high enrichment of ETBF, Fn and Hp in both feces and tissues. ETBF, Fn and Hp probably contribute to the development of adenomatous polyp and colorectal cancer. Trial registration Chinese Clinical Trial Registry, ChiCTR-BOC-17012509.

Objective: To investigate the changes of the intestinal mucosa-associated microbiota in the patients with ulcerative colitis (UC), and to explore their correlation with the clinical manifestations. Methods: From June to October 2016, at Gastrointestinal Endoscopy Center, the Second Affiliated Hospital of Xi′an Jiaotong University, 28 patients with UC and 16 healthy individuals who underwent colonoscopy examination were enrolled. The mucosa specimens of them were collected for fluorescent in situ hybridization (FISH). The bacterial flora were observed and counted, the correlation between the bacterial flora and the clinical manifestations were analyzed. Kruskal-Wallis test and Spearman rank correlation analysis were performed for statistical analysis. Results: Among 28 patients with UC, 16 were at active phase and 12 at remission phase. The number of total bacteria flora, Escherichia coli, Clostridium and Bacteroides of the active UC group and remission UC group were all more than those of healthy control group; however the number of Lactobacillus and Bifidobacteria were less than those of healthy control group, and the differences were statistically significant (χ²=23.34, 19.94; 23.40, 12.96; 23.39, 19.16; 23.32, 10.46; 23.19, 4.25; 18.94, 12.33; all P<0.05). The number of total bacteria flora, Escherichia coli and Bacteroides of the active UC group were more than those of the remission UC group, however the number of Lactobacillus and Bifidobacteria were less than those of the remission UC group, and the differences were statistically significant (χ²=7.32, 5.63, 5.62, 20.38 and 4.82; all P<0.05). In the patients with UC, the defecation frequency was positively correlated with the count of Bacteroides (r=0.459, P=0.014) and was negatively correlated with the count of Lactobacillus (r=-0.634, P<0.01). In UC patients, bloody stool, endoscopic appearance and total Mayo score were positively correlated with the counts of universal bacteria, Escherichia coli and Bacteroides (r=0.469, 0.403, 0.376; 0.604, 0.562, 0.475; 0.551, 0.463, 0.461; all P <0.05); and which were negatively correlated with Lactobacillus and Bifidobacteria (r=-0.570, -0.413; -0.899, -0.458; -0.862, -0.480; all P<0.05). The evaluation by the doctors was positively correlated with the counts of Escherichia coli (r=0.415, P=0.028), however was negatively correlated with Lactobacillus and Bifidobacteri (r=-0.841, -0.529; both P<0.01). Conclusion The intestinal microbiota have sigificantly changed in patients with UC which are correlated with some clinical manifestations of UC.

Objective To investigate the effects of p38 MAPK inhibitor on glutamate (Glu), glutamine (Gln), taurine amino acid (Tau), glycine (Gly) and gamma aminobutyric acid (GABA) in hippocampus of rats with vascular dementia (VD).Methods Twenty-four healthy male SD rats were randomly divided into sham operation group, VD model group and inhibitor group. VD model was established by permanent ligation of bilateral carotid artery method, and the sham operation group stripping bilateral carotid artery but not ligation. Rats of the inhibitor group were injected p38 MAPK inhibitor SB202190 after the establishment of VD model. Morris water maze was used to evaluate the learning and memory function of rats. The samples of DG region of hippocampus were collected by microdialysis, and the contents of amino acids were detected. Results The learning and memory abilities were significantly better in sham operation group and p38 MAPK inhibitor group than those of VD model group. The latency time was significantly shorter in p38 MAPK group than that of VD model group. The time of locating the platform quadrant and the number of crossing the original platform were significantly higher in sham operation group and p38 MAPK inhibitor group than those of VD model group (P <0.05). Compared with sham group, the levels of Glu, Gln and Tau were significantly lower, and Gly and GABA were significantly higher, in VD group and p38 MAPK inhibitor group (P<0.05). Compared with VD group, Glu, Gln and Tau were significantly increased, Gly and GABA were significantly decreased in p38 MAPK inhibitor group (P<0.05). Conclusion p38 MAPK inhibitor has protective effect on VD hippocampal injury, which may be related to its ability to inhibit VD-induced abnormal secretion of amino acids and regulate the secretion of various amino acids.

BACKGROUND:Transplantation of alogeneic intervertebral disc can be facilitated by the cryopreservation of the intervertebral disc. But the traditional cryopreservation methods always lead to the appearing of ice crystals inside and outside the cels which can cause celular injury. The vitrification method that can avoid the formation of ice crystals have been widely applied in the cryopreservation field. However, only a few reports have assessed the vitrified cryopreservation of the intervertebral disc, and the toxicity of cryoprotectants to the nucleus pulposus cels have not been fuly explored.
OBJECTIVE:To determine the order of toxicity of five commonly used cryoprotectants that are used alone or in combination to rabbit nucleus pulposus cels, and to select the optimal cryoprotectant for the vitrification of the intervertebral disc.
METHODS: We chose five most commonly used cryoprotectants including dimethyl sulphoxide, formamide, ethylene glycol, propylene glycol and glycerol. Then, 5 single commonly used cryoprotectants, 10 mixed agents containing any 2 commonly used cryoprotectants, and 10 mixed agents containing any 3 commonly used cryoprotectants were formulated. Cel viability of nucleus pulposus cels was determined using cel counting kit-8 and fluorescein diacetate/propidium iodide method. Al data obtained were analyzed statisticaly to choose the appropriate combining scheme with less toxicity.
RESULTS AND CONCLUSION: The order of the toxicity of these five commonly used cryoprotectants from low to high was ethylene glycol, glycerol, formamide, dimethyl sulphoxide, and propylene glycol. The toxicity of the combined agents containing two or three commonly used cryoprotectants was lower than that of any commonly used cryoprotectants that were used to formulate them. The toxicity of the mixed agents that contained ethylene glycol or glycerol was lower than that of any other mixed agents. So we can choose the mixed cryoprotectants that contain ethylene glycol and (or) glycerol for the vitrification of the intervertebral disc.