[Feasibility study of expectant management of different degrees of vaginal fluid in pregnant women with premature rupture of membranes in the second trimester].

Objective: To investigate the feasibility of expectant management of different degrees of vaginal fluid in pregnant women with premature rupture of membranes in the second trimester. Methods: A retrospective cohort study was conducted to collect 103 pregnant women who were diagnosed with premature rupture of membranes in the second trimester of pregnancy and insisted on continuing the pregnancy in Shanxi Bethune Hospital from July 2012 to July 2022. According to the degree of vaginal fluid, pregnant women were divided into rupture group (with typical vaginal fluid, 48 cases) and leakage group (without typical vaginal fluid, 55 cases). The rupture latency (the time from rupture of membranes to termination of pregnancy), gestational weeks of termination, indications and methods of termination of pregnancy, maternal infection related indicators and perinatal outcomes were compared between the two groups. Univariate regression model was used to analyze the correlation between different degrees of vaginal fluid in pregnant women with premature rupture of membranes and maternal and neonatal outcomes. Results: (1) Obstetric indicators: there was no significant difference in the gestational age of rupture of membranes between the two groups (P>0.05). However, the proportion of rupture latency >28 days in the leakage group was significantly higher than that in the rupture group [42% (23/55) vs 13% (6/48); χ2=33.673, P<0.001], and the incidence of pregnancy termination ≥28 weeks was significantly higher [47% (26/55) vs 19% (9/48); χ2=9.295, P=0.002]. (2) Indications and methods of termination: the incidence of progressive reduction of amniotic fluid as the indication for termination in the leakage group was significantly lower than that in the rupture group [22% (12/55) vs 42% (20/48); χ2=4.715, P=0.030], and the incidence of full-term termination in the leakage group was significantly higher than that in the rupture group [31% (17/55) vs 12% (6/48); χ2=5.008, P=0.025], while there were no significant differences in the indications of termination of pregnancy, including amniotic cavity infection, uterine contraction failure and fetal distress between the two groups (all P>0.05). The incidence of induced labor or spontaneous contraction in the leakage group was significantly lower than that in the rupture group [53% (29/55) vs 81% (39/48); χ2=9.295, P=0.002], while the cesarean section rate and vaginal delivery rate were similar between the two groups (both P>0.05). (3) Infection related indicators: the incidence of amniotic cavity infection in the leakage group was significantly higher than that in the rupture group [31% (17/55) vs 13% (6/48); χ2=4.003, P=0.045]. However, there were no significant differences in the elevation of inflammatory indicators, the positive rate of cervical secretion bacterial culture and the incidence of tissue chorioamnionitis between the two groups (all P>0.05). (4) Perinatal outcomes: the live birth rate in the leakage group was significantly higher than that in the rupture group [51% (28/55) vs 27% (13/48); χ2=5.119, P=0.024]. The proportion of live births with 1-minute Apgar score >7 in the leakage group was significantly higher than that in the rupture group [38% (21/55) vs 17% (8/48); χ2=4.850, P=0.028]. However, there were no significant differences in the birth weight of live births and the incidence of neonatal complications between the two groups (all P>0.05). (5) Univariate regression analysis showed that compared with the rupture group, the leakage group had a higher risk of pregnancy termination at ≥28 gestational weeks (RR=2.521, 95%CI: 1.314-4.838; P=0.002), amniotic infection (RR=2.473, 95%CI: 1.061-5.764; P=0.025), perinatal survival (RR=1.880, 95%CI: 1.104-3.199; P=0.014). Conclusion: Compared with pregnant women with typical vaginal fluid in the second trimester of premature rupture of membranes, expectant treatment for pregnant women with atypical vaginal fluid is more feasible, which could effectively prolong the gestational weeks and improve the perinatal live birth rate.

Intermediate tidal volume is an acceptable option for ventilated patients with acute respiratory distress syndrome.

