Glucose-Insulin-Potassium Alleviates Intestinal Mucosal Barrier Injuries Involving Decreased Expression of Uncoupling Protein 2 and NLR Family-Pyrin Domain-Containing 3 Inflammasome in Polymicrobial Sepsis.

Uncoupling protein 2 (UCP2) may be critical for intestinal barrier function which may play a key role in the development of sepsis, and insulin has been reported to have anti-inflammatory effects. Male Sprague-Dawley rats were randomly allocated into five groups: control group, cecal ligation and puncture (CLP) group, sham surgery group, CLP plus glucose-insulin-potassium (GIK) group, and CLP plus glucose and potassium (GK) group. Ileum tissues were collected at 24 h after surgery. Histological and cytokine analyses, intestinal permeability tests, and western blots of intestinal epithelial tight junction component proteins and UCP2 were performed. Compared with CLP group, the CLP + GIK group had milder histological damage, lower levels of cytokines in the serum and ileum tissue samples, and lower UCP2 expression, whereas the CLP + GK group had no such effects. Moreover, the CLP + GIK group exhibited decreased epithelial permeability of the ileum and increased expression of zonula occludens-1, occludin, and claudin-1 in the ileum. The findings demonstrated that the UCP2 and NLR family-pyrin domain-containing 3/caspase 1/interleukin 1ß signaling pathway may be involved in intestinal barrier injury and that GIK treatment decreased intestinal barrier permeability. Thus, GIK may be a useful treatment for intestinal barrier injury during sepsis.

The TLR3, PI3K, survivin, FasL, and Fas genes as major risk factors of occurrence and development of cervical cancer disease.

To investigate the role of TLR3/PI3K signals in the occurrence and development of cervical cancer disease, TLR3-siRNA was used to block key signaling pathways involved in cervical cancer metastasis that are pivotal to metastatic tumor cells but not to normal cells under ordinary physiologic conditions. Results show that tumor U14 cell growth, migration and invasion in TLR3-siRNA treatment group were significantly decreased. Through LY294002 suppressing targeted gene, the LY294002 treatment specifically and significantly knocked down the expressions of tumor TLR3 and PI3K proteins in cervical cancer mice. Furthermore, expressions of tumor Survivin and FasL proteins were markedly suppressed, whereas expressions of Fas protein were upregulated in LY294002 treatment group mice. LY294002 treatment suppressed tumor growth and increased the thymus and spleen indeces and survival days of cervical cancer mice. This study demonstrates that TLR3-siRNA and LY294002 treatments can markedly suppress cervical cancer cell invasion and tumor growth and increase survival life by silencing targeted genes.

[Clinical study on echocardiography combined with end-tidal CO2 monitoring in preventing the venous air embolism during operative hysteroscopy].

