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Objectives: 1. Determine the feasibility and efficiency of local magnetic targeting delivery of gadolinium (Gad) contrast to the inner ear in rodents. 2. Assess any potential ototoxicity of magnetic targeting delivery of Gad in the inner ear. 3. Study the utility of magnetic targeting delivery of Gad to visualize and quantify endolymphatic hydrops (EH) in a transgenic mouse model., Study Design: Controlled in vivo animal model study., Methods: Paramagnetic Gad was locally delivered to the inner ear using the magnetic targeting technique in both rat and mouse models. Efficiency of contrast delivery was assessed using magnetic resonance imaging (MRI). Ototoxicity of Gad was examined with histology of the cochlea and functional audiological tests. The Phex mouse model was used to study EH, hearing loss, and balance dysfunction. Magnetic targeting delivery of Gad contrast was used in the Phex mouse model to visualize the effects of EH using MRI., Results: Magnetic targeting improved the delivery of Gad to the inner ear and the technique was reproducible in both rat and mouse models. The delivery method did not result in microstructural damage or any significant hearing loss in a normal animal. Magnetic targeting of Gad in the Phex mouse model allowed detailed visualization and quantification of EH., Conclusion: This study provided the first evidence of the effectiveness and efficiency of the local magnetic targeting delivery of gadolinium contrast to the inner ear and its application to the visualization and quantification of EH. Laryngoscope, 133:914-923, 2023., (© 2022 The American Laryngological, Rhinological and Otological Society, Inc.)

Objective: The aim of this research was to determine whether systemic inflammatory indicators, including aggregate index of systemic inflammation (AISI), neutrophils lymphocyte to platelet ratio (NLPR), systemic immune-inflammation index (SII), and systemic inflammation response index (SIRI), are related to bone mineral density (BMD) in perimenopausal and postmenopausal women., Methods: One hundred and eighty-one perimenopausal and 390 postmenopausal women were enrolled in this cross-sectional study. Continuous variables by analysis of variance and Kruskal Wallis test for comparing the clinical characteristics. Linear regression analysis was conducted to investigate the associations between inflammatory indicators with BMD. The comparison between the subgroups was performed using the nonparametric test and the T -test., Results: AISI, NLPR, SII, and SIRI quartile values were inversely associated with BMD in menopausal women ( P = 0.021; P = 0.047; P < 0.001; P < 0.001, respectively). After adjusting for confounding factors, four inflammatory indicators remained significantly associated with BMD (all P for trend <0.001). Analysis according to menopausal status demonstrated that AISI, SII, and SIRI were significantly correlated with mean femoral neck BMD in postmenopausal women ( P for trend = 0.015, 0.004, and 0.001), but not significantly associated with BMD in perimenopausal women ( P for trend = 0.248, 0.054, and 0.352) after adjustment for covariates., Conclusion: The quartile values of AISI, SII, and SIRI were inversely associated with BMD in postmenopausal women, following adjustment for individual variables, hormone profiles and glucolipid metabolism profiles. AISI, SII, and SIRI have potential to be important tools for screening and prevention of bone loss in menopausal women in future clinical practice., Competing Interests: The authors report no conflicts of interest in this work., (© 2023 Chen et al.)

