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Objective: Differential flora and differential metabolites shared by the intestinal and respiratory tracts of rats were screened to analyze the possible role of changes in intestinal flora and metabolites in the progression of pneumoconiosis in rats. Methods: In April 2020, 18 SD rats were randomly divided into three groups (control group, coal mine dust group and silica group, 6 in each group) , rats in the coal mine dust group and silica group were perfused with 1 ml of 50 mg/ml coal mine well dust suspension and silica suspension by nontracheal exposure, respectively. While rats in the control group were perfused with an equal dose of sterilized normal saline. Twenty four weeks after dust staining, rat feces, throat swabs, and lung lavages were collected. 16SrDNA gene sequencing and UHPLC-QTOF-MS untargeted metabolomics were used to analyze the flora and metabolites in feces, throat swabs and lung lavage fluid of rats in each group, to screen for shared differential flora and shared differential metabolites in intestinal and respiratory tract, and the correlation analysis between the differential flora and metabolites was performed using Spearman's statistics. Results: Compared with the control group, a total of 9 species shared differential flora between intestinal and respiratory tract were screened at phylum level, and a total of 9 species shared differential genus between intestinal and respiratory tract were screened at genus level in the coal mine dust group, mainly Firmicutes, Actinobacteria, Streptococcus, Lactobacillus, etc. Compared with the control group, a total of 9 shared differential flora were screened at the phylum level, and a total of 5 shared differential genus were screened at the genus level in the silica group, mainly Proteobacteria, Actinobacteria, Allobactera, Mucilaginibacter, etc. Compared with the control group, a total of 7 shared differential metabolites were screened for up-regulation of Stigmatellin, Linalool oxide and Isoleucine-leucine in both intestinal and respiratory tract in the coal mine dust group. Compared with the control group , a total of 19 shared differential metabolites werescreened in the silica group, of which Diethanolamine, 1-Aminocyclopropanecarboxylic acid, Isoleucine-leucine, Sphingosine, Palmitic acid, D-sphinganine, 1, 2-dioleoyl-sn-glycero-3-phosphatidylcholine, and 1-Stearoyl-2-oleoyl-sn-glycerol 3-phosphocholine were up-regulated in both the intestinal and respiratory tract. Conclusion: There is a translocation of intestinal and respiratory flora in pneumoconiosis rats, and rats have an imbalance of lipid metabolism during the progression of pneumoconiosis.
Subject(s)
Rats , Animals , Isoleucine , Leucine , Coal Mining , Rats, Sprague-Dawley , Pneumoconiosis , Dust/analysis , Silicon Dioxide , CoalABSTRACT
Objective: To collect, compare and analyze the changes in the strength and stability of the forearm Rou-Kneading manipulation before and after the training in Shaolin Neigong (internal Qigong). Methods: Ninety first-year undergraduates were randomized into three groups using the random number table method, with 30 people in each group. The Gongfa (Qigong method) group received training in both Shaolin Neigong and forearm Rou-Kneading manipulation. The manipulation group only received training in forearm Rou-Kneading manipulation. The control group only received training in forearm Rou-Kneading manipulation for one week. The ZTC-1 intelligent Tuina (Chinese therapeutic massage) manipulation parameter detection system was used to collect the wave crest, wave trough, and crest-trough difference of the strength and frequency of the forearm Rou-Kneading manipulation on the Z-axis (up and down), X-axis (left and right) and Y-axis (backward and forward) at weeks one, five and ten from the three groups. The collected data were then processed and analyzed. Results: The intra-group comparisons showed statistical significance in the Gongfa group and manipulation group (P<0.05). The strength and stability shown on the axes Z, X, and Y constantly grew with the increase of training time in the Gongfa group. The wave crest on the axes Z and Y steadily rose in the manipulation group, as well as the frequency on the axes Z, X and Y. The control group failed to show statistical significance in any of the three times of intra-group comparisons (P>0.05). The between-group comparisons showed statistical significance among the three groups at weeks five and ten (P<0.05). At week five, the wave crest on the axes Z, X, and Y, and the crest-trough difference on the axes X and Y were more prominent in the Gongfa group than in the manipulation group, showing statistical significance (P<0.05). At week ten, the wave crest, wave trough, and crest-trough differences on the axes Z, X, and Y were more prominent in the Gongfa group than in the manipulation group, presenting statistical significance (P<0.05). Conclusion: Practicing Shaolin Neigong can help the trainees reach the level of strength and frequency of Tuina clinicians in a shorter time in the forearm Rou-Kneading manipulation training. It can advance the efficiency in studying the forearm Rou-Kneading manipulation and promote the quality of the manipulation.
