摘要
ObjectiveTo explore the impact of Gegen Qinliantang(GQT) on the fecal short-chain fatty acids(SCFAs) metabolism in antibiotic-associated diarrhea(AAD) through targeted metabolomics. MethodA total of 240 SD rats were randomly divided into six groups(n=40, half male and half female), including blank group, model group, bifidobiogen group(0.15 g·kg-1), and GQT high-, medium-, and low-dose groups(10.08, 5.04, 2.52 g·kg-1), except for the blank group, clindamycin(250 mg·kg-1) was given to all groups by gavage for modeling every day for 7 d. After successful modeling, each administered group was gavaged with the corresponding dose of the drug, and the blank and model groups were gavaged with an equal volume of normal saline solution, 1 time/d, for 14 d. At 0, 3, 7, 14 d after the drug intervention, eight rats were randomly selected from each group, respectively. Gas chromatography-time-of-flight mass spectrometry(GC-TOF-MS) was used to perform targeted metabolomic analysis of SCFAs in the feces of rats, and partial least squares-discriminant analysis(PLS-DA) was applied to compare the differences in metabolic profiles between groups at different treatment times, and to compare the changes in the contents of SCFAs in rat feces between groups. ResultPLS-DA results showed that the blank group could be clearly distinguishable from the model group, with GQT exhibiting a closer proximity to the blank group after 7 d of treatment. After further analyzing the composition of SCFAs, it was found that the proportion of acetic acid increased and the proportions of butyric acid, valeric acid, hexanoic acid and isovaleric acid decreased in the model group compared with the blank group. After the treatment with GQT, the proportions of butyric acid, isobutyric acid, valeric acid, and isovaleric acid increased, and the proportions of acetic acid, propionic acid and caproic acid decreased. Subsequent differential analysis revealed that GQT could significantly improve the content of butyric acid, and had a certain retrogressive effect on the contents of valeric acid and hexanoic acid. ConclusionThe medium dose group of GQT can improve the contents of SCFAs in AAD feces after 7 days of treatment, which may be related to the improvement of the composition ratio of SCFAs and the contents of butyric acid, valeric acid and caproic acid.
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Objective: To observe the clinical efficacy of herbal cake-partitioned moxibustion for ulcerative colitis (UC) and elucidate its mechanism by targeting the vitamin D receptor (VDR) signaling pathway. Methods: A total of 63 patients with UC were randomly divided into an observation group (30 cases, treated with herbal cake-partitioned moxibustion) and a control group (33 cases, treated with sham herbal cake-partitioned moxibustion). Moxibustion treatment was performed at Qihai (CV6) and bilateral Tianshu (ST25) and Shangjuxu (ST37), 3 times per week for 12 weeks. The total effective rate, visual analog scale (VAS) score for abdominal bloating and pain, and hospital anxiety and depression scale (HADS) score were compared between the two groups. Enzyme-linked immunosorbent assay was used to detect the concentrations of serum C-reactive protein (CRP), 25-hydroxyvitamin D [25(OH)D], and interleukin-12 (IL-12)/interleukin-23 (IL-23) p40. Immunohistochemistry was used to observe the expression levels of VDR and regenerating gene Ⅳ (Reg Ⅳ) proteins in colonic mucosa. The expression levels of VDR, cytochrome p45027B1 (CYP27B1), and Reg Ⅳ mRNAs were detected by real-time fluorescence quantitive polymerase chain reaction. Results: After treatment, the total effective rate in the observation group was 86.7%, which was significantly higher than 51.5% in the control group (P<0.05). After treatment, the VAS scores for abdominal bloating and pain in the observation group were significantly decreased (P<0.01), as well as the HADS-depression subscale (HADS-D) and HADS-anxiety subscale (HADS) scores (P<0.05), while only the VAS score for abdominal pain in the control group was reduced (P<0.05), and the improvements of the scores in the observation group were more significant than those in the control group (P<0.05). After treatment, the serum CRP concentrations in both groups and the IL-12/IL-23 p40 concentration in the observation group were significantly decreased (P<0.05), and the concentrations in the observation group were lower than those in the control group (P<0.05). The expression levels of VDR protein and mRNA in the colon in both groups were all increased (P<0.01), and the expression levels of Reg Ⅳ protein and mRNA and CYP27B1 mRNA were all decreased in the two groups (P<0.05 or P<0.01); the improvements in the observation group were more notable than those in the control group (P<0.05 or P<0.01). Conclusion: Herbal cake-partitioned moxibustion can effectively alleviate abdominal pain and diarrhea in patients with UC, improve depression and anxiety disorders, and regulate the expression of related proteins in the VDR signaling pathway. The mechanism may be related to inhibiting intestinal inflammation by reducing the release of the proinflammatory cytokine IL-12/IL-23 p40.