OBJECTIVE: Evidence only proves low surpasses high tidal volume (VT) for acute respiratory distress syndrome (ARDS). Intermediate VT is a common setting for ARDS patients and has been demonstrated as effective as low VT in non-ARDS patients. The effectiveness of intermediate VT in ARDS has not been studied and is the objective of this study. DESIGN: A retrospective cohort study. SETTING: Five ICUs with their totally 130 beds in Taiwan. PATIENTS OR PARTICIPANTS: ARDS patients under invasive ventilation. INTERVENTIONS: No. MAIN VARIABLES OF INTEREST: 28-D mortality. RESULT: Totally 382 patients, with 6958 ventilator settings eligible for lung protection, were classified into low (mean VT=6.7ml/kg), intermediate (mean VT=8.9ml/kg) and high (mean VT=11.2ml/kg) VT groups. With similar baseline ARDS and ICU severities, intermediate and low VT groups did not differ in 28-D mortality (47% vs. 63%, P=0.06) or other outcomes such as 90-D mortality, ventilator-free days, ventilator-dependence rate. Multivariate analysis revealed high VT was independently associated with 28-D and 90-D mortality, but intermediate VT was not significantly associated with 28-D mortality (HR 1.34, CI 0.92-1.97, P=0.13) or 90-D mortality. When the intermediate and low VT groups were matched in propensity scores (n=66 for each group), their outcomes were also not significantly different. CONCLUSION: Intermediate VT, with its outcomes similar to small VT, is an acceptable option for ventilated ARDS patients. This conclusion needs verification through clinical trials.

Intermediate tidal volume is an acceptable option for ventilated patients with acute respiratory distress syndrome / El volumen tidal intermedio es una opción aceptable para pacientes ventilados con síndrome de dificultad respiratoria aguda

Objective Evidence only proves low surpasses high tidal volume (VT) for acute respiratory distress syndrome (ARDS). Intermediate VT is a common setting for ARDS patients and has been demonstrated as effective as low VT in non-ARDS patients. The effectiveness of intermediate VT in ARDS has not been studied and is the objective of this study. Design A retrospective cohort study. Setting Five ICUs with their totally 130 beds in Taiwan. Patients or participants ARDS patients under invasive ventilation. Interventions No. Main variables of interest 28-D mortality. Result Totally 382 patients, with 6958 ventilator settings eligible for lung protection, were classified into low (mean VT=6.7ml/kg), intermediate (mean VT=8.9ml/kg) and high (mean VT=11.2ml/kg) VT groups. With similar baseline ARDS and ICU severities, intermediate and low VT groups did not differ in 28-D mortality (47% vs. 63%, P=0.06) or other outcomes such as 90-D mortality, ventilator-free days, ventilator-dependence rate. Multivariate analysis revealed high VT was independently associated with 28-D and 90-D mortality, but intermediate VT was not significantly associated with 28-D mortality (HR 1.34, CI 0.92–1.97, P=0.13) or 90-D mortality. When the intermediate and low VT groups were matched in propensity scores (n=66 for each group), their outcomes were also not significantly different. Conclusio Intermediate VT, with its outcomes similar to small VT, is an acceptable option for ventilated ARDS patients. This conclusion needs verification through clinical trials (AU)

Objetivo La evidencia solo demuestra que el volumen tidal (VT) bajo supera al alto para el síndrome de dificultad respiratoria aguda (ARDS). La VT intermedia es un escenario común para los pacientes con ARDS y se ha demostrado que es tan eficaz como la VT baja en pacientes sin ARDS. No se ha estudiado la eficacia de la VT intermedia en el ARDS y es el objetivo de este estudio. Diseño Un estudio de cohorte retrospectivo. Ámbito Cinco UCI con un total de 130 camas en Taiwán. Pacientes o participantes Pacientes con ARDS bajo ventilación invasiva. Intervenciones No. Variables de interés principales Mortalidad 28-D. Resultado Un total de 382 pacientes, con 6958 configuraciones de ventilador elegibles para protección pulmonar, se clasificaron en bajo (VT medio=6,7ml/kg), intermedio (VT medio=8,9ml/kg) y alto (VT medio=11,2ml/kg). Grupos de VT. Con un ARDS inicial similar y una gravedad en la UCI, los grupos de VT intermedia y baja no difirieron en la mortalidad 28-D (47% vs. 63%, p=0,06) u otros resultados como mortalidad 90-D, días sin ventilador, dependencia del ventilador índice. El análisis multivariado reveló que la VT alta se asoció de forma independiente con la mortalidad 28-D y 90-D, pero la VT intermedia no se asoció significativamente con la mortalidad 28-D (HR 1,34, IC 0,92-1,97, p=0,13) o la mortalidad 90-D. Cuando los grupos de VT intermedia y baja se emparejaron en puntajes de propensión (n=66 para cada grupo), sus resultados tampoco fueron significativamente diferentes. Conclusión La VT intermedia, con resultados similares a la VT pequeña, es una opción aceptable para pacientes con ARDS ventilados. Esta conclusión necesita verificación a través de ensayos clínicos (AU)

Intermediate tidal volume is an acceptable option for ventilated patients with acute respiratory distress syndrome.

