Establishment and comparison of different procedures for modeling intrauterine adhesion in rats: A preliminary study.

The establishment of a stable animal model for intrauterine adhesion (IUA) can significantly enhance research on the pathogenesis and pathological changes of this disease, as well as on the development of innovative therapeutic approaches. In this study, three different modeling methods, including phenol mucilage combined mechanical scraping, ethanol combined mechanical scraping and ethanol modeling alone were designed. The morphological characteristics of the models were evaluated. The underlying mechanisms and fertility capacity of the ethanol modeling group were analyzed and compared to those of the sham surgery group. All three methods resulted in severe intrauterine adhesions, with ethanol being identified as a reliable modeling agent and was subsequently subjected to further evaluation. Immunohistochemistry and RT-PCR results indicated that the ethanol modeling group exhibited an increase in the degree of fibrosis and inflammation, as well as a significant reduction in endometrial thickness, gland number, vascularization, and endometrial receptivity, ultimately resulting in the loss of fertility capacity. The aforementioned findings indicate that the intrauterine perfusion of 95 % ethanol is efficacious in inducing the development of intrauterine adhesions in rats. Given its cost-effectiveness, efficacy, and stability in IUA formation, the use of 95 % ethanol intrauterine perfusion may serve as a novel platform for evaluating innovative anti-adhesion materials and bioengineered therapies.

[Holocarboxylase synthetase deficiency induced by HLCS gene mutations: a rare disease study]. / ç½•è§ç— ç ”ç©¶ï¼šHLCSåŸºå› çªå˜è‡´å ¨ç¾§åŒ–é ¶åˆæˆé ¶ç¼ºä¹ç—‡.

A boy, aged 16 months, attended the hospital due to head and facial erythema for 15 months and vulva erythema for 10 months with aggravation for 5 days. The boy developed perioral and periocular erythema in the neonatal period and had erythema and papules with desquamation and erosion in the neck, armpit, and trigone of vulva in infancy. Blood gas analysis showed metabolic acidosis; the analysis of amino acid and acylcarnitine profiles for inherited metabolic diseases and the analysis of organic acid in urine suggested multiple carboxylase deficiency; genetic testing showed a homozygous mutation of c.1522C>T(p.R508W) in the HLCS gene. Finally the boy was diagnosed with holocarboxylase synthetase deficiency and achieved a good clinical outcome after oral biotin treatment. This article analyzes the clinical data of a child with holocarboxylase synthetase deficiency and summarizes the etiology, diagnosis, and treatment of this child, so as to provide ideas for clinicians to diagnose this rare disease.

[Recurrent vulvar rashes in a girl aged 22 months for more than half a year].

A girl, aged 22 months, attended the hospital due to recurrent vulvar rashes for more than half a year. Skin biopsy showed Langerhans cell histiocytosis, and evaluation of systemic conditions showed no systemic involvement. Therefore, the girl was diagnosed with Langerhans cell histiocytosis (skin type). In conclusion, for rashes on the vulva alone, if there are no specific clinical manifestations, the possibility of Langerhans cell histiocytosis should be considered after molluscum contagiosum, sexually transmitted diseases, and Fordyce disease are excluded.

Magnolol attenuates the inflammation and apoptosis through the activation of SIRT1 in experimental stroke rats.