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ObjectiveTo investigate the mechanism of Gegen Qinliantang(GQT) on the intestinal flora of antibiotic-associated diarrhea(AAD) by 16S rRNA sequencing and network pharmacology. MethodSixty SD rats were randomly divided into six groups(n=10), including blank group, model group, GQT high-, medium- and low-dose groups(10.08, 5.04, 2.52 g·kg-1) as well as Lizhu Changle group(0.15 g·kg-1), except for the blank group, each group was given clindamycin(250 mg·kg-1) by gavage once a day for 7 consecutive days. After successful modeling, the blank group and the model group were given equal volumes of normal saline by gavage. The other groups were given corresponding doses of drugs by gavage for 14 days. Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform(TCMSP) was used to screen the active components and targets of GQT, GeneCards, Online Mendelian Inheritance in Man(OMIM) database, Pharmacogenetics and Pharmacogenomics Knowledge Base(PharmGKB), DrugBank and DisGeNET were used to search for AAD disease targets. The drug-disease common targets were obtained by R software. STRING was applied to analyze the target protein-protein interaction, and Kyoto Encyclopedia of Genes and Genomes(KEGG) pathway enrichment analysis was performed. Then hematoxylin-eosin(HE) staining was used to observe the pathological changes of the colon, and 16S rRNA sequencing of AAD colon content flora structure further verified the results of network pharmacology. ResultThrough network pharmacology, it was found that 238 active components were screened from GQT and acted on 276 component targets, among which quercetin, puerarin, wogonin and apigenin were the main core components of GQT, 1 097 AAD disease targets and 127 drug-disease intersection targets. The protein-protein interaction network mainly included core targets such as protein kinase B1(Akt1), interleukin(IL)-6 and IL-1β, which were mainly enriched in the IL-17 signaling pathway. It was verified through animal experiments that compared with the blank group, the colon structure of the model group was seriously abnormal, the intestinal epithelial columnar cells were damaged, the goblet cells were reduced, and a large number of inflammatory cells were infiltrated. Compared with the model group, the colon structure of the GQT high-dose group improved, but there were still abnormalities, the colon structure of GQT medium- and low- dose groups and Lizhu Changle group improved significantly and reached the normal level. GQT could improve the structural diversity of AAD intestinal flora. At the phylum level, the abundance of Firmicutes was increased and the abundance of Bacteroidetes was decreased. At the genus level, the abundance of Lactobacillus was increased, and the abundances of Prevotella and Bacteroides were decreased. Among them, Lactococcus could be used as a biomarker for AAD treatment with GQT, and the prediction of functional metabolism of intestinal flora revealed that GQT could promote acetate and lactate metabolic pathways in the intestine. ConclusionGQT may activate IL-17 signaling pathway by acting on the targets of Akt1 and IL-6 through key components such as quercetin and wogonin, and improve the abundance of Lactococcus in the intestinal tract as well as acetate and lactate metabolic pathways, so as to play a role in repairing the intestinal barrier for the treatment of AAD.
