OBJECTIVETo explore the differentially expressed genes and related signaling pathways in MC3T3-E1 osteo- blasts in response to suitable fluid shear stress values and action time with cDNA microarrays.

METHODSMC3T3-E1 cells cultured on a cover slip were subjected to fluid shear stress using a parallel plate flow chamber. The harvested RNA was used for microarray hybridization comprising approximately 44 170 genes, as well as for the subsequent real-time quantitative polymerase chain reaction validation of expression levels for selected genes. Microarray results were analyzed by using both GO and Pathway analysis.

RESULTSMicroarray analysis indicated that 884 differentially expressed genes were found. Among these genes, 444 were upregulated, whereas 440 were downregulated. The Notch signal and RIG- I -like receptor signaling pathways were involved in the Pathway analysis. GO analysis mainly involved different functional classifications, such as prostaglandin biosynthesis, nitric oxide-mediated signal transduction, calcium mediated signal, and cellular immune response, among others.

CONCLUSIONThe mechanism underlying the protective effect of fluid shear stress on MC3T3-E1 cells might be related to promoting cell survival- and inhibiting cell apoptosis-related signaling pathways and biological processes.

Apoptosis ; Calcium ; Humans ; Oligonucleotide Array Sequence Analysis ; Osteoblasts ; Signal Transduction ; Stress, Mechanical

OBJECTIVETo clarify the related factors of marginal bone loss (MBL) around tissue level implants in the posterior part of the mandible.

METHODSA total of 116 tissue level implants were implanted in the mandibular posterior region of 76 patients. Patients' information, including general characteristics, implant characteristics, implant site characteristics, and prosthesis characteristics, was recorded. Their cone beam computed tomography data were measured immediately after implant placement, 3 months later, and 3 and 12 months after prosthesis loading. The measurement of MBL was conducted by One Volume Viewer software. SPSS 20.0 was used for statistic analysis.

RESULTSSmoking, cortical bone thickness (CBT), collum angle (CA), and implant local sanitation showed significant differences with body mass loss (P<0.05). No significant differences were found among sex, age, length of implant, diameter of implants, implant systems, bone height, prosthesis type, and MBL (P>0.05).

CONCLUSIONThe risk factors that caused MBL were smoking, thicker CBT, larger CA, and poor implant local sanitation. Among them, poor implant local sanitation had the highest correlation with MBL.

Alveolar Bone Loss ; epidemiology ; etiology ; Cone-Beam Computed Tomography ; Dental Implants ; adverse effects ; Dental Prosthesis Design ; Dental Prosthesis, Implant-Supported ; Follow-Up Studies ; Humans ; Mandible ; Mandibular Prosthesis ; statistics & numerical data ; Oral Hygiene ; Postoperative Complications ; Smoking ; adverse effects ; Treatment Outcome

Objective To investigate the effects of physical and chemical factors in the environment for dried blood sample (DBS) preparation of neonatal screening assay.Methods A total of 60 normal and 120 positive DBS were prepared under control and 10 different conditions.Another 30 normal and 80 positive DBS were prepared under control and 7 different concentration gradients of formaldehyde.The levels of phenylalanine (Phe),glucose-6-phosphate dehydrogenease (G6PD),thyroid stimulating hormone (TSH) and 17α-hydoxyprogesterone (17α-OHP) were tested by time-resolved fluorescence immunoassay or fluorescence assay.Statistical analysis was performed using SPSS 22.0 software.Results Compared with the control group,the results of Phe were not significantly different (P ＞ 0.05) when the samples were dried under the formaldehyde sensitive threshold (4.62 to 6.95 ppm for 18 hours).G6PD levels were significantly lowered when the samples were dried under all the conditions except for fast cold drying (2 to 8 ℃ overnight and formaldehyde condition,0.30 to 0.38 ppm for 4 hours or 0.21 to 0.24 ppm for 18 hours).TSH and 17α-OHP levels were lowered obviously when the samples were dried under the conditions of humidity,UV and formaldehyde condition (TSH:0.32 to 0.52 ppm for 4 hours,0.38 to 0.45 ppm for 18 hours,17α-OHP:4.37 to 4.62 ppm for 4 hours,0.38 to 0.45 ppm for 18 hours).The results of Phe,G6PD,TSH and 17α-OHP were not statistically different with the control group when the samples were dried under the fast cold drying and 2 to 8 ℃ overnight.Conclusion The physical and chemical factors in the environment of DBS preparation should be related to the accuracy of neonatal disease screening closely.The necessary control factors including formaldehyde,ethanol,glacial acetic acid,ultraviolet irradiation,heat,humidity and decoration pollution may exhibit significant effects on the preparation of DBS.Fast cold drying and overnight at 2 to 8 ℃ could be available for DBS preparation.