BACKGROUND: Silent information regulator 1 (SIRT1), a histone deacetylase, plays a protective role in ischemic brain injury. Previous studies have shown that magnolol has a beneficial effect on ischemic stroke; however, the role of SIRT1 in the protective effect of magnolol against cerebral ischemia has not been investigated. METHODS: We used a middle cerebral artery occlusion model of stroke in rats. Before stroke induction, the rats received intraperitoneal injections of magnolol with or without the SIRT1 inhibitor, EX527. Brain water content, neurological score, and infarct volume were measured. Moreover, the levels of the proinflammatory cytokines tumor necrosis factor-α (TNF-α) and interleukin-1ß (IL-1ß) were measured. Western blot analysis was performed to detect Ac-FOXO1, SIRT1, bax, and Bcl-2 expression. RESULTS: Magnolol exerted a beneficial effect on cerebral ischemia, as indicated by reduced brain edema, decreased infarct volume, and improved neurological score. Magnolol had an anti-inflammatory effect mediated by a decrease in the expression of IL-1ß and TNF-α in the brain tissue. Additionally, magnolol down-regulated bax and Ac-FOXO1 expression and up-regulated Bcl-2 and SIRT1 expression. This effect of magnolol was abolished by EX527 treatment. CONCLUSION: In conclusion, our data clearly indicate that magnolol modulates brain injury caused by ischemic stroke by inhibiting inflammatory cytokines and apoptosis through SIRT1 activation.

Construction and characterization of HIV type 1 CRF07_BC infectious molecular clone from men who have sex with men.

This study aimed to investigate the biological characterization of HIV type 1 (HIV-1) CRF07_BC infection among men who have sex with men (MSM). From November 2011 to November 2013, a total of 66 blood samples were collected from MSM with acute HIV-1 infection with CRF07_BC subgroup strains. Deletion in the gag p6 region was detected by sequence alignment and comparative analysis. Peripheral blood mononuclear cells (PBMCs) of HNXX1301-1307 samples were separated by density gradient centrifugation. Nested polymerase chain reaction (nPCR) was used to amplify the viral DNA. The near full-length HIV-1 DNA products were ligated to the long terminal repeat (LTR) vector plasmid (07BCLTR) to construct a full-length HIV clone. The molecular clone was transfected into HEK-293T cells, TZM-b1 cells and patients' PBMCs. The pregenome of an infectious molecular clone of HIV-1 (pNL4-3) was amplified, and a subclone with CRF07_BC was developed to construct the full-length chimeric molecular clone pNL4-3/07BCLTR. Detection of p24 antigen and luciferase activity was used to measure the in vitro infectivity of pNL4-3/07BCLTR. Among the 66 MSM patients infected with CRF07_BC strains, deletion mutations of the Gag P6 proteins were found in 7 of 18CRF07_BC strains; deletion mutations of 2-13 amino acids in different regions were discovered in 6 strains; and the remaining 42 strains did not show deletions. Seven strains with amino acids deficiency in the P6 protein accounted for 27% of all strains and 75% of all deletion genotype strains. A total of 186 full-length molecular clones of CRF07_BC were constructed. There were 5, 9, 10 and 11 clones of HNXX1302, HNXX1304, HNXX1305 and HNXX1306 that resulted in p24-positive supernatant when transfected into HEK-293T cells. Full-length clones of HNXX1302, HNXX1304, HNXX1305 and HNXX1306 showed slight infection in the transfected TZM-b1 cells, as judged by the fluorescence values of TZM-b1 cells 48h post-transfection. However, we were unable to transfect the patients' PMBCs with the above four clones. The phylogenetic tree of the C2V3 segment of the Env gene showed that a significant gene cluster was formed by all of the chimeric full-length HNXX1306 clones, and the bootstrap value for this cluster was 97.5%. Patients' PBMCs could be infected by 1306N6, 1306N13 and 1306N22 chimeric full-length clones. The CRF07_BC subtype (6889-7407 nucleotide residues of HXB2) is one of the most prevalent epidemic HIV-1 virus strains among the MSM population. The full-length chimeric molecular clone pNL4-3/07BCLTR may significantly improve the in vitro infectivity of the CRF07_BC strain.

Patterns of cortical activation following motor tasks and psychological-inducing movie cues in heroin users: an fMRI study.