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Objective:To explore the mechanism of acupuncture and moxibustion for ulcerative colitis(UC)from the perspective of the P2X7 receptor(P2X7R)-nucleotide-binding oligomerization domain(NOD)-like receptor protein 3(NLRP3)inflammasome pathway. Methods:Sprague-Dawley rats were randomly assigned to a normal(N)group,a model(M)group,a herb-partitioned moxibustion(HM)group,and an electroacupuncture(EA)group.For modeling,the rats drank 4%dextran sulfate sodium for 7 d.Rats in the HM group and EA group received 7 consecutive days of HM or EA at bilateral Tianshu(ST25),respectively.The histopathological change in colon tissue was observed by hematoxylin-eosin(HE)staining;immunofluorescence staining was used to detect the protein expression of related molecules in the colon tissue,and enzyme-linked immunosorbent assay was used to detect the concentrations or contents of related molecules in the serum and colon tissue.Wild-type(WT)and P2X7R gene knockout(KO)mice were used to construct UC models,histopathological changes in the colon tissue were observed by HE staining,and the NLRP3 protein expression in the colon tissue was observed by immunohistochemistry. Results:Compared with the N group,the colon histopathological score in the M group was significantly increased,and the protein expression of P2X7R,NLRP3,apoptosis-associated speck-like protein containing CARD(ASC),Caspase-1,interleukin-1β(IL-1β),and interleukin-18(IL-18)in the colon tissue and the protein levels of IL-1β and IL-18 in the serum were significantly increased(P<0.05).Compared with the M group,the histopathological scores of the colon in the HM group and the EA group were significantly decreased,and the protein expression levels of P2X7R,NLRP3,ASC,Caspase-1,IL-1β,and IL-18 in the colon tissue and the protein level of IL-18 in the serum were significantly decreased(P<0.05).After UC modeling,the colonic mucosal epithelial damage and inflammatory cell infiltration in P2X7R KO mice were less than those in WT mice,and the NLRP3 protein expression in the colon was also decreased compared with that in WT mice(P<0.05). Conclusion:HM and EA at Tianshu(ST25)may inhibit the protein activities of P2X7R,NLRP3,ASC,and Caspase-1 in the colon tissue of rats with UC,thereby reducing the downstream molecules IL-1β and IL-18 in the P2X7R-NLRP3 inflammasome pathway to relieve UC inflammation.
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Objective:To observe the effects of acupuncture and moxibustion on phosphatase and PTEN-induced putative kinase 1(PINK1)/Parkin signaling pathway in the midbrain substantia nigra of Thy1-α synuclein(αSyn)transgenic model mice with Parkinson disease(PD). Methods:Twenty-four Thy1-αSyn transgenic mice were randomly divided into a model group,an acupuncture group,an acupuncture + moxibustion group,and a Western medicine group.Six wild-type mice in the same litter were used as the wild-type group.In the acupuncture group,Baihui(GV20)and Yanglingquan(GB34)were selected for acupuncture.In the acupuncture + moxibustion group,Guanyuan(CV4)was added on the basis of the acupuncture group.The Western medicine group was given rapamycin intraperitoneal injection at a dose of 10 mg/(kg·bw).The wild-type group and the model group were fixed without intervention.The overall rod performance(ORP)score of mice was observed in each group.The immunohistochemical method was used to detect the tyrosine hydroxylase(TH)positive neurons in the substantia nigra of mice in each group.The αSyn was detected by the immunofluorescence chemical method.The expression levels of αSyn,microtubule-associated protein 1 light chain 3(LC3)-Ⅱ/LC3-Ⅰ,autophagy protein sequestosome-1/protein 62(SQSTM-1/p62),PINK1,Parkin,and ubiquitin-specific protease 30(USP30)proteins were detected by Western blotting assay.The expression levels of LC3B,p62,PINK1,Parkin,and USP30 mRNAs were detected by fluorescence quantitative polymerase chain reaction. Results:Compared with the wild-type group,the ORP score,the p62,PINK1,and Parkin protein expression levels decreased significantly(P<0.01),the PINK1 mRNA expression level decreased(P<0.05),while the protein and mRNA expression levels of USP30 increased(P<0.05)in the model group.Compared with the model group,the ORP score in the acupuncture group and the acupuncture + moxibustion group increased(P<0.05);the expression level of LC3-Ⅱ/LC3-Ⅰ protein in the acupuncture + moxibustion group and the Western medicine group increased(P<0.05);the protein expression levels of p62,PINK1,and Parkin increased(P<0.05),while the USP30 protein expression level decreased significantly(P<0.01)in the acupuncture group,the acupuncture + moxibustion group,and the Western medicine group;the Parkin mRNA expression level in the acupuncture group and the acupuncture + moxibustion group increased(P<0.05);the USP30 mRNA expression level in the acupuncture + moxibustion group decreased(P<0.05). Conclusion:Acupuncture and moxibustion regulate the related molecule expression levels of PINK1/Parkin signaling pathway in the Thy1-αSyn transgenic PD model mice and promote the autophagy degradation of αSyn to exert the protective effect of dopaminergic neurons.
