OBJECTIVETo mimic oral squamous cell carcinoma (OSCC) cell hypoxia by using chemical agent CoCl2 and to investigate its biological behaviour.

METHODSOral squamous cell carcinoma cell lines HSC-3 and SCC-4 were exposed to different concentration of CoCl2. HSC-3 and SCC-4 cells were treated with 50, 100, 150, 200 µmol/L CoCl2. Expression of hypoxia inducible factor-1α (HIF-1α), vascular endothelial growth factor (VEGF) and B-cell lymphoma-2 (BCL-2) were measured by real time polymerase chain reaction (PCR) and Western blotting in both mRNA and protein level. Cell proliferation, cell apoptosis and cell cycle were detected to analyze its biological behaviour. Both wound healing and Transwell assay were applied to test the ability of cell igration.

RESULTSThe result showed that after treatment of 150 µmol/L CoCl2 for 24 h, mRNA level of HIF-1α, VEGF and Bcl-2 was increased by 6.00 ± 0.20, 5.40 ± 0.40, 5.40 ± 0.30 (SCC-4); 5.60 ± 0.30, 5.20 ± 0.60, 5.80 ± 0.40(HSC-3). OSCC cells treated with 150 µmol/L CoCl2 for 24 h were collected. Compared with control group, the growth rate of cells was significantly decreased, P value was less than 0.05 (when HSC-3, SCC-4 cultured for 2 and 3 days). The apoptosis of OSCC cells was increased when treated with 150 µmol/L CoCl2 for 24 h:HSC-3 2.25% (control group) and 5.82% (treatment group); SCC-4 2.58% (control group) and 10.27% (treatment group). The migration ablility of OSCC cells was decreased when using 150 µmol/L CoCl2 for 24 h. The migration area ratio was (31.5 ± 2.3) % (HSC-3), (29.1 ± 1.5) % (SCC-4) in control group and (18.3 ± 1.9) % (HSC-3), (13.2 ± 0.8)% (SCC-4) in treatment group (P < 0.05).

CONCLUSIONSThe hypoxic cell model of OSCC could be induced by CoCl2. The expression level of hypoxic markers was up regulated significantly and the cells biological behaviour changed including decreased cell proliferation, increased apoptosis and decreased migration.

Apoptosis ; Blotting, Western ; Carcinoma, Squamous Cell ; metabolism ; pathology ; Cell Cycle ; Cell Hypoxia ; Cell Line, Tumor ; Cell Movement ; drug effects ; Cell Proliferation ; Cobalt ; Humans ; Hypoxia-Inducible Factor 1, alpha Subunit ; metabolism ; Mouth Neoplasms ; metabolism ; pathology ; Proto-Oncogene Proteins c-bcl-2 ; metabolism ; RNA, Messenger ; metabolism ; Up-Regulation ; Vascular Endothelial Growth Factor A ; metabolism

Objective To evaluate the ability of contrast-enhanced ultrasound (CEUS) as a prognostic indicator of renal function in chronic kidney disease (CKD) patients.Methods A total of 122 patients with CKD were collected,and patients with allergies to sulfur hexafluoride,pregnancy,cardiopulmonary insufficiency,urinary calculus and tumour were excluded.These patients were divided into estimated glomerular filtration rate [eGFR,ml·min1· (1.73 m2)-1] ≥60 group,eGFR 30-59 group and eGFR ＜ 30 group.CEUS was performed after an intravenous bolus injection of 1.5 ml SonoVue (BR1;Bracco Milan,Italy).Time-intensity curves (TICs) and quantitative indexes were created using QLAB quantification software.Followed up for 2 years,and patients with eGFR dropped 50％,double serum creatinine and end-stage renal disease (ESRD) were regarded as having kidney failure events.Risk factors related to kidney survival were investigated using a multivariate Cox regression model.Results One hundred patients were enrolled in the study,with 78％ patients in CKD 1-2 stages,16％ in CKD 3 stage and 6％ in CKD 4-5 stages.Patients were followed for a mean period of 14.1 months,ten (10％) patients exhibited composite kidney failure events.Among 3 groups,significant differences in the left kidney length derived peak intensity (DPI) were noted (P=0.014,P=0.010).Multivariate Cox regression analysis revealed that the DPI was an independent factor of progression of kidney disease.Multiple linear regression showed that age,basic eGFR,peak intensity were associated with eGFR decline rate.Patients with DPI ＜ 12.27 db were less to recover from kidney disease progression as compared with patients with DPI≥ 12.27 db (P=0.008).The area under the curve (AUC) for DPI was 0.778(95％ CI 0.612-0.944,P＜ 0.05),with a sensitivity of 64％ and a specificity of 88％.Conclusions The DPI might be the most valuable CEUS parameter for the evaluation of renal function.The DPI could serve as an independent predictor of the long-term prognosis of CKD patients.