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Objective: To observe the clinical efficacy of tuina at head and abdomen for headache due to phlegm turbidity. Methods:A total of 56 patients with headache due to phlegm turbidity were randomized into a tuina group and a Chinese medicine group by the random number table, with 28 cases in each group. The tuina group was treated mainly with tuina at the head and abdomen, while the Chinese medicine group was treated with oral administration of Ban Xia Bai Zhu Tian Ma Tang(Pinellia, Atractylodes Macrocephala and GastrodiaDecoction). The course of treatment was 30 d. The scores of headache index, traditional Chinese medicine syndrome scale, and the therapeutic efficacy were observed. Results:There were 2 dropouts in each group during treatment. The total effective rate was 92.3％ in the tuina group, significantly higher than 76.9％ in the Chinese medicine group (P<0.05). The scores of headache index and traditional Chinese medicine syndrome scale in both groups decreased after treatment (bothP<0.05), and scores in the tuina group were lower than those in the Chinese medicine group (bothP<0.05). Conclusion:Tuina mainly at head and abdomen is effective in treating headache due to phlegm turbidity, and has a better effect than Ban Xia Bai Zhu Tian Ma Tang (Pinellia, Atractylodes Macrocephala and GastrodiaDecoction).
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Based on the advocacy coalition framework,the game between policy alliances with different beliefs is regarded as a key factor contributing to the long-term policy changes. At the same time, the policy changes are seen as the product of the antagonism of the core faith of the alliance, the policy-oriented learning and the internal and external shocks of the coalitions. The change of China's health care policy has always been accompanied by the debate on whether fairness or efficiency should be given priority between"government-led"and"market-oriented"coalitions. Its evolving characteristics are consistent with the explanatory logic of the advocacy coalition framework. Based on the advocacy coalition framework this paper makes a comprehensive of review of the historical literature on Chinese medical and health policy,it founds out that advocacy coalitions with different beliefs are critical to the re-form and return of the health care policy, policy-oriented learning promotes the gradual change of the policy, while internal and external shocks provide opportunities for policy paradigm change. Besides,the more passive and lack of active initiatives of policy change,the government's bias of the public welfare feature of the medical business coupled with the underdevelopment of market economy are the main causes for lagging medical policy and frequent industry chaos. To this end,it is recommended to highlight the forward-looking policy,strengthen the internal drive,create a more liberal debate environment for the coalitions,promote the learning of policy orientation among coalitions to en-hance the scientific and effective medical and health policy change.
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<p><b>OBJECTIVE</b>To summarize and critically assess the effificacy of Eastern and Western manipulative therapies for the treatment of neck pain in adults.</p><p><b>METHODS</b>A search of PubMed/MEDLINE, the Cochrane Central Register of Controlled Trials, ClinicalTrials.gov, EMBASE, etc. from their inception date to January 2014 with Chinese, Japanese, and Korean databases. Two reviewers independently selected randomized controlled trials (RCTs) with negative control or blank control, extracted data and assessed methodological quality. Meta-analysis and levels of evidence were performed by Revman5.1 and Grades of Recommendations Assessment, Development and Evaluation (GRADE) approach.</p><p><b>RESULTS</b>Nineteen clinical trials with adequate randomization were included in this review, 11 of them had a low risk of bias. The primary outcome for short-term pain had no significant differences, however, the secondary outcome, only the Numerical Pain Rating Scale (NPRS) score of intermediate-term [n=916, pooled mean differences (MD) =-0.29, P=0.02], the Neck Disability Index (NDI) score of short-term (n=1,145, pooled MD=-2.10, P<0.01), and intermediate-term (n=987, pooled MD=-1.45, P=0.01) were signifificantly reduced with moderate quality evidence. However, it supported the minimally clinically important difference (MCID) of the Visual Analogue Scale and NPRS pain score to be 13 mm, while NDI was 3.5 points. The meta-analysis only suggested a trend in favor of manipulative therapy rather than clinical signifificance.</p><p><b>CONCLUSIONS</b>The results do not support the existing evidences for the clinical value of Eastern or Western manipulative therapy for neck pain of short-term follow-up according to MCIDs. The limitations of our review related to blinding, allocation concealment and small sample size.</p>
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To assess the clinical effect and safety of Chinese traditional medicine injection combined with radiotherapy for esophageal cancer. The relative randomized controlled trials (RCTs) of Chinese medical injections (CMI) combined with radiotherapy as well as simple radiotherapy for esophageal cancer were searched from PubMed, Cochrane Library, EMBASE, Chinese Biomedical Literature Database(CBM), China National Knowledge Infrastructure (CNKI), Wanfang Database and VIP Database as at September 2014. Two researchers completed the data extraction and quality evaluation independently. The data were analyzed by GeMTC 0.14.3 and Stata 12. 0 software. Finally, 43 RCTs involving 3 289 patients were finally included. The star network was constructed by different comparison groups. The results of network meta-analysis showed that the seven CMIs combined with radiotherapy was superior to simple radiotherapy in the treatment of esophageal cancer in efficacy, quality of life, and reduction in the incidence of nausea and leucopenia, but with no significant difference among the seven CMIs. Probability ranking result showed a great possibility for Shenqi Fuzheng and astragalus polysaccharide injections in improving the overall response rate and quality of life, which were followed by cinobufagin and kangai injections. However, only one study was included for Shenqi Fuzheng and astragalus polysaccharide injections. Therefore, cinobufagin or kangai injections were preferred in improving the overall response rate and quality of life. Aidi or compound sophora injections were better than other CMIs in reducing? the incidences of nausea (III-IV) and leukopenia. More RCTs of Shenqi Fuzheng and astragalus polysaccharide injections combined with radiotherapy for patients with esophageal cancer were expected in the future to confirm our results. Moreover, study findings will be reported, particularly for the adverse events in radiotherapy for esophageal cancer.
