Objective To investigate the construction of Streptococcus mutans luxS gene knockout mutant which can act as the technical platform for following researches on luxS quorum sensing function in oral ecosystem.Methods Erythromycin resistance gene was inserted between two 1 kb fragments containing regions of DNA immediately upstream and downstream of the luxS translational start and stop codons.The resuhing construct was linearized and electro-transformed into Streptococcus mutans cells.After allelic exchange,the luxS gene knockout mutant strains were selected on 10μg/ml erythromycin plates,and compared the growth and biofilms formation of luxS knockout mutant with wild type strains.Results The luxSknockout mutant was confirmed by PCR,and it was also confirmed that this gene mutant could be stably passed through in vitro.The growth mode of luxS knockout mutant showed obvious difierences against that of wild type at stationary phase,the knockout mutant gained more bacteria cells growth.Conclusion Streptococcus mutans luxS gene has been successfully disrupted with allelic exchange.This mutant strains showed higher growth abilitv which could be the consequence of quorum sensing mutant.

Objective The discovery of microRNA (miRNA) in maternal serum has opened up new possibilities for non-invasive prenatal diagnosis.However,our understanding of these pregnancy-related miRNA in the serum of pregnant women with fetuses with neural tube defects (NTDs) is still limited.This article is to study the dysregulated expression of microRNA-423 (miR-423) in the serum of pregnant women with neural tube defect(NTD) fetuses and its potential role as a biomarker for non-invasive prenatal diagnosis of fetal NTD.Methods Thirty-three pregnant women whose fetuses were diagnosed as neural tube defects by ultrasound (22 cases of spina bifida and 11 cases of anencephaly)and 33 normal pregnant women were selected.Peripheral venous blood of each pregnant woman was obtained early in the morning,the serum was purified from blood by centrifugation,then total RNA was isolated from serum and the miR-423 levels were detected by real-time RT-PCR.The ROC curve was used for assessing the diagnostic accuracy of miR-423 for fetal NTD.Results We revealed miR-423 with signifcant down-regulation in expression in serum of pregnant women with NTD fetuses (0.96 ±0.14) compared as women with normal pregnancies(2.28 ±0.43) (P ＜0.05).We performed ROC analysis of data from the 33 case-control pairs.The expression of miR-423 could distinguish NTD cases from normal controls,with an AUC of 0.711 (95 ％ CI:0.566 ～ 0.856) (P ＜ 0.05).Moreover,the expression of miR423 decreased only in serum of pregnant women with anencephaly fetuses(0.58 ±0.08)by the analysis in different forms of NTD.Conclusion miR-423 is deregulated in the serum of pregnant women with NTD fetuses and highlight the clinical potential of miR-423 as biomarker for diagnosis and prognostication of fetal NTD.

Objective To investigate the effect and mechanism of phosphorylated protein kinase R-like ER kinase(p-PERK) and C/EBP homologous protein(CHOP) after hypoxic-ischemic brain damage ( HIBD) . Methods Neonatal 7-day-old Sprague Dawley rats were divided into sham-operation control group and HIBD group( n=30 per group) . Each group was divided into 0 h,6 h and 24 h subgroup after operation ( n=10 per group) . The ratio of apoptosis of brain cell was measured by flow cytometer and the expression of p-PERK and CHOP were detected by Western blot. Results (1)Apoptosis cell appeared at 6 h in HIBD group,the ratio of cell apoptosis was(2. 17 ± 0. 19)%. The apoptosis cell obvious increased at 24 h,the ratio of cell apoptosis was(13. 42 ± 0. 83)%. There was a significant increase in the ratio of apoptosis after HIBD 6 h and 24 h, as compared with sham-operation control group [ ( 0. 57 ± 0. 06 )%( P <0. 01 ) ] . ( 2 ) The expression of both p-PERK and CHOP was very low in sham-operation control group. In the HIBD group,the expression of both p-PERK and CHOP began to increase at 6 h and increased furthermore at HIBD 24 h. The differences in the expression levels of p-PERK and CHOP in HIBD group among different time points were significant( P<0. 01 ) . ( 3 ) The expression of p-PERK positively correlated with the expression of CHOP (r=0. 997,P< 0. 05). Conclusion With the emerging of apoptosis after HIBD,the expression of both p-PERK and CHOP increases. The imbalance in the expression of PERK induces the apoptosis of brain cells in the HIBD of neonatal rats by regulation of CHOP expression.