OBJECTIVE: To explore the value of transthoracic echocardiography(TTE) combined with partial pressure of end-tidal CO2 (P ETCO2) monitoring in preventing venous air embolism(VAE) during hysteroscopy operation. METHODS: From August 2010 to August 2012, 300 cases undergoing hysteroscopic surgery under general anaesthesia were enrolled in this study. With the echocardiography combined with P ETCO2 monitoring during hysteroscopic surgery, the time of VAE appearing in the right atrium , the extent of air embolism(grade 0-4), the changes of P ETCO2, peripheral oxygen saturation (SpO2), blood pressure (Bp), the amount of fluid intravasation and the blood loss were recorded in detail . Take the 24 cases that the grades of VAE in grade 3 or above and a decrease in P ETCO2 ≥ 5 mm Hg (1 mm Hg = 0.133 kPa) as intervention group. Stop operation immediately, the left side and raise your right shoulder, oxygen inhalation and dexamethasone 10 mg intravenous drip were given to the intervention group; the control group (5 cases in grade 3 or above and a decrease in P ETCO2<5 mm Hg) only stop operation immediately, observation patient's condition closely. RESULTS: (1) The occurrence of VAE: air embolism occurred in 34 patients among all 300 patients, all of 34 patients had evidence of gas embolism in the inferior vena cava, right atrium(RA) and right ventricle chamber, and 29 patients had evidence of continuous groups of gas emboli. There were 32 cases with the decrease in P ETCO2 > 2 mm Hg, 14 cases with SpO2 less than 95%, and 4 cases with a drop in Bp ≥ 20%. (2) The change of the decrease of P ETCO2 and the grades of VAE: the decrease of P ETCO2 and the grades of VAE were positively correlated (r = 0.601, P < 0.01), continuous groups of gas emboli signal in RA, when the drop of P ETCO2 ≥ 5 mm Hg, 8 of the 34 cases whose drop of P ETCO2 ≥ 10 mm Hg, 7 cases in grades 4. The decrease of P ETCO2 [(7.5 ± 2.4) mm Hg versus (11.1 ± 4.1) mm Hg], the amount of fluid intravasation[(688 ± 150) ml versus (925 ± 268) ml] and the blood loss[(71 ± 36) ml versus (127 ± 56) ml] all had statistical difference in grade 3 and 4 (P < 0.05) . The sensitivity of P ETCO2 was better than SpO2 and Bp in detecting of VAE. (3) The cases of P ETCO2 continue to decline and recovery time between the two groups:2 cases' P ETCO2 continued to decline in the intervention group, but 4 cases in the control group. The recovery time of the two groups was (4.8 ± 1.6) and (8.3 ± 1.9) min, respectively, which reached statistical difference between (P < 0.05). CONCLUSIONS: TTE combined with P ETCO2 monitoring can effectively predict the occurrence of air embolism in hysteroscopic surgery; Continuous groups of gas emboli signal in RA accompanied by a decrease in P ETCO2 ≥ 5 mm Hg indicates that the occurrence of air embolism; the more the amount of intravasation of distension fluid and the blood loss, the more the air into the body. Timely intervention can effectively prevent the development of air embolism when the VAE in grade 3 or above accompanied by a decrease in P ETCO2 ≥ 5 mm Hg.

[Association of methionine synthase reductase gene polymorphism with unexplained recurrent spontaneous abortion].

OBJECTIVE: To explore the relationship between the polymorphism of methionine synthase reductase (MTRR) A66G and the susceptibility to unexplained repeated spontaneous abortion (URSA). METHODS: Total of 200 Henan Han couples with URSA (URSA group) and 76 Henan Han healthy couples without URSA (control group) were enrolled in this study. Their MTRR A66G genotypes were determined by PCR restriction fragment length polymorphism (PCR-RFLP). RESULTS: (1) The allele frequencies of MTRR A66G: the frequencies of allele A and allele G in URSA group were 76.5% (153/200) in husband and 72.8% (146/200) in wife, 23.5% (47/200) in husband and 27.2% (54/200) in wife, respectively. The frequencies of allele A and allele G in control group were 78.9% (60/76) in husband and 78.3% (59/76) in wife, 21.1% (16/76) in husband and 21.7% (16/76) in wife, respectively. The frequencies of allele A and allele G were not significantly different between female and male subjects within the same experimental group (P > 0.05), and also there were not significantly different between the same gender subjects at URAS and control groups (P > 0.05). (2) The genotype frequencies of MTRR A66G: the frequencies of genotype AA, AG and GG in URSA group were 57.0% (114/200) in husband and 52.0% (104/200) in wife, 39.0% (78/200) in husband and 41.5% (83/200) in wife, 4.0% (8/200) in husband and 6.5% (13/200) in wife, prepectively. The frequencies of genotype AA, AG and GG in control group were 59.2% (45/76) in husband and 59.2% (50/76) in wife, 39.5% (30/76) in husband and 38.2% (29/76) in wife; 1.3% (1/76) in husband and 2.6% (2/76) in wife, prepectively. The frequencies of genotype AA, AG and GG were not significantly different between female and male subjects within the same group (P > 0.05), and also there were not significantly different between the same gender subjects at URSA and control groups (P > 0.05).(3)Combined genotype of couples: the combined genotype frequencies of GG + GG, GG + AG, GG + AA, AG + AG, AG + AA and AA + AA in URSA group were 1.0% (2/200), 2.5% (5/200), 6.0% (12/200), 20.0% (40/200), 38.0% (76/200), and 32.5% (65/200), prepectively; the combined genotype frequencies in control group were 0, 1.3% (1/76), 2.6% (2/76), 17.1% (13/76), 42.1% (32/76), 36.8% (28/76), prepectively. The combined genotype analysis between the two groups were also not significantly different (P > 0.05). CONCLUSION: The polymorphism of MTRR A66G gene was not associated with the susceptibility to URSA (P > 0.05), and so it was not the inherited genetic risk factor of URSA.

