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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 feasibility of ultrasound screening and diagnosis of fetal cleft palate in early pregnancy, analyze and summarize the imaging technology and image characteristics of two-dimensional and three-dimensional ultrasound in normal fetus and cleft palate fetus.Methods:A total of 10 519 pregnant women participated in the early pregnancy were included from January 2016 to June 2020 in Shenzhen Hospital, University of Chinese Academy of Sciences. The palatal line on the standard section of fetal nuchal translucency (NT) measurement was used as a screening marker for routine observation. For fetuses with abnormal palatine line, posterior nasal triangle of coronal plane and axial plane of maxillary alveolar arch of two-dimensional ultrasound were added as the diagnostic sections, and three-dimensional volume data of fetal face were collected, and three dimensional multimodal imaging technology was used to analyze the volume data off-line to determine or exclude fetal severe cleft palate. All fetuses were followed up during the second trimester for deformity scanning and post natal (or induced labor) assessment.Results:Of the 10 519 fetuses, the standard NT plane was obtained and the palatal line was observed in 10 204 cases(97.01%), with normal palatal line in 10 169 cases.In 35 suspected cases, 13 cases were confirmed cleft lip and palate by two and three dimensions ultrasound, and were confirmed by induced labor. There were 7 cases in unilateral side, 3 cases in bilateral, 2 cases in median cleft lip and palate, 1 cases in irregular cleft lip and palate, and no false positive results were reported. Twenty-two suspicious cases were excluded by increasing the two-dimensional sectional and three-dimensional volumetric off-line analysis, and screening after the second trimester and after birth. There was 1 case of missed diagnosis of simple cleft palate.Conclusions:Palatal line is a good screening marker for fetal cleft palate in early pregnancy. For fetuses with abnormal palatine line, the adding of posterior nasal triangle and the axial plane of maxillary alveolar arch, and combining three-dimensional volume data for off-line analysis can determine or exclude severe cleft palate. This study is of great significance for early screening and diagnosis of severe fetal cleft palate, prenatal genetic counseling and prevention birth defect.
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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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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 summarize the characteristics and associated malformation of fetal isolate cleft palate in prenatal ultrasonography, and analyze the reason of ultrasound misdiagnosis and missed diagnosis in isolate fetal cleft palate prenatally. Methods Systemic screening was performed with two-and three-dimensional ultrasonography in 3 576 cases. The fetal lip and plane were observed especially in nasolabial coronary plane, axial plane through maxilla, median sagittal plane, oblique coronal plane through oral cleft. Meanwhile the accompanied deformity were also screened. And prenatal ultrasound results were compared with postpartum ifndings. Results Eleven in 3 598 cases (0.31%, 11/3 598) were diagnosed as fetal isolate cleft palate by prenatal ultrasonography. The ultrasonic characteristics of isolate cleft palate were:(1) One case ofⅠ° cleft palate, the ultrasonic manifestations:in median sagittal plane, the hyperecho line of median palatine suture was disappeared, and the mucous membranes above and below it were complete;in oblique coronal plane of soft palate through oral cleft, the soft palate was complete and continuous;uvula couldn′t be displayed. (2) Three cases ofⅡ° cleft palate, the ultrasonic manifestations:in median sagittal plane though jaw, the hyperecho line of median palatine suture was shorter;the latter half and the midline of soft palate was disappeared;in both paramedian sagittal plane, the arc-shaped hyperecho line of median palatine suture were displayed;and longer than the hyperecho of midline of palate;in oblique coronal plane of hard palate through oral cleft, the ifrst half hyperecho line of hard palate was continuous, the middle of the latter half hyperecho line was interrupted;in oblique coronal plane of soft palate through oral cleft, the midline of soft palate was interrupted. 3D volume data analysis showed that the ifrst half hard palate was complete, the midline of the latter half hard palate and soft palate was interrupted. (3) Seven cases ofⅢ° cleft palate, the ultrasonic manifestations:in median sagittal plane, the hyperecho line of median palatine suture was disappeared;in oblique coronal plane of hard palate through oral cleft, the middle part echoes of the hard palate was interrupted;in oblique coronal plane of soft palate through oral cleft, the midline of soft palate was interrupted;oral and nasal cavity were communicated;the hyperecho of the vomer at the lower edge of the nasal septum could be displayed though oral cavity. 3D volume data analysis showed that hard palate and soft palate were interrupted. The hyperecho of the vomer at the lower edge of the nasal septum could be displayed clearly though oral cavity. Prenatal ultrasonic diagnosis was conifrmed by postpartum ifndings. And 2 cases were misdiagnosed (0.06%, 2/3 598), 1 case was missed diagnosed (8.33%, 1/12). The incidence of isolate fetal cleft palate was 0.33%(12/3 598). In 12 cases of isolate fetal cleft palate, 11 cases were accompanied with other fetal deformities, including central nervous system malformations (6/12), small jaw (6/12), urinary tract malformation (5/12), hydramnios (2/12), and absence of amniotic lfuid (1/12). Conclusions Fetal secondary palate should be routinely included in the prenatal screening. When secondary palate planes weresuccessfully demonstrated, the isolate cleft palate could be detected. Prenatal diagnosis of the isolate cleft palate is contributive to prenatal counseling and risk assessment.