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As an important part of tumor microenvironment, neutrophils are poorly understood due to their spatiotemporal heterogeneity in tumorigenesis. Here we defined, at single-cell resolution, CD44-CXCR2- neutrophils as tumor-specific neutrophils (tsNeus) in both mouse and human gastric cancer (GC). We uncovered a Hippo regulon in neutrophils with unique YAP signature genes (e.g., ICAM1, CD14, EGR1) distinct from those identified in epithelial and/or cancer cells. Importantly, knockout of YAP/TAZ in neutrophils impaired their differentiation into CD54+ tsNeus and reduced their antitumor activity, leading to accelerated GC progression. Moreover, the relative amounts of CD54+ tsNeus were found to be negatively associated with GC progression and positively associated with patient survival. Interestingly, GC patients receiving neoadjuvant chemotherapy had increased numbers of CD54+ tsNeus. Furthermore, pharmacologically enhancing YAP activity selectively activated neutrophils to suppress refractory GC, with no significant inflammation-related side effects. Thus, our work characterized tumor-specific neutrophils in GC and revealed an essential role of YAP/TAZ-CD54 axis in tsNeus, opening a new possibility to develop neutrophil-based antitumor therapeutics.
Subject(s)
Humans , Animals , Mice , Adaptor Proteins, Signal Transducing/metabolism , Transcription Factors/metabolism , Stomach Neoplasms/pathology , Neutrophils/pathology , Signal Transduction/genetics , YAP-Signaling Proteins , Tumor Microenvironment , Hyaluronan Receptors/genetics摘要
Heart failure is the leading cause of death worldwide. Compound Danshen Dripping Pill (CDDP) or CDDP combined with simvastatin has been widely used to treat patients with myocardial infarction and other cardiovascular diseases in China. However, the effect of CDDP on hypercholesterolemia/atherosclerosis-induced heart failure is unknown. We constructed a new model of heart failure induced by hypercholesterolemia/atherosclerosis in apolipoprotein E (ApoE) and LDL receptor (LDLR) dual deficient (ApoE-/-LDLR-/-) mice and investigated the effect of CDDP or CDDP plus a low dose of simvastatin on the heart failure. CDDP or CDDP plus a low dose of simvastatin inhibited heart injury by multiple actions including anti-myocardial dysfunction and anti-fibrosis. Mechanistically, both Wnt and lysine-specific demethylase 4A (KDM4A) pathways were significantly activated in mice with heart injury. Conversely, CDDP or CDDP plus a low dose of simvastatin inhibited Wnt pathway by markedly up-regulating expression of Wnt inhibitors. While the anti-inflammation and anti-oxidative stress by CDDP were achieved by inhibiting KDM4A expression and activity. In addition, CDDP attenuated simvastatin-induced myolysis in skeletal muscle. Taken together, our study suggests that CDDP or CDDP plus a low dose of simvastatin can be an effective therapy to reduce hypercholesterolemia/atherosclerosis-induced heart failure.
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Objective: To observe the effect of electroacupuncture (EA) at different time points during the perioperative period on the recovery of gastrointestinal function after gastrointestinal malignant neoplasms surgery. Methods: Sixty-three patients who needed radical surgery for gastrointestinal tumors were randomized into a control group, treatment group 1 (postoperative EA group), and treatment group 2 (intraoperative and postoperative EA group). The control group received surgery and conventional Western medicine treatment, and treatment groups 1 and 2 received additional EA treatment at different time points. The initial flatus time after the surgery, visual analog scale (VAS) score at different time points after the surgery, the proportion of using patient-controlled analgesia (PCA) after the surgery, and the times of adding analgesics were observed in the three groups. Results: The initial flatus time after the surgery was earlier in treatment groups 1 and 2 than in the control group (P<0.05); the difference between treatment groups 1 and 2 was statistically insignificant (P>0.05). The VAS score was lower in treatment group 2 than in the control group at 6, 12, 24, and 72 h after the surgery (P<0.05); the VAS score was lower in treatment group 1 than in the control group only at 72 h after the surgery (P<0.05). There were no significant differences in the rate of using PCA among the three groups (P>0.05). Regarding the times of adding analgesics, it was less in treatment group 2 than in the control group at 12 h after the surgery (P<0.05). Conclusion: Either EA during and after the surgery or only after the surgery can hasten the initial flatus and boost the recovery of gastrointestinal function in patients after radical resection of gastrointestinal neoplasms. Successive EA during and after the surgery should be superior to postoperative EA regarding the analgesic effect after the surgery.