OBJECTIVE: Drug abuse and addiction are worldwide health problems. However, few studies have used fMRI to investigate the effect of chronic heroin use on brain activation. This is a study along this line. METHOD: fMRI positive sites in the brain were recorded during different motor and sensory activities. RESULTS: Following motor activities, heroin users had more sites globally activated in the brain than in normal volunteers, with ex-heroin users being least reactive. Conversely, a "heroin puffing" movie produced more activation in ongoing-heroin and ex-heroin users than in the normal individuals, whereas a movie with explicit sexual content was less stimulatory in both groups of heroin users compared to normal individuals. CONCLUSIONS: These significant findings relative to the function of specific brain nuclei are discussed.

Overall survival benefits for combining targeted therapy as second-line treatment for advanced non-small-cell-lung cancer: a meta-analysis of published data.

BACKGROUND: Combining targeted therapy has been extensively investigated in previously treated advanced non-small-cell lung cancer (NSCLC), but it is still unclear whether combining targeted therapy might offer any benefits against standard monotherapy with erlotinib. We thus performed a meta-analysis of randomized controlled trials to compare the efficacy and safety of combining targeted therapy versus erlotinib alone as second-line treatment for advanced NSCLC. METHODS: Several databases were searched, including Pubmed, Embase and Cochrane databases. The endpoints were overall survival (OS), progression-free survival (PFS), overall response rate (ORR) and grade 3 or 4 adverse event (AEs). The pooled hazard ratio (HR) or odds ratio (OR), and 95% confidence intervals (CI) were calculated employing fixed- or random-effects models depending on the heterogeneity of the included trials. RESULTS: Eight eligible trials involved 2417 patients were ultimately identified. The intention to treatment (ITT) analysis demonstrated that combining targeted therapy significantly improved OS (HR 0.90, 95% CI: 0.82-0.99, p = 0.024), PFS (HR 0.83, 95% CI: 0.72-0.97, p = 0.018), and ORR (OR 1.35, 95% CI 1.01-1.80, P = 0.04). Sub-group analysis based on phases of trials, EGFR-status and KRAS status also showed that there was a tendency to improve PFS and OS in combining targeted therapy, except that PFS for patients with EGFR-mutation or wild type KRAS favored erlotinib monotherapy. Additionally, more incidence of grade 3 or 4 rash, fatigue and hypertension were observed in combining targeted therapy. CONCLUSIONS: With the available evidence, combining targeted therapy seems superior over erlotinib monotherapy as second-line treatment for advanced NSCLC. More studies are still needed to identify patients who will most likely benefit from the appropriate combining targeted therapy.

[Impact of fire on carbon dynamics of Larix gmelinii forest in Daxing'an Mountains of North-East China: a simulation with CENTURY model].

Fire is one of the important natural disturbances to forest ecosystem, giving strong impact on the ecosystem carbon dynamics. By using CENTURY model, this paper simulated the responses of the carbon budget of Larix gmelinii forest in Huzhong area of Daxing' an Mountains to different intensities of fire. The results indicated that after the fires happened, the soil total carbon pool of the forest had a slight increase in the first few years and then recovered gradually, while the stand biomass carbon pool increased after an initial decrease, with the recovery rate of carbon pool of the stand fine components being faster than that of the coarse components. The fluctuation of the carbon pools increased with the increase of fire intensity. After the fires, both the net primary productivity (NPP) of forest vegetation and the soil heterotrophic respiration increased after an initial decrease, but the recovery rate of the NPP was faster than that of soil heterotrophic respiration, resulting in the alternation of the stand functioned as a carbon source or sink. After light fire, the forest still functioned as a weak carbon sink, and quickly recovered as a carbon sink to the level before the fire happened. After other intensities fire, the forest functioned as a carbon source within 9-12 years, and then turned back to a carbon sink again. It was suggested that lower intensity forest fire could promote the regeneration of L. gmelinii forest, reduce the combustibles, and have no strong impact on the stand carbon budget, while higher intensity forest fire would lead to the serious loss of soil- and tree carbon sequestration, retard the recovery of the forest, and thereby, the forest would be a carbon source in a longer term.

[Surface physicochemical and fractal characteristics of sediments in desilting basin from Yellow River diversion reservoir].