Objective: Evidence only proves low surpasses high tidal volume (V T) for acute respiratory distress syndrome (ARDS). Intermediate V T is a common setting for ARDS patients and has been demonstrated as effective as low V T in non-ARDS patients. The effectiveness of intermediate V T in ARDS has not been studied and is the objective of this study. Design: A retrospective cohort study. Setting: Five ICUs with their totally 130 beds in Taiwan. Patients or participants: ARDS patients under invasive ventilation. Interventions: No. Main variables of interest: 28-D mortality. Result: Totally 382 patients, with 6958 ventilator settings eligible for lung protection, were classified into low (mean V T = 6.7 ml/kg), intermediate (mean V T = 8.9 ml/kg) and high (mean V T = 11.2 ml/kg) V T groups. With similar baseline ARDS and ICU severities, intermediate and low V T groups did not differ in 28-D mortality (47% vs. 63%, P = 0.06) or other outcomes such as 90-D mortality, ventilator-free days, ventilator-dependence rate. Multivariate analysis revealed high V T was independently associated with 28-D and 90-D mortality, but intermediate V T was not significantly associated with 28-D mortality (HR 1.34, CI 0.92-1.97, P = 0.13) or 90-D mortality. When the intermediate and low V T groups were matched in propensity scores (n = 66 for each group), their outcomes were also not significantly different. Conclusion: Intermediate V T, with its outcomes similar to small V T, is an acceptable option for ventilated ARDS patients. This conclusion needs verification through clinical trials.

Objetivo: La evidencia solo demuestra que el volumen tidal (VT) bajo supera al alto para el síndrome de dificultad respiratoria aguda (ARDS). La VT intermedia es un escenario común para los pacientes con ARDS y se ha demostrado que es tan eficaz como la VT baja en pacientes sin ARDS. No se ha estudiado la eficacia de la VT intermedia en el ARDS y es el objetivo de este estudio. Diseño: Un estudio de cohorte retrospectivo. Ámbito: Cinco UCI con un total de 130 camas en Taiwán. Pacientes o participantes: Pacientes con ARDS bajo ventilación invasiva. Intervenciones: No. Variables de interés principales: Mortalidad 28-D. Resultado: Un total de 382 pacientes, con 6958 configuraciones de ventilador elegibles para protección pulmonar, se clasificaron en bajo (VT medio = 6,7 ml/kg), intermedio (VT medio = 8,9 ml/kg) y alto (VT medio = 11,2 ml/kg). Grupos de VT. Con un ARDS inicial similar y una gravedad en la UCI, los grupos de VT intermedia y baja no difirieron en la mortalidad 28-D (47% vs. 63%, p = 0,06) u otros resultados como mortalidad 90-D, días sin ventilador, dependencia del ventilador índice. El análisis multivariado reveló que la VT alta se asoció de forma independiente con la mortalidad 28-D y 90-D, pero la VT intermedia no se asoció significativamente con la mortalidad 28-D (HR 1,34, IC 0,92-1,97, p = 0,13) o la mortalidad 90-D. Cuando los grupos de VT intermedia y baja se emparejaron en puntajes de propensión (n = 66 para cada grupo), sus resultados tampoco fueron significativamente diferentes. Conclusión: La VT intermedia, con resultados similares a la VT pequeña, es una opción aceptable para pacientes con ARDS ventilados. Esta conclusión necesita verificación a través de ensayos clínicos.

[Research and development of an air sampling tube for chlorobenzene compounds and its supporting determination method].

Objective: To establish a GDH-3 air sample tube for simultaneous determination of twelve kinds of chlorobenzene compounds (CBs) in workplace air by gas chromatography. And to established a matching determination method. Methods: In October 2020, the vapor and aerosol CBs in workplace air were collected by GDH-3 air sampling tube, and desorption and elution with 3.00 ml toluene for 15 min, then the solution separated by DB-23 capillary column, and finally detected with microcell electron capture detector. Results: The quantitative determination ranges of twelve isomers of CBs were 0.71×10(-3)-2000.00 mg/L, with the correlative coefficients were 0.99967-0.99998. The minimum detectable concentrations were 0.04-112.63 µg/m(3), and the minimum quantification concentrations were 0.14-375.42 µg/m(3) (15.00 L sample, 3.00 ml sample solution) . The average elution efficiencies were 96.00%-104.00%. The within-run relative standard deviations (RSDs) were 2.54%-6.12%, and the between-run RSDs were 3.85%-7.87%. Sealed samples could be stable at room temperature for at least 15 days. Conclusion: GDH-3 air sample tube can be used for simultaneous determination of twelve kinds of CBs in workplace air by gas chromatography. The established supporting measurement method meets the measurement requirements of the occupational health standard detection method, and the it's suitable for the simultaneous determination of 12 kinds of CBS in the air.

