福尔马林固定石蜡包埋组织DNA提取方法对PCR的影响

目的：比较多种福尔马林固定石蜡包埋组织ＤＮＡ提取方法对ＰＣＲ的影响,探讨临床组织病理标本理想的ＤＮＡ提取方法及ＰＣＲ应用。方法：采用５种不同的ＤＮＡ提取方法应用ＰＣＲ技术进行了比较。结果：Ｔｗｅｅｎ ２０,ＮＰ ４０加蛋白酶Ｋ法和亚氨基二乙酸法处理标本时ＰＣＲ结果较为理想。结论：合适的福尔马林固定石蜡包埋组织ＤＮＡ提取方法可应用ＰＣＲ进行ＤＮＡ水平的相应的研究。     

Changes and significance of plasma fibrinogen gamma‐chain concentration in preeclampsia patients

ObjectiveTo investigate the plasma fibrinogen gamma‐chain concentration in preeclampsia patients and explore its value in preeclampsia prediction and auxiliary diagnosis.MethodsFollow‐up of pregnant women who regularly attended perinatal care at two hospitals in China was performed, and clinical data and plasma samples were collected at each examination until delivery. The gamma‐chain concentration was detected by Western blotting, and Quantity One Software was used for gamma‐chain grayscale value measurements.ResultsForty‐two patients with preeclampsia and 42 control patients completed the follow‐up. In the control group, the gamma‐chain concentration at 32 weeks of gestation was higher than that at 20 weeks of gestation, but the difference was not statistically significant (p > 0.05). In the experimental group, the gamma‐chain concentration at preeclampsia diagnosis was significantly higher than that at 20 weeks of gestation (p < 0.05). Compared with the control group, the gamma‐chain concentration was higher at 20 weeks of gestation in the experimental group, but the difference was not statistically significant. However, at 32 weeks of gestation and at the time of diagnosis, the gamma‐chain concentration in the experimental group was significantly higher than that in the control group (p < 0.05). At 32 weeks of gestation and at the time of diagnosis, the AUCs from ROC curve analysis of plasma fibrinogen gamma‐chain concentrations were 0.64 and 0.71, respectively.ConclusionPlasma fibrinogen synthesis and degradation were disrupted in preeclampsia patients before and after diagnosis, and gamma‐chain concentration was significantly increased. Plasma fibrinogen gamma chain may be of some value in preeclampsia prediction and auxiliary diagnosis.     

Flake-like InVO4 modified TiO2 nanofibers with longer carrier lifetimes for visible-light photocatalysts

Highly efficient solar light absorption capabilities and quantum yields in photocatalysts are key to their application in photocatalytic fields. Towards this end, TiO₂/InVO₄ nanofibers (NFs) have been designed and fabricated successfully by a one-pot electrospinning process. The resulting TiO₂/InVO₄ NFs display excellent visible-light photocatalytic activity, owing to their prominent visible-light absorption and electron–hole separation properties. Time-resolved transient PL spectroscopy demonstrated that the TiO₂/InVO₄ NFs display longer emission decay times (22.0 ns) compared with TiO₂ NFs (15.5 ns), implying that the heterojunction can remarkably suppress the electron–hole recombination and promote the carrier transfer efficiency. With tailored heterostructure features, TiO₂/InVO₄ NFs exhibit superior visible-light photodegradation activity, and after 80 min of visible-light irradiation, almost 95% of RhB molecules can be decomposed by TiO₂/InVO₄ NFs, while only 18% of RhB molecules can be decomposed by pure TiO₂ NFs.

TiO₂/InVO₄ nanofibers have been designed and fabricated successfully by one-pot electrospinning process, which display longer carrier lifetime (22 ns) and enhanced visible-light photocatalytic activity.     

Pharmacokinetics of Pazufloxacin Mesilate Sodium Chloride in healthy Chinese volunteers after intravenous injection

In the present study, we aimed to investigate the pharmacokinetics and dosage proportionality for a single, intravenous utilization of Pazufloxacin Mesilate Sodium Chloride, an injectable synthetic fluoroquinolone antibacterial agent, in healthy Chinesevolunteers.In this open-labeled, three-dosage parallel study, subjects were randomized to receive a single dose of PazufloxacinMesilate at 150, 300 or 600 mg (n = 10, 10 and 10, respectively) administered as a 30-min intravenous infusion. Blood and urine samples were serially collected from 0 to 24 h after drug administration. Moreover, the sample’s drug concentrations were analyzed via validated RP-HPLC method.Subjects receiving a single dose of Pazufloxacin Mesilate 150, 300 or 600 mg were in accordance with the two compartment model. The C_max for each dosage group was 2.37±0.89, 4.27±0.74 and 10.74±4.06 mg·mL^–1, respectively; and the AUC_0→∞ was 3.24±1.2, 5.89±1.51 and 13.32±2.35 mg·h·mL^–1, respectively. In addition, T_max for groups treated with 150, 300 and 600 mg was 0.48±0.08, 0.50±0.00 and 0.53±0.08 h, respectively. The correlation analysis for AUC_0→∞, C_max and dosage suggested that pazufolxacin mesilate displayed dose proportion at the dose ranging from 150 to 600 mg. The data suggested that all three different dosage regimens fit with the two compartment model. Meanwhile, it presented a linear correlation between AUC_0→∞, C_max and dosage over the range of 150–600 mg.     