Surface morphology and pore surface fractal characteristics of the sediment in the desilting basin of Queshan Reservoir were studied. Six sediment samples were collected and particle size, morphology, pore structure and fractal characteristics, surface elements distribution were analyzed as well. The objectives of this study were to investigate the reason for the differences among the pore surface fractal dimensions and fractal scales on the basis of different models, and discuss the effect of surface morphology of these sediment particles on their surface elements distribution. The results showed that these sediment particles with average diameter of 18-83 microm were mainly composed of clay, silt and fine sand. Their complex surface morphology and pore size distribution were reflected by wide range of the BET surface area (8.248-31.60 m2/g), average pore diameter (3.977-7.850 nm) and pore-size distribution (1.870-60.78 nm). Although the pore surface fractal dimensions (D(s)), based on fractal FHH or thermodynamic models, were 2.67-2.89, and their fractal scales generally ranged from several nanometers to tens of nanometers, the differences were still observed in D(s) values calculated from above two models because of inhomogeneity in surface pore size distribution. Therefore, the D(s) based on pore-size distribution were 2.12-2.60, these values close to D(s) calculated from fractal FHH models revealed that pore-size distribution could contribute significantly to D(s) calculation. In addition, the heterogeneous surface adsorption sites of these sediment particles caused by much complex surface morphology had strong influence on the each element distribution on the particle surface.

Effects of jingyuankang capsules on leukocyte level in AIDS patients.

OBJECTIVE: To observe the therapeutic effects of Jingyuankang capsules for leukopenia in AIDS patients. METHODS: In this randomized double-blind trial, 58 patients orally took Jingyuankang capsule, analog Leucogen tablet and the HAART (highly active anti-retroviral therapy) drugs, and the other 58 patients took Leucogen tablet, analog Jingyuankang capsule and the HAART drugs all for 6 months, during which the peripheral hemogram was periodically examined to observe the therapeutic effects of Jingyuankang capsule for leukopenia of the AIDS patients. RESULTS: With good therapeutic effect for leukopenia of the AIDS patients, Jingyuankang capsule can enhance leukocyte level as effective as Leucogen tablet in treating grade I and grade II leukopenia, and more effectively than Leucogen tablet in treating grade III leukopenia. No toxic side-effects and adverse reactions were found during the treatment and in the follow-up visit. CONCLUSION: Jingyuankang capsule can effectively treat leukopenia of the AIDS patients.

[Effect of tBHQ and sulforaphane on Nrf2-ARE signaling pathway of Caco2 cells].

OBJECTIVE: To investigate the effect of tBHQ and sulforaphane on the protein expression in Nrf2-ARE signaling pathway of Caco2 cells. METHODS: Human colorectal carcinoma Caco2 cells were treated with 20 micromol/L tBHQ and 5 micromol/L sulforaphane (SFN) respectively. Real time PCR, Western blotting and immunoflourescence staining (IF) were performed to measure the target gene expression. RESULTS: Nrf2, AKR1C1 and NQO1 protein expressions were increased time-dependently in Caco2 cells after treatment with tBHQ and SFN. Time-course experiments showed that tBHQ and SFN increased the accumulation of Nrf2, and concomitantly increased the protein levels of AKR1C1 and NQO1. Real-time PCR and Western blotting showed that tBHQ and SFN significantly increased the expression of Nrf2 at 8h after the treatment, and AKR1C1 and NQO1 at 16 h. Confocal microscopy technique showed that Nrf2 accumulated in the nucleus at 6-8 h after treatment with tBHQ. After 1 h treatment with tBHQ the nuclear Nrf2 maintained at elevated level for at least 4 h with tBHQ withdrawn. CONCLUSION: tBHQ and SFN induced nuclear accumulation of Nrf2 and activated Nrf2-dependent regulation of ARE-mediated gene expression in Caco2 cells. In addition, the results provide experimental evidence for choosing the dose and frequency of the inducer in cancer chemoprevention study and in developing inhibitors of Nrf2-ARE signaling pathway.