[Percutaneous biliary stent combined with brachytherapy for malignant obstructive jaundice: a multicenter retrospective controlled study].

Objective: To investigate the efficacy, safety and prognostic factors of percutaneous biliary stent combined with iodine-125 seed chain brachytherapy (radiotherapy) in the treatment of malignant obstructive jaundice. Methods: Data of 107 cases with malignant obstructive jaundice treated with percutaneous biliary stent implantation from January 2017 to December 2020 were retrospectively analyzed. Among them, 58 cases received biliary stent combined with iodne-125 seed chain brachytherapy (study group), and 49 cases received biliary stent implantation (control group). The changes of bilirubin, stent patency time, complications, overall survival (OS) and prognostic factors were analyzed in both groups. Results: The incidence of complications in the study group and the control group were 17.2% and 18.3% respectively, and the difference was not statistically significant (P=0.974). Serum total bilirubin levels were decreased significantly in both groups at one month after surgery (P<0.001). Postoperative stent patency time was significantly better in the study group (10.0±1.6 months) (95% CI: 8.2～12.5) than that in the control group (5.2±0.4 months) (95% CI: 4.1～6.0, P<0.001). The median OS was longer in the study group (11.2±1.8 months) (95% CI: 9.2～12.8) than that in the control group (8.0±1.1 months) (95% CI: 8.0～12.8, P<0.001). Multivariate analysis result showed that stent combined with brachytherapy (HR=0.08, 95% CI:0.04～0.15, P<0.001) and receiving further anti-tumor therapy after surgery (HR=0.27, 95% CI:0.15~0.49, P<0.001) were independent risk factors affecting the patency of biliary stents. Preoperative percutaneous transhepatic biliary drainage (HR=0.46, 95%CI:0.28～0.74, P=0.002), stent combined with brachytherapy (HR=0.23, 95%CI:0.14～0.39, P<0.001) and receiving further anti-tumor therapy after surgery (HR=0.37, 95%CI:0.22～0.61, P<0.001) were independent risk factors affecting OS. Conclusion: Percutaneous biliary stent combined with brachytherapy is safe and effective in the treatment of malignant obstructive jaundice, which can significantly prolong the patency time of biliary stent and the survival time of patients.

Dendritic cell TLR4 induces Th1-type immune response against Cryptosporidium parvum infection.

The objective of this study was to investigate the mechanism of Toll-like receptor (TLR4)- mediated dendritic cell (DC) immune against Cryptosporidium parvum infection. C. parvum sporozoites were labeled with 5,6-carboxyfluorescein diacetate succinimidyl ester. Murine bone marrow-derived DCs were isolated, and divided into TLR4 antibody blocking (TAB; infected with 2 × 105 labeled sporozoites and 0.5 µg TLR4 blocking antibody), TLR4 antibody unblocking (TAU; infected with 2 × 105 labeled sporozoites), and blank control (BC; with 1.5 mL Roswell Park Memorial Institute 1640 medium) groups. The adhesion of Cryptosporidium sporozoites to DCs and CD11c+ levels were examined by fluorescence microscopy and flow cytometry. Male KM mice were orally injected with C. parvum. The proliferation of T lymphocytes in spleen, expression of cytokines in peripheral blood, and TLR4 distribution features in different organs were further determined by immunohistochemistry. A significantly higher expression of CD11c+ and higher C. parvum sporozoite adhesion were found in the TAU group compared with other groups. The expression of CD4+CD8- /CD8+CD4- in the spleen were obviously differences between the TAB and TAU groups. The expression of TLR4, interleukin IL-4, IL-12, IL-18 and IFN-γ improved in the TAU group compared with TAB group. Higher expression of TLR4 was detected in the lymph nodes of mice in the TAU group, with pathological changes in the small intestine. Hence, TLR4 could mediate DCs to recognize C. parvum, inducing Th1 immune reaction to control C. parvum infection.