[Clinical application of noninvasive prenatal diagnosis using cell free fetal DNA in maternal plasma].

OBJECTIVE: To investigate the clinical value of non-invasive prenatal diagnosis using cell free fetal DNA (cff-DNA) in maternal blood. METHODS: From Sep. 2010 to Mar. 2012, 103 pregnant women who came to Henan Province People's Hospital in the first trimester for prenatal diagnosis of sex-linked inherited diseases were included in the first trimester group. From Oct. 2010 to Jan. 2012, 205 pregnant women undergoing amniotic fluid sampling for fetal karyotype analysis in the same hospital were included in the second trimester group. Real time quantitative PCR and fluorescent PCR were used to detect sex determining region of Y chromosome gene (SRY) and amelogenin gene (AML) on cff-DNA of the first trimester group. Moreover, 12 Y chromosome STR loci analysis were performed for 33 male fetuses and their fathers. Massively Parallel Signature Sequencing (MPSS) was used for aneuploidy analysis in cff-DNA of the second trimester group. RESULTS: (1) In the first trimester group, there were 53 SRY positive and 50 SRY negative. Compared with the results of cff-DNA of chorionic villus samples, there was one SRY false positive and one false negative results, with a sensitivity of 98% and specificity of 98%. For the AML gene test, there were two PCR products of male fetuses:102 bp fragment originating from X chromosome (AML X) and 108 bp fragment from Y chromosome (AML Y); but only AML X was found in products from female fetuses. In the first trimester group, 102 bp and 108 bp fragments were detected in 52 cases, and only 102 bp fragment was found in the other cases. Compared to AML results from chorionic villus samples, there were 2 false negative results, with a sensitivity of 96% and specificity of 100%. (2) For cff-DNA with plasma SRY over 30 copy/ml, Y STR loci were analyzed on cff-DNA of 33 fetuses and their fathers. The Y STR loci less then 200 bp were successfully detected, while Y STR loci with PCR products between 200-300 bp showed low signal or could not be amplicated; and no PCR products more than 300 bp were detected from cff-DNA. Comparing the detected Y STR loci of cff-DNA to the fathers, 32 fetuses were concordant with their fathers'. Exogenous contamination was found in the rest one sample. (3) In the second trimester group, 6 fetuses with abnormal karyotype (two trisomy 21, three trisomy 18 and one 45, XO) were detected by cff-DNA and were proved by karyotype analysis. Moreover, the MPSS results of cff-DNA revealed one 45, Y and one trisomy 16 whose karyotype analysis showed normal results. And in one case, MPSS suggested less chrX or chrY, that was proved to be 47, XYY by karyotype analysis. CONCLUSIONS: (1) Cff-DNA in maternal blood can be used to determine fetal gender in early prenancy with considerable sensitivity and specificity. But the trace cff-DNA and the high maternal DNA background might have impact on the result. (2) Analysis of cff-DNA in maternal blood of the second trimester women showed that MPSS could be used for prenatal screening of trisomy 21 and trisomy 18. However, further research should be done for other chromosomes aneuploidy detection.