In either type 1 or type 2 diabetes,there is significant loss of β cell mass.Understanding the changing of β cell mass in the course of diabetes would provide important information for diagnosis and treatment.Functional imaging like magnetic resonance imaging or positron emission tomography (PET) can provide safe and noninvasive detection of loss of β cell mass in the course of diabetes.Among them,radio-labelled imaging is the most sensitive imaging procedure of the β cell ; For dihydrotetrabenazine PET imaging aiming β cell mass,there has been some primary outcome.For the key factor causing type 2 diabetes,18 F-6-deoxy-6-fluoro-D-glucose (18F-6-FDG) PET imaging is an effective tracer to study the glucose transporting condition in vivo.Further development of functional imaging will be of great value in diagnosis and treatment of diabetes.

Objective To investigate the serum levels change of vascular endothelial growth factor (VEGF) during operation of non-small-cell lung cancer (NSCLC). Methods 120 cases of NSCLC patient diagnosed by pathology as well as with operation indication were selected as the experimental group. The patients selected were required without any chemotherapy or radiotherapy before operation, besides that, they should have good compliance and free will to be examined. During the process of experiment, 60 cases concluded as healthy in the physical examination were chosen as control group. The correlative information of the experimental group were collected including periphery blood specimen collected in 3 days before the operation, and of the 1st day, 7th day and 30th day after the operation while the periphery blood specimen of control group were collected. The serum levels of VEGF were detected by adopting enzyme linked immunosorbent assay (ELISA) method. Results The serum levels of VEGF in NSCLC patients before operation, of the first postoperative day, of the seventh postoperative day and of the thirtieth postoperative day were significantly higher than that in healthy people (P＜0.01), respectively (279.14±44.89)μg/L, (282.70±42.74) μg/L, (353.79±44.55) μg/L, (178.40±43.43) μg/L and (91.40±16.55) μg/L. The serum levels of VEGF in NSCLC patients showed positive correlation with the stages (P＜0.01). Conclusion The serum levels of VEGF in NSCLC patients scale up. At the same time it shows positive correlation with the stages of the primary tumor. The serum levels of VEGF in NSCLC patients scales up by degrees in one week after the operation, and drop one month later.

OBJECTIVETo evaluate the in vitro cario-static effect of Galla chinesis with biofilm model.

METHODSA four-organism bacterial consortium was cultured in a biofilm model on hydroxyapatite (HA) discs in a continuous culture system and exposed to repeated solution pulsing. There were three groups with different solution pulsed in the model: negative control group was pulsed with distilled water, positive control group was pulsed with 100 mmol/L sucrose solution and experimental group was pulsed with 100 mmol/L sucrose solution containing 4.0 g/L Galla chinensis. During the experiment, the dynamic changes of pH were recorded. After 6 pulses, surface structure of the biofilm was observed with a scanning electron microscope and the population on the biofilm was enumerated.

RESULTSGalla chinesis significantly inhibited the adherence of Actinomyces naelundii to HA disc compared with the control group and facilitated the removal of acid products. It was also found that the extra-cellular polysaccharide was reduced with the pulsing of Galla chinesis.

CONCLUSIONGalla chinesis in the biofilm model can partially reduce the cario-genic response of sucrose solution.

Actinomyces ; drug effects ; physiology ; Bacterial Adhesion ; drug effects ; Biofilms ; growth & development ; Cariostatic Agents ; pharmacology ; Dental Caries ; microbiology ; Drugs, Chinese Herbal ; pharmacology ; Durapatite ; Humans ; Models, Biological ; Streptococcus mutans ; drug effects ; physiology ; Streptococcus sanguis ; drug effects ; physiology ; Sucrose ; pharmacology

OBJECTIVETo evaluate the Nidus vespae's cario-static effect on biofilm model in vitro.

METHODSA four-organism bacterial consortium was grown in a biofilm model on hydroxyapatite (HA) discs in a continuous culture system and exposed to repeated solution pulsing respectively. There were three parallel-connected flow cells in the model, so were the three groups with different solution pulsed in. Negative control group was pulsed with distilled water, positive control group was pulsed with 250 mmol/L sucrose solution as well. While 4.0 g/L Nidus vespae together with 250 mmol/L sucrose solution was pulsed in the experiment group. During the experiment, the pH responses against the pulses were recorded. After the 6 pulses, the biofilm surface structure was observed with a scan electron microscope and the population on the biofilm was enumerated.

RESULTSNidus vespae can significantly inhibit the adherence of Streptococcus mutans to HA discs compared with the control group of 250 mmol/L sucrose pulsed in, and can facilitate the remove of acid products. It is also found that the extra-cellular polysaccharide is reduced with the pulsing of Nidus vespae.

CONCLUSIONNidus vespae in the biofilm model can partially decrease the cariogenic response of sucrose solution pulsed in.