Subject(s)
Adolescent , Adult , Aged , Aged, 80 and over , Child , Female , Humans , Male , Middle Aged , Young Adult , Combined Modality Therapy , Drugs, Chinese Herbal , Esophageal Neoplasms , Drug Therapy , RadiotherapyABSTRACT
<p><b>OBJECTIVE</b>To evaluate the antitumor efficacy of streptavidin-tagged interleukin-4 (IL-4-SA) bifunctional fusion protein in the immunotherapy of mouse model of superficial bladder cancer.</p><p><b>METHODS</b>IL-4-SA fusion protein was prepared and its biological activity was determined. One day after MB49 cell implantation, 100 µl of 1 mg/ml NHS-PEO4-biotin was instilled into the bladder for 30 minutes, followed by intravesical instillation of 100 µl PBS, GFP-SA+IL-4 or IL-4-SA and incubation for 1 hour. The bladder irrigation was performed twice a week for three weeks. The CTL cytotoxicity and profile of CD8(+) tumor-infiltrating lymphocytes were analyzed.</p><p><b>RESULTS</b>The IL-4-SA fusion protein was durably anchored to the biotinylated mucosal surface of bladder wall for up to 5 days.On day 80 after the implantation of MB49 cells, all of PBS-treated mice died, and 8 out of 10 mice in the GFP-SA-treated group died from tumor burden.In contrast, 5 out of 10 mice in the IL-4-SA-treated group were tumor-free. The MB49 tumor-specific cytotoxicity from mice in the IL-4-SA group was (11.3 ± 1.2)%, (22.7 ± 1.5)% and (31.0 ± 3.0)% at the effector to target ratios of 1:1, 25:1 and 50:1, respectively. But the corresponding cytotoxicity was (4.3 ± 0.6)%, (9.0 ± 1.0)% and (14.3 ± 1.5)% in the GFP-SA+IL-4 group, and (3.3 ± 0.6)%, (7.3 ± 0.6)%, (12.7 ± 2.1)% in the PBS group. The tumor-specific cytotoxicity in the SA-CD40L group was significantly higher than that in the control groups (P < 0.05). The infiltrating CD8(+) T cells in tumors in the IL-4-SA-treated group were increased compare with those in other groups.</p><p><b>CONCLUSION</b>Intravesical anchoring of IL-4-SA elicites strong and long-lasting immunoprotection against superficial bladder cancer, and the novel immunotherapy may be an attractive therapeutic alternative in future.</p>
Subject(s)
Animals , Humans , Male , Mice , Administration, Intravesical , Biotinylation , CD8-Positive T-Lymphocytes , Pathology , Cell Line, Tumor , Immunotherapy , Methods , Interleukin-4 , Metabolism , Therapeutic Uses , Mice, Inbred C57BL , Neoplasm Transplantation , Recombinant Fusion Proteins , Metabolism , Therapeutic Uses , Streptavidin , Metabolism , Therapeutic Uses , Urinary Bladder Neoplasms , Metabolism , TherapeuticsABSTRACT
<p><b>OBJECTIVE</b>To prepare reference samples of Mycobacterium tuberculosis culture filtrate protein-10 (CFP-10) and CFP10-streptavidin fusion proteins (CFP10/SA) for time-resolved fluoroimmunoassay (TRFIA).</p><p><b>METHODS</b>The CFP10 gene was amplified by PCR from Mycobacterium tuberculosis strain H37Rv and cloned into pET24b, pET24b-streptavidin (SA) or pET21a-SA expression vectors. The recombinant proteins CFP10, CFP10-SA and SA-CFP10 were expressed in Rosetta cells, purified via nickel affinity chromatography and refolded by dialysis. The sensitivity and stability of the resultant proteins as reference samples were evaluated by double-antibody sandwich TRFIA.</p><p><b>RESULTS</b>CFP10-SA and SA-CFP10 fusion proteins were expressed as inclusion bodies, whereas CFP10 was expressed in a soluble form. The resultant purity of the 3 recombinant proteins all exceeded 95%. TRFIA results showed that CFP-SA fusion protein possessed the best sensitivity (0.02 µg/L) and stability.</p><p><b>CONCLUSION</b>The reference samples of CFP10 for TRFIA detection have been successfully prepared and can be used in the development of a diagnostic kit for Mycobacterium tuberculosis.</p>
Subject(s)
Bacterial Proteins , Genetics , Reference Standards , Fluoroimmunoassay , Methods , Gene Amplification , Mycobacterium tuberculosis , Reference StandardsABSTRACT