[Effect of DRB/alpha-Amanitin on localization of Nrf2 in A549 cells].

OBJECTIVE: To investigate the effects of transcriptional inhibitors 5, 6-dichloro-1-b-D-ribofuranosylbenzimidazole (DRB) and alpha-Amanitin on the localization of Nrf2 in the nucleus. METHODS: A549 cells were treated with DRB (50 mg/L) or alpha-Amanitin (2.5 mg/L)for 1 h and 6 h in serum-free medium, respectively. The expressions of Nrf2, HO-1, NQO1 and AKR1C were detected by Western blotting analysis. The localization of Nrf2 was determined by laser scanning confocal microscopy after cells were treated with either DRB or agr:-Amanitin for 1 h. RESULTS: The expressions of Nrf2 and Nrf2-ARE gene batteries HO-1, AKR1C and NQO1 were decreased after 6 h treated with either DRB or alpha-Amanitin. The expression of SC35 was up-regulated but RNA Pol II was down-regulated; Y12 and NPC did not significantly change. The localization of Nrf2 in the cell nucleus did not change significantly. CONCLUSION: DRB and alpha-Amanitin can down-regulate the expression of Nrf2 and its targeting proteins HO-1, AKR1C and NQO1, but may have no effect on the localization of Nrf2.

[Effect of Luteolin and its combination with chemotherapeutic drugs on cytotoxicity of cancer cells].

OBJECTIVE: To investigate the effect of Luteolin alone or combination with chemotherapentic drugs on the cytoxicity of cancer cells. METHODS: Cultured A549, Hela, MCF-7, AGS, MGC-803, Caco2 and HepG2 cells were treated with Luteolin or the combination of Luteolin with other chemotherapeutic agents (Bexarotene, Cisplatin and Bleomycin). Cell viability was measured by MTS assay and IC(50) was calculated. RESULTS: The IC(50) of Bexarotene to Hela cells was 2 micromol/L, but with the combination of 5 micromol/L of Luteolin that reduced to 0.2 micromol/L. However, the combination of Bexarotene and Luteolin did not show significant benefit in MGC-803, HepG2 cells, Caco2 and MCF-7 cells. The IC(50) of Cisplatin to Hela cells was over 30 micromol/L,but it decreased to 3 micromol/L in the presence of 5 micromol/L Luteolin; Luteolin also sensitized Cisplatin in MGC-803, HepG2 and A549 cells studied. The IC(50) of Bleomycin to Hela cells was over 100 micromol/L, but it was about 1 micromol/L in the presence of 5 micromol/L Luteolin. A549 cells were resistant to Bleomycin with an IC(50) of 100 micromol/L, 10 micromol/L Luteolin greatly enhanced the cytotoxicity of Bleomycin to the cells with the IC(50) of about 10 micromol/L. The inhibitions of MGC-803, HepG2, A549 and AGS cells didn't change by combination of Luteolin. CONCLUSION: Low concentration of Luteolin has little toxic effect on the cancer cell lines tested in the study, but it can sensitize chemotherapeutic drugs in various cancer cell lines.

4-[(Ethoxy-imino)(phen-yl)meth-yl]-5-methyl-2-phenyl-1H-pyrazol-3(2H)-one.

In the mol-ecule of the title compound, C(19)H(19)N(3)O(2), the central pyrazole ring makes dihedral angles of 9.89 (3) and 66.06 (5)° with the two phenyl rings, and the two phenyl rings form an angle of 74.05 (5)°. An intra-molecular C-H⋯O hydrogen bond forms a six-membered ring, producing an S(6) ring motif. In the crystal structure, inter-molecular N-H⋯O and C-H⋯O hydrogen bonds link each mol-ecule to two others, forming an infinite one-dimensional supra-molecular structure along the c axis.

Purification and characterization of antitoxic proteins from the serum of Agkistrodon halys Pallas.