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Objective: By analyzing the clinical characteristics, treatment options, and prognosis of different stages of sarcomatoid hepatocellular carcinoma (SHC), this study aimed to improve clinicians' understanding of SHC. Methods: The clinical data of 24 patients with SHC between January 2015 and December 2019 from First Affiliated Hospital of Zhengzhou University were retrospectively analyzed. There were 16 men and 8 women with a median age of 55 years. Results: The clinical symptoms were mainly abdominal pain and fever; 66.7% of the patients had hepatitis virus infection and 79.2% had liver cirrhosis. Patients with stage III- carcinoma accounted for 87.5% of the study population. The mean largest tumor diameter was 6.27cm. Patients who were followed up were treated at our hospital, of which one patient underwent liver transplantation, eight underwent surgery, three received locoregional therapy only, and the rest received best support care, chemotherapy, or targeted or combined immunotherapy. Computed tomography or magnetic resonance imaging showed abnormal findings, and biopsy or postoperative pathology was consistent with SHC. The median follow-up period was 4.8(1-39.7) months, and the median overall and disease-free survival of postoperative patients was 4.7 and 2.3 months, respectively. The median overall survival (OS) was 4.8 months for patients with stage III- carcinoma. Conclusions: For patients with stage I SHC, surgical resection is the preferred treatment. Postoperative adjuvant therapy can benefit patients. For patients with stage III- SHC, the benefits of surgery are limited. Systemic chemotherapy, oral targeted drug treatment, or local therapy is feasible for reducing the tumor burden. However, the prognosis of patients with SHC is generally poor, regardless of stage or therapeutic methods. Chemotherapy combined with targeted drugs and immunotherapy may become a new therapeutic direction for advanced SHC.
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Objective:To investigate the protective effect of chrysin on the newborn rats with ischemic hypoxic brain damage and its related mechanism.Methods:The rats were randomly divided into sham-operated group, model group, low and high-dosage group of chrysin with 12 rats in each group. Except sham-operated group, the rats in other three groups were used as hypoxic-ischemic encephalopathy model. After the modeling, the rats in the low and high dosage groups were intraperitoneally injected with 30 mg/kg and 60 mg/kg chrysinimmediately, while the rats in the sham-operated group and the model group were intraperitoneally injected with equal volume of normal saline for 4 days. The HE staining was used to observe the protective effect of drugs on brain. The superoxide dismutase (SOD), catalase (CAT), malondialdehyde (MDA) levels in cells were measured by biochemical method. Western blot was used to detect the expression of heme oxygenase-1 (HO-1) and transcription factor NF-E2-related factor 2 (Nrf2).Results:The neurons in the cortex of rats in sham-operated group were arranged orderly with round nuclei and obvious nucleoli; the brain tissue of rats in model group showed edema and the arrangement of neurons was loose, the degeneration and necrosis of neurons in low and high dosage of chrysin groups were less than those in model group. Compared to the model group, the SOD activity (55.74 ± 5.14 U/mg, 69.84 ± 5.05 U/mg vs. 37.64 ± 6.41 U/mg) and CAT activity (2.44 ± 0.22 U/mg, 2.59 ± 0.42 U/mg vs. 2.08 ± 0.37 U/mg) in the brain tissue of rats in the low and high dose group were significantly increased ( P<0.05). The MDA level (3.74 ± 0.05 mmol/mg, 2.60 ± 0.18 mmol/mg vs. 4.35 ± 0.24 mmol/mg) in rat brain tissue of low and high dose group was significantly decreased ( P<0.05). The expressions of HO-1 (0.43 ± 0.08, 1.02 ± 0.15 vs. 0.14 ± 0.07) and Nrf2 (0.48 ± 0.07, 0.79 ± 0.09 vs. 0.26 ± 0.08) protein in rat brain tissue of low and high dose group were significantly decreased ( P<0.05). Conclusions:The chrysin could alleviate nerve injury in rats with hypoxic-ischemic encephalopathy, and its mechanism may be related to the up-regulated HO-1 and Nrf2 protein expression.
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As a pattern of lymph node metastasis , extracapsular lymph node involvement has occurred in many malignant tumors,including the esophageal carcinoma.This paper reviewed the research on extracapsular lymph node involvement in e-sophageal carcinoma worldwide including the development mechanism , clinical diagnosis, incidence and the prognosis signifi-cance, etc.The research progress and clinical significance of extracapsular lymph node involvement in esophageal carcinoma were fully explored and summarized.