Pronuclear removal of tripronuclear zygotes can establish heteroparental normal karyotypic human embryonic stem cells

Purpose

This study aimed to derive heteroparental normal karyotypic human embryonic stem cells (hESCs) from microsurgically corrected tripronuclear (3PN) zygotes.

Methods

After sequential culture for 5–6 days, embryos developed from microsurgically corrected 3PN zygotes were analyzed by fluorescence in situ hybridization (FISH) using probes for chromosomes 17, X and Y. Intact 3PN zygotes from clinical in vitro fertilization (IVF) cycles were cultured as the control group. The inner cell mass (ICM) of blastocysts that developed from microsurgically corrected 3PN zygotes was used to derive hESC lines, and the stem cell characteristics of these lines were evaluated. G-banding analysis was adopted to identify the karyotype of the hESC line, and the heteroparental inheritance of the hESC line was analyzed by DNA fingerprinting analysis.

Results

The blastocyst formation rate (13.5 %) of the microsurgically corrected 3PN zygotes was significantly higher (P?<?0.05) than that of intact 3PN zygotes (8.7 %). The diploid rate of the blastocysts (55.0 %) was significantly higher (P?<?0.05) than that of the arrested cleavage-stage embryos (18.4 %) in microsurgically corrected 3PN zygotes. The triploid rate of the microsurgically corrected 3PN zygotes (5.7 %) was significantly lower (P?<?0.01) than that of intact 3PN zygotes (19.4 %). Furthermore, we established one heteroparental normal karyotypic hESC line from the microsurgically corrected tripronuclear zygotes.

Conclusions

Pronuclear removal can effectively remove the surplus chromosome set of 3PN zygotes. A combination of pronuclear removal and blastocyst culture enables the selection of diploidized blastocysts from which heteroparental normal karyotypic hESC lines can be derived.

     

The effect of concurrent nasal surgery on the eustachian tube function and myringoplasty outcomes

ObjectiveTo investigate the Effect of concurrent nasal surgery on the eustachian tube function (ETF) and myringoplasty outcomes for the chronic perforations with coexistent nasal pathology.Materials and methodsWe retrospectively reviewed the records of 93 patients with perforations who underwent same-day myringoplasty and nasal-septal surgery. Group A exhibited septal deviations (n = 34) and Group B inflammatory sinus disease (n = 59). Groups were compared with respect to pre- and postoperative air-bone gaps (ABGs), graft success rates and ETF (Eustachian tube score [ETS] and seven-item Eustachian Tube Dysfunction Questionnaire [ETDQ-7]) at 6 and 24 months.ResultsGraft success rates were 100.0% in Group A and 98.3% in Group B at 6 months postoperatively (P = 0.445). Graft success rates were 85.3% in Group A and 96.6% in Group B at 24 months postoperatively (P = 0.046), the re-perforation rate was significantly higher in Group A than in Group B (P = 0.015). Although the preoperative ETS was similar between two groups, the postoperative ETS in the Group B was significantly higher compared with Group A regardless of at postoperative 6th and 24th months. In addition, difference was significant for the patients with positive Valsalva maneuver among two groups at postoperative 24th months. Also, the improvement in the ETDQ-7 score in the B group was significantly higher than that in the A group at postoperative 6th and 24th months.ConclusionsConcurrent nasal surgery and myringoplasty is feasible. In addition, ESS improves ETF and thus long-term outcomes of myringoplasty for the chronic perforations with inflammatory sinus disease.     

Inflammatory Markers and Risk of Type 2 Diabetes: A systematic review and meta-analysis

OBJECTIVE

There has been growing evidence that inflammatory markers play a role in the development of type 2 diabetes. We aimed to systematically review prospective studies on the associations of elevated levels of interleukin-6 (IL-6) and C-reactive protein (CRP) with increased risk of type 2 diabetes by conducting a meta-analysis.

RESEARCH DESIGN AND METHODS

A systematic search of the PubMed, EMBASE, ISI Web of Knowledge, and Cochrane Library databases up until 10 February 2012 was conducted to retrieve prospective studies matched to search terms. We used generalized least-squares trend estimation to assess dose-response relationships. The summary risk estimates were pooled using either fixed-effects or random-effects models to incorporate between-study variation.