<p><b>OBJECTIVE</b>To establish a method for organotypic slice culture of neonatal rat cortex in a modified condition and investigate the effect of spatial signals on neural stem cell (NSC) differentiation.</p><p><b>METHODS</b>The brain slices (200 µm in thickness) of neonatal SD rats (3 to 5 days old) were prepared and cultured in modified serum-free DMEM/F12 medium at 37 degrees celsius; with 95% O(2) and 5% CO(2). The organotypic slice cultures were observed regularly. NSCs isolated from the cortex of rat embryos (14-15 embryonic days) were cultured in serum-free DMEM/F12 supplemented with B27 and N2, and the passage 3 NSCs were labeled by CM-DiI before transplanted onto the organotypic slices cultured for 2 weeks. The survival of transplanted NSCs was assessed, and the cell differentiation was identified by immunofluorescence staining.</p><p><b>RESULTS</b>The organotypic slice cultures were well maintained for at least 4 weeks in the modified medium. The thickness of the organotypic slices reduced from 200 µm to 130 µm after 2-week culture in vitro due to the migration of the cells on the edge of the slices. CM-DiI-labeled NSCs survived well and differentiated into GFAP(+) glia and β-tubullin III(+) neurons.</p><p><b>CONCLUSION</b>Neonatal rat organotypic brain slice can be successfully cultured in a modified condition to serve as a model for studying NSC differentiation induced by spatial signals.</p>
Subject(s)
Animals , Rats , Animals, Newborn , Cell Differentiation , Physiology , Cerebral Cortex , Cell Biology , Coculture Techniques , Methods , Fetus , Neural Stem Cells , Cell Biology , Transplantation , Organ Culture Techniques , MethodsABSTRACT
<p><b>BACKGROUND AND OBJECTIVE</b>Intravesical administration of Bacillus Calmette-Guerin (BCG) after transurethral resection is by far the most effective local therapy for superficial bladder cancer, the fifth most common cancer in the world. However, approximately one-third of patients fail to respond and most patients eventually relapse. In addition, there are pronounced side effects of BCG therapy, such as BCG sepsis and a high frequency of BCG-induced cystitis. This study established a novel immunotherapy through immobilization of streptavidin-tagged human IL-2 (SA-hIL-2) on the biotinylated mucosal surface of bladder wall.</p><p><b>METHODS</b>A mouse orthotopic model of MB49 bladder cancer was established by perfusing MB49 cells into mouse bladders. The SA-hIL-2 fusion protein was immobilized on the biotinylated mucosal surface of the bladder wall. Treatment began on day 1 after MB49 implantation, once every 3 days for 6 times. Immunohistochemical assay was performed to assess the persistence of SA-hIL-2 immobilized on the biotinylated mucosal surface of the bladder wall. The mice were monitored for tumor growth and survival. On day 60 after MB49 implantation, the SA-hIL-2-cured mice, which were found to have no hematuria or palpable tumors, were challenged with wild-type MB49 cells implanted into the pretreated bladder and monitored for survival.</p><p><b>RESULTS</b>SA-hIL-2 could be immobilized efficiently and durably on the bladder mucosal surface as long as 7 days. On day 60 after MB49 implantation, 9 out of 20 SA-hIL-2-treated mice survived, but all mice in PBS control group died. More importantly, 5 out of 9 tumor-free mice in the SA-hIL-2 group were protected against a second intravesical wild-type MB49 tumor challenge.</p><p><b>CONCLUSIONS</b>SA-hIL-2 fusion protein could significantly inhibit tumor growth and extend the survival time in the orthotopic model of MB49 bladder cancer.</p>
Subject(s)
Animals , Female , Mice , Biotinylation , Cell Line, Tumor , Immobilized Proteins , Metabolism , Therapeutic Uses , Immunotherapy , Methods , Interleukin-2 , Metabolism , Therapeutic Uses , Mice, Inbred C57BL , Mucous Membrane , Metabolism , Neoplasm Transplantation , Receptors, Interleukin-2 , Metabolism , Recombinant Fusion Proteins , Metabolism , Therapeutic Uses , Streptavidin , Metabolism , Therapeutic Uses , Urinary Bladder , Pathology , Urinary Bladder Neoplasms , Allergy and Immunology , TherapeuticsABSTRACT