Dendritic cell TLR4 induces Th1-type immune response against Cryptosporidium parvum infection

@#The objective of this study was to investigate the mechanism of Toll-like receptor (TLR4)- mediated dendritic cell (DC) immune against Cryptosporidium parvum infection. C. parvum sporozoites were labeled with 5,6-carboxyfluorescein diacetate succinimidyl ester. Murine bone marrow-derived DCs were isolated, and divided into TLR4 antibody blocking (TAB; infected with 2 × 105 labeled sporozoites and 0.5 μg TLR4 blocking antibody), TLR4 antibody unblocking (TAU; infected with 2 × 105 labeled sporozoites), and blank control (BC; with 1.5 mL Roswell Park Memorial Institute 1640 medium) groups. The adhesion of Cryptosporidium sporozoites to DCs and CD11c+ levels were examined by fluorescence microscopy and flow cytometry. Male KM mice were orally injected with C. parvum. The proliferation of T lymphocytes in spleen, expression of cytokines in peripheral blood, and TLR4 distribution features in different organs were further determined by immunohistochemistry. A significantly higher expression of CD11c+ and higher C. parvum sporozoite adhesion were found in the TAU group compared with other groups. The expression of CD4+CD8- /CD8+CD4- in the spleen were obviously differences between the TAB and TAU groups. The expression of TLR4, interleukin IL-4, IL-12, IL-18 and IFN-γ improved in the TAU group compared with TAB group. Higher expression of TLR4 was detected in the lymph nodes of mice in the TAU group, with pathological changes in the small intestine. Hence, TLR4 could mediate DCs to recognize C. parvum, inducing Th1 immune reaction to control C. parvum infection.

[Active screening of intestinal carbapenem-resistant Enterobacteriaceae in high-risk patients admitted to the hematology wards and its effect evaluation].

Objective: To evaluate the effect of intestinal carbapenem-resistant Enterobacteriaceae (CRE) active screening combined with enhanced intervention in the prevention and control of nosocomial infection in patients admitted to the hematological ward. Methods: Patients who were admitted to the Department of Hematology in a tertiary-care general hospital from March 1, 2017 to December 31, 2019 and underwent chemotherapy or immunosuppressive therapy comprised the intervention group. They were screened for intestinal CRE at least thrice. From December 1, 2016 to February 28, 2017, patients who underwent chemotherapy or immunosuppressive therapy without active intestinal CRE screening in the Department of Hematology formed the control group. Both the patient groups were monitored for CRE infection in real time. The χ(2) test was used to compare the changes in the CRE infection rate and mortality in high-risk patients before and after the active screening. Results: During the intervention period, the CRE colonization rate of patients was 16.46% (66/401) ; in terms of disease distribution, the colonization rate of acute leukemia was the highest 23.03% (26/113) . Of the 66 colonized patients, 27 (40.9%) patients were identified as positive for CRE at the first screening, 15 (22.7%) were identified at the time of the second screening, and the remaining 24 (36.4%) were identified at the third or subsequent screening; Carbapenem-resistant Klebsiella pneumoniae (CRPK) strains were dominant among the pathogens, accounting for 54.55% (36/66) . During the active screening period, the CRE infection rate (2.49%) and mortality rate (50.00%) of high-risk patients were significantly lower than those of the controls (11.30% and 69.23%, respectively) . The pathogens of 10 CRE infection patients during the intervention period were exactly the same as the previous active screening pathogens, and the coincidence rate was 100%. Conclusion: The CRE colonization rate was the highest in patients with acute leukemia who were admitted in the hematology wards. CRPK is the main pathogen of CRE colonization, infection, and death. Increasing the frequency of screening can significantly raise the positive rate of screening, Active screening can effectively reduce the incidence and subsequent mortality of CRE in high-risk patients admitted in the hematological wards. High coincidence rate between CRE screening positive pathogens and subsequent CRE infection pathogens. Intestinal CRE screening can serve as an indicator of CRE bloodstream infection in patients with hematological diseases as well as provide information for antibiotics therapy.

Tailoring phase transformation strengthening and plasticity of nanostructured high entropy alloys.

Metastable high entropy alloys (HEAs) have attracted extensive attention due to their excellent combination of high strength and great plasticity. In this work, we utilize constraining effects to tailor phase transformation strengthening and plasticity of nanostructured HEA ([double bond, length as m-dash]FeCoCrNi) thin films prepared by the magnetron sputtering technique via HEA/Cu and HEA/Ni nanotwinned nanolaminates (NTNLs). It is uncovered that the HEA/Cu NTNLs without phase transformation exhibit the fashion of "smaller is stronger" and HEA layers become more favorable to detwin at a smaller layer thickness (h). By contrast, the HEA/Ni NTNLs manifest an ultra-high peak hardness plateau accompanied by the FCC-to-HCP phase transformation in HEA layers at large h≥ 25 nm, whereas they manifest size-dependent hardness when detwinning occurs at smaller h. This unusual plastic deformation behavior of HEA/X (X = Cu, Ni) NTNLs was rationalized by partial-based mechanisms. These findings open a new avenue to achieve superior mechanical properties of HEAs particularly at the nanoscale.