Objective To establish a high purity primary culture methods of human nasal epithelial cells (HNEC) in vitro and to provide a successful primary culture model for evaluation experiments of the nasal preparation .Methods Primary culture of human nasal epithelial cells were performed with enzymatic dissociation of isolated tissue and cultured in serum-free medium .HNEC were separated through magnetic field by immunomagnetic beads .We determined the purity of the separated cells by light microscopy and flow cytometry .The morphology of HNEC was observed with a scanning electron microscope .Results Under an inverted phase microscope ,the cells morphology was paving stone shaped .Under the scanning electron microscopy ,abundant microvilli and cilia differentiation were observed .Flow cytometry showed the epithelial cells accounted for 99% .Conclusion The highly purified HNEC can be directly isolated by the magnetic cell sorting system .The cell model can be used for the basic research of nasal cavity preparation .

This study aimed at investigating the antiviral constituents from the active fractions of Tong-An (TA) injection.In this study,the active constituents of TA injection were screened by LPS-induced PGE2 production mode to detect the contents of PGE2.The chemical constituents were isolated by HP-20 macroporous resin,silica gel column chromatography,ODS column chromatography,Sephadex LH-20 column chromatography and preparative and semi-preparative HPLC.The structures were identified by spectral data and physicochemical property.As a result,the 95％ ethanol eluate of TA injection on the macroporous adsorption resin column was proved to be the active fraction of TA injection.Seventeen compounds were isolated from TA injection and identified as syringaresinol (1),N-Trans-Feruloyltyramine (2),chelerythrine (3),sinomenine (4),coptisine (5),sanguinarine (6),chelidoniny (7),magnoflorine (8),allocryptopine (9),protopine (10),farrerol (11),dihydrosanguinarine (12),heptadec-(9Z)-enoic acid (13),chlorogenic acid (14),cryptochlorogenin acid (15),3,5-di-O-caffeoylquinic acid (16) and 4,5-di-O-caffeoylquinic acid (17).PGE2 inhibitory activities of these compounds were determined,among which six compounds presented inhibitory activities against PGE2.It was concluded that all the isolated compounds from TA injection were firstly reported with the favorable inhibitory activities of compounds 2,5,9,10,11,12 against PGE2.

Objective:To investigate the therapeutic effect of Wenxin granule combined with sotalol on patients with arrhythmia and its influence on the levels of plasma angiotensin Ⅱ(Ang Ⅱ), C-reactive protein (CRP) and NT-proBNP.Methods:From January 2018 to October 2019, 162 patients with arrhythmia admitted to Taizhou Central Hospital were divided into observation group (81 cases) and control group (81 cases) according to the random digital table method.The control group was treated with sotalol, and the observation group was treated with Wenxin granule on the basis of the control group.The course of treatment in both two groups was 2 months.The changes of cardiac function, Ang Ⅱ, CRP and NT-proBNP levels, as well as adverse reactions were compared before and after treatment.Results:The total effective rate of the observation group (92.59%, 75/81) was higher than that of the control group (77.78%, 63/81) (χ 2=7.044, P<0.05). After treatment, the LVEDD [(53.18±3.09)mm] and LVESD [(52.19±2.35)mm] in the observation group were lower than those in the control group [(58.97±2.73)mm and (57.79±2.65)mm], while the SV [(76.93±3.67)mL] and LVEF [(53.90±2.64)%] in the observation group were higher than those in the control group [(71.35±2.87)mL and (46.75±2.18)%], the differences were statistically significant ( t=12.638, 14.230, 10.779, 18.795, all P<0.05). The plasma AngⅡ [(96.52±12.16)ng/L], CRP [(4.30±0.69)mg/L] and NT-proBNP [(394.25±47.15)ng/L] in the observation group were lower than those in the control group [(135.42±23.15)ng/L, (6.27±0.78)mg/L and (565.29±56.72)ng/L] ( t=13.389, 17.025, 20.870, all P<0.05). The incidence of adverse reactions in the observation group (11.11%) was lower than that in the control group (30.86%) (χ 2=9.529, P<0.05). Conclusion:Wenxin granule combined with sotalol is effective in the treatment of arrhythmia, which can reduce the plasma levels of Ang Ⅱ, CRP and NT-proBNP, improve the heart function and has no obvious adverse reactions.