Objective To investigate the current status of semen testing in the hospitals without assisted reproductive technology in Hu-nan Province,propose improvement strategies to enhance the diagnosis of male infertility and promote reproductive health services in these hospitals.Methods Questionnaire surveys and on-site investigations were conducted to examine the semen testing status in 67 hospitals without assisted reproductive technology in Hunan Province.The existing problems were summarized,and improvement strate-gies were suggested.Results Among the 67 hospitals,59.7％(40/67)performed semen testing.Of these,45％(18/40)possessed dedicated personnel,60％(24/40)possessed staff received relevant training,and 22.5％(9/40)held certificates.Only 30％(12/40)conducted sperm morphology testing,among them 20％(8/40)able to stain and interpret following WHO standards.The hospitals of 37.5％(15/40)used computer-assisted sperm analysis(CASA)systems,and 22.5％(9/40)were equipped with phase-contrast microscopes.30％(12/40)had dedicated examination areas,and 32.5％(13/40)had independent ejaculation rooms,17.5％(7/40)used disposable slides to observe sperm concentration.In 2022,the hospitals of 57.5％(23/40)had an average daily sample volume＜1.The hospital of 67.5％(27/40)performed standard operating procedures,40％(16/40)followed the WHO 5th la-boratory manual for routine testing,15％(6/40)conducted internal quality control,and 12.5％(5/40)participated in external quali-ty assessment(EQA).Another 55％(22/40)wished to participate in EQA.Conclusion The semen testing capacity in the hospitals,in which assisted reproductive technology is not yet carried out currently,urgently requiring multifaceted improvements.The proposed strategies include emphasizing semen testing,establishing reproductive medicine consortia for comprehensive support,establishing a provincial quality control center for EQA,founding a professional committee for a learning platform,offering training to enhance staff expertise,and including the pre-pregnancy semen testing in public health programs.

China has entered the task stage of comprehensive medical insurance payment reform, but there are problems restricting the development of innovative medical technology in the reform of diagnosis-related groups(DRG) payment system. The author introduced the international definition and scope of innovative medical technology, and summarized the preconditions and payment policy of short-term payment of innovative medical technology under the DRG payment system; And put forward suggestions in line with China′s actual situation, including clarifying the definition of innovative medical technology, setting access conditions for additional payment or actual payment, setting up special transition funds for high-value innovative drugs, clarifying the payment amount of innovative medical technology, and formulating payment strategies for innovative medical technology.

OBJECTIVETo investigate the associations of occupational, educational and residential factors with the outcomes of in vitro fertilization -embryo transfer (IVF-ET).

METHODSA retrospective analysis was conducted among 3998 patients undergoing the first treatment with IVF-ET and 3064 perinatal infants in our center between January, 2010 and June, 2014. The relation of the patients' occupation, educational background, and residence registry with IVF-ET outcomes was analyzed.

RESULTSThe risk of abortion in the first trimester was significantly higher in female patients engaged in agriculture (OR=4.319, P=0.047). Occupation and educational background of the patients and their spouse were related with the incidence of ectopic pregnancy following IVF-ET. The total incidence rate of perinatal birth defects was 2.1% in this cohort, and was the highest in Shenzhen (5.5%) but this difference was not statistically significant (P>0.05).

CONCLUSIONThe patients' occupation and educational background are related with the outcomes of IVF-ET, suggesting the importance of health education and prenatal examination.

Abortion, Spontaneous ; Educational Status ; Embryo Transfer ; Female ; Fertilization in Vitro ; Humans ; Incidence ; Infant ; Occupations ; Pregnancy ; Pregnancy Outcome ; Pregnancy, Ectopic ; Residence Characteristics ; Retrospective Studies