Phylogenetic placement of Paratrichaptum and reconsideration of Gloeophyllales.

Paratrichaptum accuratum is a large conspicuous polypore fungus growing on dead or living angiosperm trees in subtropical-boreal areas of China, Indonesia, Japan, and Taiwan. The present study places P. accuratum in the family Gloeophyllaceae that belongs to the order Gloeophyllales within Agaricomycetes (Basidiomycota), based on evidence derived from morphological and ecological characteristics, and phylogenetic analyses of sequences of nuclear rDNA regions (5.8S, nuc 18S, nuc 28S) and protein-coding genes (rpb1, rpb2, and tef1). The analyses presented in this study also give strong support for including Jaapia in Gloeophyllaceae and Gloeophyllales. Thus, the names Jaapiaceae and Jaapiales are considered here as synonyms of Gloeophyllaceae and Gloeophyllales. Since Paratrichaptum represents the earliest diverging lineage in Gloeophyllales, pileate basidiocarps and brown rot appear to be ancestral states of Gloeophyllales. Paratrichaptum accuratum may represent a relic species, according to its phylogenetic position, peculiar distribution pattern and rare occurrence.

[Effect of human Wharton's Jelly or adipose derived mesenchymal stem cells culture supernatant on endothelial cells angiogenesis].

Objective: To compare the curative effect of mesenchymal stem cells derived from human Wharton's Jelly(WJ-MSC) or adipose(AD-MSC) culture supernatant on endothelial cells angiogenesis. Methods: WJ-MSC and AD-MSC were isolated, identified, and the culture supernatant of stem cells was collected.The WJ-MSC or AD-MSC supernatant co-cultured with the endothelial cells. The expression levels of pro-angiogenic and anti-angiogenic genes of endothelial cells were assessed using qRT-PCR analysis, and the effects of stem cell culture supernatant on angiogenesis were evaluated by performing a tube formation assay in vitro. Results: After adding WJ-MSC and AD-MSC culture supernatant, the expression levels of pro-angiogenic genes in endothelial cells were upregulated, and the expression levels of anti-angiogenic genes were downregulated significantly in both experimental groups compared to the control group (P<0.01), and tube formation of endothelial cells was also significantly increased in both experimental groups as determined by the increase of the tube length ((43.2±9.2) mm vs (94.3±13.2)mm, (86.1±7.2)mm, P<0.01). Conclusion: The results showed that AD-MSC culture supernatant can promote endothelial cells angiogenesis and its curative effect is similar to that of WJ-MSC.

Expression of Raptor and Rictor and their relationships with angiogenesis in colorectal cancer.

Mammalian target of rapamycin (mTOR) has two subtypes, i.e., mTORC1 and mTORC2, which contain the Raptor and Rictor core molecules, respectively. The effect of Raptor and Rictor on hypoxia inducible factor (HIF)-1α, HIF-2α, and vascular endothelial growth factor (VEGF) in colorectal cancer (CRC) is unclear. In this work, we investigated the correlations among Raptor, Rictor, HIF-1α, HIF-2α, and VEGF expression in CRC. We subsequently analyzed the clinicopathological features of patients. Immunohistochemistry, western blot, and RT-PCR analyses were performed to detect the expression of Raptor, Rictor, HIF-1α, HIF-2α, and VEGF in 120 cases of CRC and 60 cases of normal colorectal mucosa. CD34 was used to label microvascular density (MVD), which was found to be higher in patients with positive Raptor or Rictor than in those with negative Raptor or Rictor. The positive rates of Raptor, Rictor, HIF-1α, HIF-2α, and VEGF in CRC were significantly higher than in normal colorectal mucosa. Raptor expression was positively correlated with HIF-1α and VEGF but not with HIF-2α expression. By contrast, Rictor expression was positively correlated with HIF-2α and VEGF but not with HIF-1α expression. Survival analysis further indicated that Raptor, Rictor, HIF-1α, HIF-2α, VEGF and lymph node metastasis were independent prognostic factors in CRC. To conclude, Raptor and Rictor expression was related to the initiation and development of CRC and angiogenesis in different ways. The combined detection of Raptor and Rictor is important for patients with colorectal carcinoma in prognosis and optimal therapy.

[Analysis of the correlation between radiological and audiological features of patients with enlarged vestibular aqueduct].