RESULTS

The meta-analysis, including 10 prospective studies, with a total of 19,709 participants and 4,480 cases, detected a significant dose-response association of IL-6 levels with type 2 diabetes risk (relative risk [RR] 1.31 [95% CI 1.17–1.46]). For CRP, the meta-analysis involving 22 cohorts, with a total of 40,735 participants and 5,753 cases, showed that elevated CRP levels were significantly associated with increased risk of type 2 diabetes (1.26 [1.16–1.37]), with the absence of publication bias. Sensitivity and subgroup analyses further supported the associations.

CONCLUSIONS

This meta-analysis provides further evidence that elevated levels of IL-6 and CRP are significantly associated with increased risk of type 2 diabetes.The rapid worldwide increase in the prevalence of type 2 diabetes has become a serious public health problem (1). Type 2 diabetes may be accompanied by long-term microvascular and macrovascular complications, which lead to both morbidity and mortality (2). In addition, as many as one-third of individuals with type 2 diabetes are undiagnosed. However, accumulating evidence shows that inflammation may play a crucial intermediary role in the pathogenesis of type 2 diabetes, thus relating diabetes to a number of commonly coexisting conditions thought to originate via inflammatory mechanisms (3). In this regard, more recent data suggest that interleukin-6 (IL-6) and C-reactive protein (CRP) are associated with type 2 diabetes (4–10). IL-6, a pleiotropic proinflammatory cytokine, is produced by a variety of cells, including activated leukocytes, endothelial cells, and adipocytes (11). CRP is an acute-phase plasma protein synthesized by the liver and has been shown to be a sensitive, systemic biomarker of inflammation (3). The stability of this protein during long-term frozen blood storage and the availability of inexpensive, precise, and standardized assays have assisted studies of CRP (12).One potential implication of the many studies suggesting a relation between inflammation and diabetes is that inflammatory markers may be used to refine diabetes risk prediction and thus better target individuals for lifestyle interventions. However, the results reported on the association between IL-6 and diabetes risk have varied across studies (13–16). To date, no systematic review has been performed to evaluate the available evidence on the association of IL-6 levels with the risk of type 2 diabetes. Two previous meta-analyses evaluating the association of CRP and diabetes risk have yielded contradictory results. One previous meta-analysis (17) suggested that a positive association exists between CRP and diabetes risk. In contrast, another meta-analysis (18) concluded that CRP may not be an independent risk factor for the development of diabetes.The objective of the current study was to estimate the magnitude of the relationships between IL-6 and CRP levels and the risk of type 2 diabetes in prospective studies and to quantify these relationships in a meta-analysis.     

Studies on interaction between Vitamin B12 and human serum albumin

The binding reaction between Vitamin B12 (B12, cyanocobalamin) and human serum albumin (HSA) was investigated by fluorescence quenching, UV–vis absorption and circular dichroism (CD) spectroscopy. Under simulative physiological conditions, fluorescence quenching data revealed that the quenching constants (K_sv) are 3.99 × 10⁴, 4.33 × 10⁴, 4.76 × 10⁴ and 5.16 × 10⁴ M⁻¹ at 292, 298, 304 and 310 K, respectively. The number of binding sites, n is almost constant around 1.0. On the basis of the results of fluorescence quenching the mechanism of the interaction of B12 with HSA has been found to be a dynamic quenching procedure. Thermodynamic parameters ΔH^Θ = −13.38 kJ mol⁻¹, ΔS^Θ = 66.73 J mol⁻¹ K⁻¹ were calculated based on the binding constant. These suggested that the binding reaction was enthalpy and entropy driven, and the electrostatic interaction played major role in stabilizing the reversible complex. The binding distance r = 5.5 nm between HSA and B12 was obtained according to Förster theory of energy transfer. The effect of B12 on the conformation of HSA was analyzed by synchronous fluorescence and CD spectroscopy. Synchronous spectra indicated that the polarity around the tryptophan (Trp214) residues of HSA was decreased and its hydrophobicity was increased; however, the α-helix content of the protein was predominant in the secondary structure but the CD spectra indicated that B12 induced minor conformational changes of HSA.     

反相高效液相色谱法测定硫酸阿托品注射液的含量

目的建立一种用RP-HPLC法测定硫酸阿托品注射液中硫酸阿托品含量的方法。方法色谱柱LichrospherC18(200×4.6mm,5μm);流动相乙腈-0.01%庚烷磺酸钠溶液(22∶78)。流速1.0ml/min;温度25℃;检测波长208nm。结果硫酸阿托品在20.4μg/ml~102.0μg/ml范围内峰面积与浓度呈良好的线性关系,回归方程为Y=1.34×104X-2.96×104r=0.9998(n=3),回收率为100.0%,RSD为0.7%。结论本法简单,快速,可靠,重现性好。