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OBJECTIVE: To obtain Ginkgo biloba antimicrobial peptide (GBA) recombinant protein, and to investigate in vivo/in vitro antimicrobial activity of the protein so as to provide experimental basis for solving bacterial resistance and large-scale production of new plant-derived antimicrobial agents. METHODS: Based on gene technology, according to GBA gene sequence (FJ 865399) published by Genebank, recombinant expression vector plasmid pET32a(+)-GBA was constructed. Prokaryotic expression of recombinant protein was conducted by Escherichia coli, and then the protein was purified and identified by gel electrophoresis and Western blotting. Drug sensitivity of obtained recombinant protein to E. coli, Staphylococcus aureus, Pseudomonas aeruginosa, Salmonella typhimurium were investigated by Kirby-Bauer test. The minimum antibacterial concentration (MIC) and minimum bactericidal concentration (MBC) were determined by broth dilution method. The protective effects of recombinant protein on S. aureus infection model mice were investigated. RESULTS: Target recombinant protein was expressed successfully and purified (molecular weight of 32 kDa). The recombinant protein was moderately sensitive to S. aureus and low sensitive to other three bacterias. MIC and MBC of the recombinant protein to S. aureus were (50.00±5.00)mg/mL and(138.33±12.58)mg/mL, and MIC was significantly higher than those to other 3 kinds of bacterias (P<0.05). High-dose of recombinant protein (8.0 g/kg) could significantly reduce the S. aureus-induced mortality of mice (P<0.05), and had similar protective effect as positive drug penicillin. CONCLUSIONS: Obtained recombinant protein has obvious antimicrobial effects on S. aureus, inhibits E. coli and P. aeruginosa to certain extent and shows poor inhibitive effect on S. typhimurium. High-dose of recombinant protein shows significant protective effect for S. aureus infection model mice.
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There are so many biomechanical risk factors related with glaucoma and their relationship is much complex. This paper reviewed the state-of-the-art research works on glaucoma related mechanical effects. With regards to the development perspectives of studies on glaucoma biomechanics, a completely novel biomechanical evaluation factor -- Fractional Flow Reserve (FPR) for glaucoma was proposed, and developing clinical application oriented glaucoma risk assessment algorithm and application system by using the new techniques such as artificial intelligence and machine learning were suggested.
Subject(s)
Humans , Algorithms , Artificial Intelligence , Biomechanical Phenomena , Glaucoma , Diagnosis , Intraocular Pressure , Machine Learning , Risk Assessment , Risk Factors摘要
Objective To explore the application value of cone-shaped gastric tube combined with cervical end-to-end anastomosis in thoracoscopic and laparoscopic esophagectomy for esophageal cancer.Methods The retrospective and descriptive study was conducted.The clinical data of 122 patients with esophageal cancer who were admitted to West China Hospital of Sichuan University from December 2016 to December 2017 were collected.There were 89 males and 33 females,aged (61±8)years,with a range from 48 to 81 years.McKeowntype three-incision esophagectomy was performed,and the cone-shaped gastric tube was pulled up to esophagus in left neck for hand-sewn end-to-end anastomosis after the dissection of esophagus and stomach under total thoracoscopy and laparoscopy.Observation indicators:(1) surgical treatment situations;(2) postoperative complications;(3) follow-up.Follow-up using outpatient examination was performed to detect postoperative gastroesophageal reflux,anastomotic stenosis and evaluate anastomotic width at 1,3,6 months and one year postoperatively up to December 2018.Measurement data with normal distribution were represented by Mean±SD.Measurement data with skewed distribution were described by M (P25,P75) or M (range).Count data were expressed by absolute number.Results (1) Surgical treatment situations:122 patients underwent laparocopic McKeown-type three-incision esophagectomy successfully,using cone-shaped gastric tube combined with cervical hand-sewn end-to-end anastomosis as digestive tract reconstruction,with no intraoperative conversion to open surgery.The operation time,cervical anastomosis time,and volume of intraoperative blood loss were (229 ± 49) minutes,(27± 1) minutes,and 50 mL (40 mL,60 mL),respectively.There were 6-8 stations of lymph node dissected,and the number of lymph node dissected were 19 (15,25).Duration of postoperative hospital stay was 10 days (9 days,11 days) in the 122 patients.(2) Postoperative complications:31 of 122 patients had postoperative complications.The primary complications:3 patients with anastomotic fistula were cured by conservative treatment including enteral nutrition through placement of nutritional tube under gastroscope,closed thoracic drainage and anti-infection;6 cases with severe thoracic gastric dilation were cured after gastrointestinal decompression.The secondary complications of 22 patients included 8 cases with hoarseness caused by recurrent laryngeal never injury,5 with arrhythmia,9 with pulmonary infection.They were cured after symptomatic and supportive treatment.No chylothorax occured,and there was no perioperative death.(3) Follow-up:all the 122 patients were followed up for 10-24 months,with a median time of 19 months.During the follow-up,7 cases with anastomotic stenosis including 4 scoring less than grade 2 and 3 scoring more than grade 3 were relieved after dilation through gastroscope.There were 33 of 122 patients without any reflux symptoms,and 89 with reflux symptoms,among which 52 were scored 1,25 were scored 2 and 12 were scored 3.The width of gastroesophageal anastomosis measured by barium radiography at 1 month after operation was (1.2±0.4) cm.Conclusion Coneshaped gastric tube combined with cervical end-to-end anastomosis in digestive tract reconstruction of thoracoscopic and laparoscopic esophagectomy can reduce the incidence of postoperative anastomotic complications and thoracic gastric dilation,and nasogastric tube placement could be abandoned,which demonstrates good safety and universality.