Objective To investigate the influence of different end shapes of esophageal stents on the mechanical behavior of the stent-esophagus system. Methods Through finite element simulation, the mechanical behavior of the coupling system between braided esophageal stents with different end shapes (straight-tube-shaped, cup-spherical-shaped, double-trumpet-shaped) and the esophageal cavity was analyzed. The influences of bare stents and covered stents with three different end shapes on stress distributions in the inner wall of the esophagus and on dilatation of the esophageal stenosis zone were compared. Results The reduction in stenosis rate caused by the bare stent was larger than that of the covered stent. Moreover, the equivalent stress and the contact stress caused by the bare stent were much larger than those of the covered stent. Different end shapes had a significant influence on the stress occurring in the healthy esophageal zone. Stress concentration occurred in the zones where the esophagus contacted the central part of the cup-shaped end and the edge of the double-trumpet-shaped end. The braided esophageal stents with three different end shapes all had good apposition.Conclusions Different end shapes of stents cause different stress states in the esophagus. A larger stress involves a higher probability of occurrence of esophageal tissue hyperplasia, but a smaller possibility of stent migration. Therefore, understanding the effect of the end shapes on stent performance can provide important theoretical references for optimization design of the braided stent and its clinical selection.

Objective: To evaluate the clinical effect of a digital whole-process surgical guide for immediate implantation in the molar area. Methods : Twenty-six patients with molar extraction plans were accepted for preoperative CBCT and model construction. Computer software was used to design the ideal three-dimensional position of the implant. The control group of 13 patients underwent immediate implantation with a free hand operation, whereas the experimental group of 13 patients underwent preparation and implant insertion under the guidance of a surgical guide. Bone grafting was performed, and a good initial stability was achieved. After 5 to 6 months, osseointegration was achieved, and the final restoration was delivered. After surgery, the accuracy of the three-dimensional position of the implants was measured, and at the 6 month return visit, the modified Plaque Index (mPLI), modified Sulcular Bleeding Index (mSBI) and probing depth (PD) were measured. Results : In the control group and experimental group, the vertical errors at the top of the implants were 1.246 ± 0.072 mm and 0.628 ± 0.046 mm (t = 26.078, P < 0.001), respectively, and the horizontal errors were 1.563 ± 0.086 mm and 0.546 ± 0.056 mm (t = 35.813, P < 0.001), respectively; and the vertical errors at the root of the implants were 1.352 ± 0.042 mm and 0.532 ± 0.030 mm (t = 57.021, P < 0.001), respectively, and the horizontal errors were 1.645 ± 0.076 mm and 0.625 ± 0.072 mm (t = 35.086, P < 0.001), respectively. For the experimental group, the mPLI value was 0.923 ± 0.760, the mSBI value was 0.846 ± 0.689, and the PD value was 3.460 ± 0.713 mm, which were significantly lower than those of the control group. For the control group, the mPLI value was 1.769 ± 0.927 (t = 2.546, P = 0.018), the mSBI value was 1.692 ± 0.947 (t = 22.605, P = 0.016) and the PD value was (4.579 ± 0.475) mm (t = 4.709, P < 0.001). Conclusion A digital surgical guide plate can increase the precision of immediate implantation and the peri-implant health in the molar area.

【Objective】 To observe the effects of early postnatal immune activation (EPIA) on social behaviors of male and female mice, and to explore the possible role of the functional state of astrocytes and microglia in this process. 【Methods】 Using lipopolysaccharide (LPS) induced EPIA mice as study subjects, mice were divided into the male-control, male-model, female-control, and female-model groups, each containing 10 mice (n=10). Behavioral tests were performed at 25 - 32 days of age, and the social behavior ability of mice was evaluated by open field test, three-chamber sociability test, and marble burying test. The expression levels of GFAP, IBA-1, TLR4, and NFκB p65 in the cortex and hippocampus were detected by Western blot (n=3). 【Results】 In behavioral tests, social index significantly decreased in LPS treatment group (F=14.907, P<0.05). The interaction effect between treatment and sex was significant in the residence time (F=5.260, P<0.05) and the number of buried marbles (F=7.788, P<0.05). LPS treatment decreased the retention time of the central region in male mice (F=4.261, P<0.05), and increased the number of buried marbles in males (F=20.645, P<0.05). Western blot results showed that LPS treatment increased the expression of GFAP protein in the hippocampus (F=50.443, P<0.05) and cortex (F=30.116, P<0.05), as well as the expression of IBA-1 protein (F=21.844. P<0.05) and TLR4 protein (F=6.215, P<0.05) in the cortex. The results of NFκB p65 showed a significant interaction between treatment and sex in the cortex (F=6.558, P<0.05), and LPS increased the expression of NFκB p65 protein in the cortex in female mice (F=16.317, P<0.05). 【Conclusions】 EPIA is sufficient to induce sex-specific autism spectrum disorder (ASD)-like behavior and enhance astroglial and microglial reactivity in mice. ASD-like behavior induced by EPIA may be related to the TLR4/NFκB signaling pathway in the cortex.