Objective: The radiological and audiological results of patients with hearing loss associated with enlarged vestibular aqueduct (EVA) were analyzed statistically to explore the association between them. Methods: In this retrospective study, we screened 64 patients (128 ears) with EVA diagnosed in the Department of Otorhinolaryngology, Shengjing Hospital of China Medical University from January 2012 to June 2016, who met the inclusion criteria and the exclusion criteria at the same time, including 37 males (74 ears) and 27 females (54 ears), aged from 6 months to 17 years, all of whom showed varying degrees of sensorineural hearing loss (SNHL). The imaging observations included the midpoint measurement (MP) and the operculum measurement(OP) of the temporal bone HRCT, the long signal area cross-sectional area (ES(L)), the short signal area cross-sectional area (ES(S)), as well as the largest total signal area cross-sectional area (ES(T)) of the endolymphatic sac(ES) of the cochlear MRI. The audiological observations included collecting detailed medical history and subjective and/or objective audiological examinations to determine the character and degree of hearing loss. According to the progress and changes of hearing loss, they were divided into hearing stability group (86 ears) and fluctuation/progression group (42 ears). SPSS22.0 statistical software was used to carry out statistical analysis of the imaging measurement results. Results: The sizes of MP and OP were smaller in the stable group than those in the fluctuating/progressive group, and the difference was statistically significant (P<0.05). For ES(L), ES(S) or ES(T), there was no significant difference between the stable group and the fluctuating/progressive group (P>0.05). For the 13 patients with asymmetric hearing loss, the sizes of MP, OP, ES(L), ES(S) or ES(T) were not significantly different between the mild side and the serious side (P>0.05). For the ears whose ES(L)/ES(S) was>2 or the ES(S)/ES(L) was>2, the proportion was not significantly different between the stable group and the fluctuating/progressive group (P>0.05). Conclusions: For patients with EVA,the smaller the enlarged vestibular aqueduct is,the more likely it is congenital severe sensorineural hearing loss. Patients with significantly enlarged vestibular aqueduct often exhibit volatility/progressive hearing loss. The degree of enlargement of the endolymphatic vessels and endolymphatic sac is not related to the degree of hearing loss. For EVA patients with asymmetric hearing loss, the mild or serious side of hearing loss is random, which is not affected by the degree of enlargement of the vestibular aqueduct and endolymphatic sac. The degree of hearing loss in patients with EVA is not related to the ratio of ES(L)/ES(S).

[Construction of mouse CCR3 gene RNAi lentivirus vector and its expression on mast cells].

Objective:The aim of this study is to screen the targeting chemokine receptor 3-RNA interference (CCR3-RNAi) lentiviral expression vector, infect mouse mast cells,observe the expression of this gene in mast cells and the interference efficiency of the virus vector.The pathogenesis of allergic rhinitis lays the foundation.Method:Three pairs of CCR3-shRNA sequences were constructed,and three pairs of double-stranded shRNA oligo were inserted into shRNA lentiviral vectors to construct three shRNA lentiviral recombinant plasmids.The recombinant vector and virus-packed auxiliary plasmids were co-transfected into 293T cells to obtain lentiviral plasmids.The lentiviral plasmids were then transfected into mouse bone marrow-derived mast cells in vitro and purified. The expression level of CCR3 mRNA in mast cells was verified by qRT-PCR,and the expression level of CCR3 protein in mast cells was detected by Western Blot.Result: It was confirmed by sequencing that the lentiviral vector of CCR3 shRNA was successfully constructed, transfected into 293T cells and packaged with virus. Finally the high purity PDSO19-PL-CCR3 lentiviral plasmid was obtained with a virus titer of 3.7×108TU/ml.The lentiviral plasmid was used to infect mouse mast cells.RT-PCR and Western Blot detection assay showed that CCR3shRNA reduced the expression of CCR3 gene in mouse mast cells at the level of mRNA and protein.Conclusion: The CCR3 gene RNAi lentivirus expression vector was successfully constructed.It was found that it downregulated the expression level of CCR3 gene mRNA and protein in mouse mast cells,which laid the foundation for further research on its role in the pathogenesis of allergic rhinitis.

MiR-99a inhibits cell proliferation of nasopharyngeal carcinoma by targeting mTOR and serves as a prognostic factor.