Anti-Infective Agents ; pharmacology ; Bacterial Adhesion ; drug effects ; Biofilms ; growth & development ; Dental Caries ; microbiology ; Dental Enamel ; metabolism ; Dental Plaque ; microbiology ; Durapatite ; Evaluation Studies as Topic ; Humans ; Models, Biological ; Propolis ; pharmacology ; Streptococcus mutans ; drug effects ; physiology ; Streptococcus sanguis ; drug effects ; physiology ; Sucrose ; pharmacology

Objective: To investigate the clinicopathologic features and genetic profile of large cell lung carcinoma (LCC) redefined by new classification. Methods: Basing on 2015 WHO classification criteria in redefining large cell lung carcinoma, the expression of specific markers (TTF1, Napsin A, p40, CK5/6, CK, vimentin and ZEB1) was detected by immunohistochemistry and D-PAS staining in 303 surgically-removed lung specimens previously diagnosed as large cell lung carcinoma. The clinicopathologic and genetic characteristics (including EGFR, KRAS, BRAF, ALK and ROS1 gene mutation) were analyzed. Results: Based on the new definition of LCC, 116 cases (116/303, 38.3%) of LCC formerly diagnosed were reclassified as solid adenocarcinoma, 49 cases (49/303, 16.2%) as squamous cell carcinoma, 6 cases (6/303, 2.0%) as adenosquamous carcinoma, 22 cases (22/303, 7.3%) as spindle cell carcinoma and only 110 cases (110/303, 36.3%) as large cell carcinoma. Redefined LCCs were characterized as middle-age (range 40-80), male (102/110, 92.7%) and smoking patients (64/110, 58.2%) with intermediate-advanced stage. Among 110 cases, 9 cases with EGFR mutation and 10 cases with KRAS mutation and 1 case with ALK fusion were found. No BRAF and ROS1 alterations were identified. Conclusions According to the new classification, LCCs formerly diagnosed are mostly reclassified as adenocarcinoma and non-keratinizing squamous cell carcinoma. The newly defined LCC may significantly benefit from clinical therapy.

Objective To investigate the effect of restrictive intravenous fluid on the complications and quality of life in the advanced hepatocellular carcinoma patients.Methods Clinical data of Three hundred and fifty-seven cases with advanced hepatocellular carcinoma from Mar 2010 to Mar 2016 was retrospectively analyzed.One hundred and sixty-eight cases were recruited in the restrictive intravenous fluid (RIF) group,and One hundred and eighty-nine cases were involved in the control group.The average volume of intravenous fluid of each day,plasma albumin concentration,splanchnocoel hydrops rate,phlebitis,incidence of vomiting,cancer related pain degree,anxiety degree were compared in the two groups.Results The average volume of intravenous fluid of each day in the RIF group [(720.29 ± 106.84) ml] were much lower than that in the control group [(1 820.36±342.12)ml] (P ＜0.05).The plasma albumin concentration in the RIF group [(35.65 ± 2.21)g/L] were higher than that in the control group [(32.25 ±2.32)g/L] (P ＜0.05).The rate of splanchnocoel hydrops,phlebitis,vomiting,bedsores,and hypstatic pneumonia in RIF group were 6.25％,4.69％,8.59％,3.9％,11.72％ and those in the control group were 13.97％,10.92％,17.47％,10.04％,and 24.45％,respectively (P ＜ 0.05).Moreover,the scores of cancer related pain and anxiety were much lower in the RIF group (5.21 ± 1.09,39.12 ± 5.54) than those in the control group(5.68 ± 1.18,41.56 ± 6.78) (P ＜ 0.01).Conclusions Restrictive intravenous fluid therapy can decrease the cancer associated complications and improved the quality of life in the advanced hepatocellular carcinoma patients.

Objective:To investigate the application and curative effect of fat grafting in supraorbital area in double blepharoplasty in patients with negative vector of the eyelid- globe.Methods:From January 2019 to January 2020, 78 patients who came to Fuzhou Meilai Huamei Beauty Hospital for double blepharoplasty were collected as patients with negative eyelid- globe vector. The patients were randomly divided into 2 groups, with 42 cases in the combined group and 36 cases in the simple group. The combined group underwent double blepharoplasty combined with fat grafting in the supraorbital area, and the simple group only underwent double blepharoplasty.Results:The postoperative follow-up was 3-12 months, with an average follow-up of 6 months. The efficacy of the combined group and the simple group was compared. There was no surgical complications in both groups. In improving the eyelid-globe vector, the combined group was higher than that of the simple group. The overall satisfaction of the combined group was higher than that of the simple group. Among them, the satisfaction of eye appearance, self-confidence, mental health, surgical results, decision-making and facial rejuvenation were all higher than those of the simple group, and there was no difference in the impact on early life between the two groups.Conclusions:Double blepharoplasty undergoing fat grafting in the supraorbital area at the same time are safe and conforms to the aesthetic standard. The patients are satisfied with the postoperative effects.