Objective:To investigate the pathogenesis, precaution and treatment of neonatal congenital complete heart block (CCHB) in twins.Methods:The clinical data of a case of premature twins with neonatal CCHB from the Department of Neonatology, the First Affiliated Hospital of Xinxiang Medical University were retrospectively analyzed and related literature was reviewed.Results:(1)Case review: the 37-year-old gravida had no symptoms.Fetal ultrasound cardiogram(fUCG)at 23 weeks of gestation indicated bradycardia and CCHB.Then, the mother was diagnosed with undifferentiated connective tissue disease.After treatment with human immunoglobulin, dexamethasone and hydroxychloroquine, fUCG at 31 weeks of gestation still suggested CCHB.An emergency cesarean section was performed on the diagnosis of threatened preterm labor.With weakly positive neonatal antinuclear antibody (ANA), and positive Ro60 and Ro52 autoantibodies, twins were diagnosed with CCHB by 24 hour-Holter monitors.One of the twins was discharged with CCHB (ventricular rate of 80-90 times/min) after systemic therapy, but the weight increased to 2 200 g. The other one of the twins suffered from the sudden decrease of heart rate and blood pressure and finally died of sudden cardiac arrest.(2) Literature search: two cases in Chinese and 9 cases in English were reviewed.Among them, 9 cases were sjogren syndrome type A (SSA)/Ro and sjogren syndrome type B(SSB)/La related CCHB, and 2 cases were idiopathic CCHB.Conclusions:The placental transfer of anti-SSA or anti-SSB is an important mechanism of neonatal CCHB in twins, and other factors may also be involved.Current treatments are unsatisfactory.Most patients need pacemaker implantation.Early diagnosis and prenatal management can improve the prognosis.

Objective To investigate the feasibility of Region 4 Stork(R4S)project used for newborn screening by tandem mass spectrometry in China.Methods This retrospective study was performed among 362 822 neonates screened by tandem mass spectrometry from May 2015 to April 2016 in Zhejiang newborn screening center.Infants were grouped by screening result category: 83 true positive cases,360 554 true negative cases and 2 185 false positive cases.Raw data was uploaded into R4S website to perform postanalytical interpretive tools, then results were analyzed with interpretation rules.The comparisons of normal population percentiles were done at five selected percentiles between Zhejiang newborn screening center and R4S project with min-max normalization.Results Compared with cutoff system by using R4S project with interpretation rules,the positive predictive value increased from 3.7%to 8.3%,the specificity increased from 99.40%to 99.75%, and the false positive rate declined from 0.6% to 0.2%. The two cases of true positive hyperprolinemia were reported negative, and one case of β-ketothiolase deficiency was misdiagnosis.Totally 311 638 cases in true negative group were resolved by postanalytical interpretive tools,and the remaining 48 916 cases were excluded with interpretation rules.False positive cases were reduced to 897 cases.Results of percentiles comparison showed that levels of some markers were significantly different between zhejiang newborn screening center and R 4S project.Conclusions R4S project effectively improved the newborn screening performance, whereas leaded to a small number of misdiagnosis and missed diagnosis.Besides,many true negative cases should be excluded with interpretation rules.Optimization should be achieved based on local normal population.(Chin J Lab Med,2018,41:300-304)

Objective:To investigate the application feasibility of Region 4 Stork (R4S) system, an international collaborative newborn screening data platform, combined with cut-off value analysis in the neonatal screening for very long chain acyl-CoA dehydrogenase deficiency (VLCADD) by tandem mass spectrometry (MS/MS).Methods:The retrospective study was performed in 2, 040 072 neonates screened by MS/MS in Neonatal Screening Center of Zhejiang Province, China from October 2013 to July 2018. Nine hundred and ten cases were determined and identified as suspected positive VLCADD neonates by traditional cut-off method of tandem mass spectrometry. The original data of these 910 screened neonates were further analyzed by R4S system. Based on clinical diagnosis and ACADL gene test results, the screening efficiency between two methods was statistically compared.Results:The data of 910 suspected VLCADD-positive cases interpreted by cut-off method were further analyzed by R4S system, and the positive interpretation was reduced to 238 cases (including 9 confirmed positive cases). A total of 16 different mutations were found in ACADL gene sequencing among the confirmed children. The screening false positive rate (FPR) declined from 0.44‰ (901/2 040 072) to 0.11‰ (229/2 040 072), the rate of positive predictive value (PPV) increased from 0.99% (9/910) to 3.78% (9/238), and the specificity increased from 99.96% (2 039 162/2 040 063) to 99.99% (2 039 834/2 040 063). There was a statistically significant difference between cut-off method alone and cut-off method combined R4S system analysis (χ2=393.5, P<0.05). Conclusions:The R4S system combined with cut-off method applied in VLCADD neonatal screening by MS/MS can effectively improve screening performance, reduce false positive rate, and has certain value in clinical application.