In this study, the authors report the purification and characterization of antitoxic proteins from the serum of Agkistrodon halys Pallas. Two antitoxic proteins have been successfully isolated by the methods of (NH4)(2)SO(4) fractional precipitation, chromatography and preparative discontinuous polyacrylamide gel electrophoresis (PAGE). We have measured their molecular weights by Sephadex G-150 chromatography and 0.1% SDS-Tris-HCl discontinue PAGE respectively. Antitoxin I was about 138,000+/-40 Da and antitoxin II was about 76,000+/-40 Da, they are all single-chain peptides. We have measured their capacity to neutralize the toxicity of agkistrodotoxin (ATX), and their capacity to inhibit the PLA(2) activity of ATX. The results showed that antitoxin I could increase LD(50) of ATX from 0.25+/-0.05 to 0.445+/-0.13 mg/kg, decrease its PLA(2) activity from 2.36 to 1.72 microm/mg min, and antitoxin II could increase LD(50) of ATX from 0.25+/-0.05 to 0.56+/-0.12 mg/kg, decrease Phospholipase A(2) (PLA(2)) activity from 2.36 to 1.2 microm/mg min. When the natural antitoxins were mixed with different amounts of ATX and inoculated intraperitonially into eight mice, it was found that 0.5 mg antitoxin I could neutralize the toxicity of 0.4 mg ATX and 0.5 mg antitoxin II could neutralize the toxicity of 0.5 mg ATX completely. These antitoxic proteins could neutralize the toxicity of ATX completely and inhibit ATX's PLA(2) activity partially.

[Impaired microfilament cytoskeleton rearrangement in cytomegalovirus infected cells].

OBJECTIVE: To investigate the effect of cytomegalovirus (CMV) infection on actin and microfilament in human embryo fibroblast cells (HF) and its relationship with CMV replication. METHODS: Cell morphology was observed after the infection of CMV. Western-blot was used to measure the expression levels of beta-actin, G-actin and F-actin proteins. CMV immediately early antigen (CMV IE) in HF cells was analyzed by indirect immunofluorescence assay. Microfilament alteration was determined by cytoskeleton fluorescence probe. RESULT: CMV IE was demonstrated in more than 95% of HF cells after infection, which was primarily located in nucleus. The shape of HF cells changed from thin shuttle like to round and thick ball like, even escaping from wall after infection by CMV. Compared with control group, the expression of G-actin protein increased at 24 h of CMV infection (0.941 +/-0.061 compared with 0.714 +/-0.119, P <0.05), then decreased at 72 h, 96 h respectively(0.218 +/-.035, 0.230 +/-0.055 compared with 0.714 +/-0.119, P <0.05). The levels of F-actin in infected cells gradually decreased at 24 h, 72 h and 96 h compared with control HF cells (0.256 +/-0.021, 0.127 +/-0.032, 0.026 +/-0.008 compared with 0.373 +/-0.050, P<0.05). In infected HF cells, microfilaments were found ruptured, arranged turbulently. Cells fused and fluorescence density of microfilament markedly reduced. CONCLUSION: Cytomegalovirus can induce alteration of actins and microfilament, which may be associated with its infection, replication and reactivity in host cells.

Response of photosynthesis, morphology and growth of Hedysarum mongolicum seedlings to simulated precipitation change in Maowusu sandland.

Response patters were investigated for seedlings of Hedysarum mongolicum, a dominant shrub in Maowusu sandland, to the simulated precipitation change by artificially controlling water supply at four levels. Plant growth characters, in terms of branch number and length, leaf number and area, and biomass, increased while water supply increased. However, the effect of water supply on leaf photosynthetic rate was not significant. Root/shoot biomass ratio significantly decreased with the increase of water supply, which was considered adaptive distribution of biomass investments in the different water supply. Water supply obviously affected branching patter. Branch section number, branch number and length of the same section enhanced as water supply increased. Branch number and length were clearly positive correlation with total and aboveground biomass in four water supply treatments. Branch character fully showed plant growth.