OBJECTIVE: Nasopharyngeal carcinoma is the most common head and neck tumor in Southern China and Southeast Asia, presenting high rates of local invasion and early distant metastasis. Abnormally expressed miR-99a has been discovered in many tumors, and it is involved in nasopharyngeal carcinoma as well. This study aims to explore the molecular mechanisms of miR-99a and mTOR in regulating nasopharyngeal carcinoma. PATIENTS AND METHODS: MiR-99a expression in nasopharyngeal carcinoma cells was detected by quantitative Real Time-Polymerase Chain Reaction (qRT-PCR). MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide) assay was performed for accessing cell proliferative capacity. Dual-Luciferase reporter gene assay was employed to verify the combination between miR-99a and mTOR. RESULTS: We found that miR-99a was downregulated while mTOR was upregulated in nasopharyngeal carcinoma cell lines CNE1 and SUNE1. Low expression of miR-99a or high expression of mTOR predicted poor prognosis of nasopharyngeal carcinoma. MiR-99a overexpression inhibited the proliferation of CNE1 and SUNE1 cells through targeting mTOR. CONCLUSIONS: We provided evidence that miR-99a inhibits NPC cell proliferative ability by inhibiting mTOR. The newly identified miR-99a/mTOR axis provides novel insight into the pathogenesis of NPC and represents a potential therapeutic target for NPC.

[Regulatory effect of exosomes secreted by vaginal wall fibroblasts on angiogenesis in patients with stress urinary incontinence].

Objective: Exosomes are considered to mediate intercellular communication by delivering biomolecules like mRNA, miRNA into recipient cells. The purpose of this study was to evaluate the effects of exosomes secreted by fibroblasts from women with stress urinary incontinence (SUI-EXO) on endothelial cells angiogenesis. Methods: Primary fibroblasts were acquired from periurethral vaginal wall tissues and exosomes were prepared by ultracentrifugation of fibroblasts cells conditioned medium. The expression levels of pro-angiogenic and anti-angiogenic genes were assessed using qRT-PCR analysis. Migration of endothelial cells was measured by transwell assay, and the effects of SUI-EXO on angiogenesis were evaluated by performing a tube formation assay in vitro. Results: SUI-EXO was successfully isolated from fibroblasts cells conditional medium and transferred to endothelial cells efficiently. When the endothelial cells were treated with SUI-EXO, the expression levels of pro-angiogenic genes in fibroblasts were downregulated, and the expression levels of anti-angiogenic genes were upregulated significantly (P<0.01). Endothelial cells exhibited a decreased migratory capacity after treatment with SUI-EXO compared to exosomes from health women (64.6±8.7 vs 114.5±14.2,P<0.01), and tube formation of endothelial cells was also significantly inhibited in the SUI-EXO treated group as determined by the increase of the tube length (87.6±13.3 vs 168.5±28.3,P<0.01). Conclusion: This study suggests that SUI-EXO plays related roles in regulating endothelial cells angiogenesis and SUI-EXO maybe involve in the pathogenesis of SUI.

Downregulation of microRNA-196a attenuates ischemic brain injury in rats by directly targeting HMGA1.

OBJECTIVE: Dysfunction of the microRNA (miRNA) network is a major regulator in neurological diseases. However, little is known about the functional significance of miRs in ischemic brain injury. This study was designed to investigate the functional behaviors and regulatory mechanisms of miR-196a in ischemic brain injury. MATERIALS AND METHODS: Cerebral ischemia was induced by middle cerebral artery occlusion (MCAO) in rats. The expression levels of miR-196a and HMGA1 were detected by quantitative Real Time-Polymerase Chain Reaction (qRT-PCR), Western blotting, and/or immunocytochemistry. The role of miR-196a in cerebral infarction and brain cell apoptosis was determined by infarct volume estimation and Transferase-mediated deoxyuridine triphosphate-biotin nick end labeling (TUNEL) assay, respectively. Bioinformatics software and Luciferase analysis were used to predict and confirm the targets of miR-196a. RESULTS: The results showed that the miR-196a expression was significantly increased in rat brain after MCAO and in cultured neonatal rat cortical neurons after oxygen-glucose deprivation (OGD). Pretreatment with antagomiR-196a by intracerebroventricular markedly reduced the miR-196a mRNA and HMGA1 protein of the brain in rats subjected to MCAO. Notably, the knockdown of miR-196a could protect MCAO rats against cerebral infarction and brain cell apoptosis. In the OGD model, apoptosis and miR-196a expressions were inhibited, while High-Mobility-Group-A1 (HMGA1) levels were increased in cortical neurons in a time-dependent manner. Moreover, HMGA1 was the target gene of miR-196a. MiR-196a overexpression promoted OGD-induced cortical neurons apoptosis possibly through negative regulation of HMGA1. CONCLUSIONS: These findings indicated a crucial role of miR-196a in regulating infarct volume and neuronal cell death under cerebral ischemia, thus offering a new target for the development of therapeutic agents against ischemic brain injury.