<p><b>OBJECTIVE</b>To explore the inhibitory effect of norcantharidin (NCTD) on the expression of DNA replication initiation protein Cdc6 in cancer cells.</p><p><b>METHODS</b>MTT assay was performed to detect the inhibitory effect on different cancer cell lines, including HeLa, HepG2, Jurkat and Ramos cells. The effect of NCTD on Cdc6 protein level was detected by Western blotting, and BrdU incorporation assay was used to evaluate the DNA replication of the cells.</p><p><b>RESULTS</b>NCTD significantly inhibited the proliferation of the cells and caused degradation of Cdc6 protein to result in the inhibition of the DNA replication of the cells shown by BrdU incorporation assay.</p><p><b>CONCLUSION</b>NCTD can induce the degradation of Cdc6 in cancer cells to produce an anti-cancer effect.</p>
Subject(s)
Humans , Bridged Bicyclo Compounds, Heterocyclic , Pharmacology , Cell Cycle Proteins , Metabolism , Cell Line, Tumor , DNA Replication , Nuclear Proteins , MetabolismABSTRACT
<p><b>OBJECTIVE</b>To obtain streptavidin-tagged human interleukin-21 (hIL21) fusion protein and evaluate its bioactivities.</p><p><b>METHODS</b>hIL21-SA-pET21 and pET24a-SA- hIL21 plasmids were constructed and expressed in BL21(DE3) host bacteria. The hIL21-SA and SA- hIL21 fusion protein were purified through Ni-NTA affinity chromatography and refolded by dialysis. Flow cytometry was used to detect hIL21-SA and SA- hIL21 fusion protein on the biotinylated MB49 tumor cells. MTT assay was used to evaluate the effect of the fusion protein on the proliferation of human peripheral blood lymphocytes (PBLs) stimulated by Anti-CD3.</p><p><b>RESULTS</b>The recombinant fusion proteins were highly expressed in BL21(DE3) at about 30% of the total bacterial proteins. The two fusion proteins exhibited bifunctional activities, i.e. both biotin-binding property and hIL21 activity and SA-mediated high-affinity binding to biotinylated cell surfaces (with anchoring modified rate of about 95.18% and 96.91%).</p><p><b>CONCLUSION</b>We have successfully obtained bifunctional fusion protein hIL21-SA and SA- hIL21,which will provide a basis for further study of tumor biotherapy using the proteins.</p>
Subject(s)
Humans , Cancer Vaccines , Allergy and Immunology , Cell Line, Tumor , Escherichia coli , Genetics , Metabolism , Interleukins , Genetics , Recombinant Fusion Proteins , Genetics , Allergy and Immunology , Streptavidin , GeneticsABSTRACT
<p><b>OBJECTIVE</b>To explore the effect of Chinese herbs during the perioperative period of laparoscopic cholecystectomy (LC).</p><p><b>METHODS</b>Three hundred and sixty patients of chronic lithic cholecystitis (LCCT) were randomly assigned to two groups by lottery, 180 patients in each group. During the peri-operative period, the control group was treated with conventional Western medicine and placebo. The treated group was given the same conventional Western medicine and Chinese herbal decoctions, with Shitong mixture No. 1 added before LC, and Liujunzi decoction added after LC for three days. The operation time, body temperature after LC, white blood cell count, wind-breaking time after operation, as well as the changes of tongue coating in the first three post-operative days were recorded.</p><p><b>RESULTS</b>There was no significant difference between the two groups in operation time (P>0.05), while the improvement in body temperature recovery, wind-breaking time and changes of tongue coating in the treated group were better than those in the control group (P<0.01).</p><p><b>CONCLUSION</b>Applying Chinese herbs during perioperative period of LC could effectively benefit early recovery in such patients.</p>