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This study was aimed to prepare the spraying agent of prescriptions of Miao nationality herb and investigate the effect of Miao nationality herbs spray for serum SOD, MDA, and expression of Fas and Caspase-3 mRNA in lung tissues of silica-treated rats. The healthy SD rats were divided into 5 groups. Silica dust suspension was used in the model establishment of 4 groups. After the model was successfully established, 3 groups were randomly selected and given glucocorticoids atomization inhalation, Miao nationality herbs spray, Miao nationality herbs spray combined with intragastric administration of herbal medicine, respectively. After 40-day treatment, water-solubletetrazolium salt (WST-1) was used in the detection of serum superoxide dismutase (SOD). Thiobarbituric acid (TBA) was used in the detection of malondialdehyde (MDA). The mRNA expression variance of the Fas and Caspase-3 were detected by RT-PCR. The results showed that compared with the silica dust suspension group, the SOD activity of serum in the Miao nationality herbs spray group was significantly increased (P< 0.05). MDA content and the mRNA of Fas and Caspase-3 were significantly lower in the Miao nationality herbs spray group (P< 0.05). It was concluded that Miao nationality herbs spray group was able to increase the SOD activity of serum, decrease MDA content, and obviously decrease the expression of Fas and Caspase-3 of lung tissues among silica dust suspension rats.
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Objective To evaluate the value of angled insonation technique in obtaining the volumes of the fetal palate and in three-dimensional post-processing.Methods Five hundred and sixty-six cases of 20~28 weeks fetuses with normal face were confirmed by systemic ultrasound examinations and post-natal examination.The volumes in the standard midsagittal plane and angled midsagittal plane were obtained,then post-processed these data by 4D-View software.The same technology was used for volume data acquisition and processing to 1 case of cleft lip and palate fetal.Results In 566 fetuses,two groups of volume data were successfully acquired in 458 cases (standard midsagittal plane group) and 411 cases (angled midsagittal plane group).Fetal primary palate in the standard group and the angled group can be both well visualized(100%).Axial plane of hard palate can not be reconstructed in the standard group.Only 36% of the fetal hard palate's coronal plane and transoral oblique coronal plane can be reconstructed.The visualization rate of soft palate was 87% in the standard group,which was slightly higher than the angled group (82%).The visualization rate of hard palate's axial plane,coronal plane and transoral oblique coronal plane was 100% respectively in the angled group.Cleft alveolar process involving the secondary palate can be clearly visualized in 1 case of cleft lip and palate fetus.Conclusions By using the angled insonation technique,which is based on clearly visualizing secondary palate to obtain volumes,using three-dimensional post-processing technique can obtain the conventional three orthogonal planes and any other section of the whole palate.By using it we can assess the integrity of fetal palate,diagnose cleft lip with or without cleft palate and classify the cleft palate,even make prenatal diagnosis of the isolated cleft palate possible.
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Objective To explore the ultrasonic diagnosis of fetal cleft palate.Methods By applying coronal,sagittal and transverse planes to scan the fetal facial anatomy routinely, 2125 pregnant women with 20 odd weeks′ gestation were examined.Results Complete cleft palate was diagnosed in 4 cases by ultrasonography, all of which were accompanied with complete cleft lip and alveolar process.Incomplete cleft palate were missed in 2 cases.Conclusions Complete cleft palate is accompanied mostly with cleft alveolar process and complete cleft lip, and cleft palate can be found out through cleft alveolar process.To those fetuses without cleft lip or with incomplete cleft lip,their palate cannot be explored by ultrasound,and their cleft palate cannot be displayed.
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Forty-three patients who had ultrasonic examinations and operative resuIts selected from105 cases of painless obstructive jaundice were analysed.The results showed that 29 patients(67.4%>had diseases of head of pancreas,biliary duct cancer and carcinoma around ampuila and others. 5 pacalculus of common bile duct had no obvious biliary colic.The mecharlism of painless calculus was investigated. It is suggested that the exploratory laparotomy should be done as early as possible when there is difficulty to diagnose the cause of painless obstrutive jaundice by imaging technique.