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A set of device for the measurement of the pressure difference between the anterior and the posterior chambers (PDAP) was designed to investigate the temporal varying rules of PDAP in the anterior segment of rabbit eyes. A platform was established for the measurement of PDPA according to the mechanism of joint implement. Rabbit models with high intraocular pressure (IOP) were constructed by means of injecting Carbomer into anterior chamber to increase IOP. The 24 hours continuous measurements of PDAP were performed for normal rabbit eye and eye with high IOP. The developed device could sensitively response to the small pressure difference in eye. The pressure difference in the normal rabbit eye varied with time, and the variation range during a whole day was 5.84-96.84 Pa which reflected the existence of physiological rule. For the rabbit eye with high IOP, pressure in anterior chamber was higher than that in posterior chamber which was in consistence with the theory of self-adaptation adjustment. The present study indicates that the approaches and device designed in this paper can well implement the measurement of PDAP as well as the temporal varying rules of PDAP in the anterior segment during a whole day.
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Objective To construct anti-IL-4R murine anti-human single-chain variable fragment (scFvs) antibodies through BL21 (DE3) prokaryotic expression system. Methods The anti-IL-4R scFv sequence was optimizated on the basis of previous findings. The optimized scFv sequence was analyzed. The recombinant plasmid pET-32a-scFv was constructed. The recombinant plasmid was detected through enzyme identification, and was turned into BL21 (DE3) prokaryotic expression bacteria to express the pET-32a-scFv recombinant protein in E.coli BL21 (DE3). The purification and renaturation were researched, and SDS-PAGE analysis was studied. The molecular weight of ScFv against IL-4R was analyzed by SDS-PAGE. The expression of the fusion protein was detected by Western-blot assay. Results The length of fusion gene scFv-MLT sequence was 761 bp. The molecular weight of the recombinant expression of proteins of anti-IL-4R single antibody was approximately 45 ku. The recombinant proteins showed high specificity with anti-6 × His-tag antibody. Conclusion This experiment successfully constructs pET-32a-scFv prokaryotic expression system of recombinant protein with high immune reactivity, which provides the basis for further study of anti-IL-4R single chain antibody as drug target.
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OBJECTIVE:To adopt gel method for the determination of bacterial endotoxin in Fat emulsion(10%)/amino acid (15)/glucose (20%) injection. METHODS:According to the gel method in term ofbacterial endotoxin test methodin Chinese Pharmacopeia(2015 edition),the maximal valid dilution(MVD)of samples were determined through interference test and the vali-dated. The results were compared with chromogenic method. RESULTS:In gel method,the interference to agglutination reaction of TAL and bacterial endotoxin can be excluded when samples were diluted 24 times or less. In chromogenic method,the samples should be diluted 76 times or less. CONCLUSIONS:Gel method can be used for bacterial endotoxin test of Fat emulsion(10%)/amino acid(15)/glucose(20%)injection.
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BACKGROUND:Increasing evidence suggests that Sanguis draxonis is of great significance for treating pressure sores, burns and ulcers. OBJECTIVE:To observe the influences of Sanguis draxonis on vascular endothelial growth factor, epidermal growth factor, substance P and hydroxyproline in rats undergoing tissue-engineered skin transplantation and to verify its promotion of wound repair. METHODS:The tissue-engineered skin transplantation model of rats were established. Rats in treatment groups were given external application, single oral use of 0.1 g/(kg·d) Sanguis draxonis and combined use, respectively. No treatment was given in control group. After continuous treatment for 7 days, the expression levels of vascular endothelial growth factor, epidermal growth factor, substance P and hydroxyproline in skin tissue were determined. RESULTS AND CONCLUSION:Compared with the control group, the levels of vascular endothelial growth factor, epidermal growth factor, and hydroxyproline were significantly increased (P0.05). These results demonstrate that Sanguis draxonis can promote tissue-engineered skin to repair skin wound by upregulating the expression levels of vascular endothelial growth factor, epidermal growth factor, and hydroxyproline.
摘要
Objective:To explore the expressions of endogenous hydrogen sulfide (H2 S)and its synthases cystathionine beta synthase (CBS)and cystathionine gamma lyase (CSE)in the cell lines of normal bladder and bladder cancer,and to clarify their mechanism in the development of bladder cancer.Methods:The bladder cancer cell lines (5637,T24,UM-UC-3,EJ)and human bladder epithelial cell line SV-HUC-1 were selected.The expressions of CBS and CSE in bladder cancer and normal cell lines were analyzed by Western blotting assay and the productivities of H2 S in cell lines were detected by sensitive sulphur electrode assay.The EJ cells were selected based on the previous experimental results and divided into groups as follows:① 10 μmol· L-1 NaHS group, 50 μmol·L-1 NaHS group,100 μmol·L-1 NaHS group and control group.After drug treatment,the cell survival rate was measured by MTT assay at 24 and 48 h.② 5 μg·L-1 cisplatin group,cisplatin (5 μg·L-1 )+ NaHS (100 μmol·L-1 )group and control group.After medicine treatment,the cell survival rate was measured by MTT assay and the cell apoptotic rate was detected by flow cytometry at 48 h. Results:Compared with the normal bladder cells (SV-HUC-1),the expression levels of CBS and CSE and the productivity of H2 S in the bladder cancer cell lines (5637,T24,UM-UC-3 and EJ)were increased obviously (P <0.05 or P <0.01).Compared with control group,exogenous H2 S promoted the cell proliferation of EJ cells.The cell survival rates were increased with the increase of drug dose (P <0.05),which showed a dose-dependent effect.The cell survival rates were increased with the prolongation of time (P <0.05),which showed a time-dependent effect.After medicine treatment,compared with cisplatin group,the cell viability in cisplatin+NaHS group was increased (P <0.05)and the apoptotic rate was decreased (P <0.05).Conclusion:Endogenous H2 S and its synthases CBS and CSE have an increased expression level in bladder cancer cell lines compared with the normal bladder cells.H2 S can enhance the proliferation of bladder cancer cells and decrease the apoptosis induced by cisplatin.
摘要
OBJECTIVE:To establish the detection method for pyrogen-related factor interleukin 1β(IL-1β)through optimiz-ing detection condition,and to conduct preliminary methodology study. METHODS:The in vitro fresh human whole blood detec-tion method was used. The bacterial endotoxin standard solution(5,2,0.8,0.32 EU/ml)were added into diluted blood;using di-luted RPMI 1640 as blank control,the content of IL-1β in blood sample was determined by ELISA after incubation. The relation-ship of the addition of different attenuants(RPMI 1640 culture,sterilized normal saline)and fetal bovine serum,final dilution vol-ume fraction(40%,20%,10%and 8.3%)and storage duration(2,5,6,8,26 h)with the contents of endotoxin IL-1βwere in-vestigated,and related coefficient and detection limits were calculated. Different dilution times of Qingkailing injection and Ginaton injection samples and interference solutions were added into diluted blood to detect their recovery. RESULTS:The results indicated that RPMI 1640 media and 40% diluted blood was more sensitive(detection limit was 0.128 EU/ml,r=0.993);while the addition of fetal bovine serum didn’t influence the results. The detection limits of blood sample storied at 4 ℃ for 26 h were 0.128 EU/ml (r>0.990). When Qingkailing injection and Ginaton injection were diluted 10,32 and more times,the detection method was not interfered and the recovery ranged 68%-118%. CONCLUSIONS:Established in vitro fresh human whole blood detection method can be used for the detection of pyrogen,and provides trial evidence for the pyrogen detection of TCM injection.