Objective: To investigate the efficacy of oral letrozole (LE) starting on day 3 or 5 of the menstrual cycle in patients with polycystic ovary syndrome (PCOS)., Design: Retrospective cohort study., Setting: Reproductive Endocrinology Department of Hangzhou Women's Hospital., Methods: In this retrospective analysis, we analyzed patients who received oral LE for ovulation induction (OI) at the Hangzhou Women's Hospital from January 2016 to January 2021. In total, 539 PCOS patients with fertility requirements were classified into the D3 group and D5 group according to the different starting times of oral LE, that is, from the 3rd or 5th day of the menstrual cycle or LE is taken orally for 5 days starting on day 3 or 5 of progesterone withdrawal bleeding. Treatment started with one tablet (LE 2.5 mg), continue the regimen from the previous cycle in non-responders and continued until pregnancy or for up to three ovulatory cycles, with visits to determine ovulation and pregnancy, followed by tracking of pregnancies. The primary outcome was to compare ovulation rates, conception rates, live birth rates, pregnancy complications, and pregnancy outcomes at different initiation times., Results: Women who started LE on the 5th day of their menstrual cycle had more cumulative conception rates than those who started LE on the 3rd day(173 of 228[75.9%] vs . 201 of 311[64.6%], P= 0.005; rate ratio for conception, 1.174; 95% confidence interval,1.052 to 1.311) without significant differences in overall live birth rate, though there were 142 of 228[62.3%] in the D5 group versus 172 of 311[55.3%] in the D3 group (P= 0.105). The median (IQR) endometrial thickness was significantly (P = 0.013) greater during the D5 group treatment compared to the D3 group, which may be related to higher conception and clinical pregnancy rates. The median (IQR) maximum follicle diameter was not statistically (P = 0.073) different between the two groups. The cumulative ovulation per cycle rate was higher with D5 than with D3 (287 of 405 treatment cycles [70.9%] vs . 388 of 640 treatment cycles [60.6%], P=0.001). There were no significant between-group differences in pregnancy loss (31 of 173 conceptions in the D5 group [17.9%] and 29 of 201 conceptions in the D3 group [14.4%]) or multiples pregnancy (8.2% and 10.5%, respectively). Rates of other adverse events during pregnancy were similar in the two treatment groups., Conclusion: As compared with D3 group, D5 group was associated with higher ovulation and conception rates, shorter time-to-pregnancy among infertile women with the PCOS., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2022 Shi, Ye, Gao, Chen, Jin and Zhang.)

Objective: We aimed to evaluate the effectiveness of different triage strategies for high-risk human papillomavirus (hrHPV)-positive women in primary healthcare settings in China., Methods: This study was undertaken in 11 rural and 9 urban sites. Women aged 35-64 years old were enrolled. HrHPV-positive women were randomly allocated to liquid-based cytology (LBC), visual inspection with acetic acid and Lugol's iodine (VIA/VILI) (rural only) triage, or directly referred to colposcopy (direct COLP). At 24 months, hrHPV testing, LBC and VIA/VILI were conducted for combined screening., Results: In rural sites, 1,949 hrHPV-positive women were analyzed. A total of 852, 218 and 480 women were randomly assigned to direct COLP, LBC and VIA/VILI. At baseline, colposcopy referral rates of LBC or VIA/VILI triage could be reduced by 70%-80%. LBC (n=3 and n=7) or VIA/VILI (n=8 and n=26) could significantly decrease the number of colposcopies needed to detect one cervical intraepithelial neoplasia (CIN) 2 or worse and CIN3+ compared with direct COLP (n=14 and n=23). For the 24-month cumulative detection rate of CIN2+, VIA/VILI triage was 0.50-fold compared with LBC triage and 0.46-fold with the direct COLP. When stratified by age, baseline LBC triage+ performed best (P<0.001), peaking among women aged 35-44 years (P trend =0.002). In urban sites, 1,728 women were hrHPV genotyping test positive. A total of 408, 571 and 568 women were randomly assigned to direct COLP for HPV16/18+, direct COLP for other hrHPV subtypes+, and LBC triage for other hrHPV subtypes+. LBC (n=12 and n=31) significantly decreased the number of colposcopies needed to detect one CIN2+ and CIN3+ compared with direct COLP (n=14 and n=44). HPV16/18+ increased the 24-month cumulative detection rate of CIN2+ (17.89%, P<0.001)., Conclusions: LBC triage for hrHPV-positive women in rural settings and direct COLP for HPV16/18+ women and LBC triage for other hrHPV subtype+ women in urban settings might be feasible strategies., (Copyright ©2022 Chinese Journal of Cancer Research. All rights reserved.)

Introduction: Abnormal placental trophoblast function is the main cause of missed abortion (MA). Src kinase-associated phosphoprotein 2 (SKAP2) indirectly affects actin reunion, which is significantly associated with cell migration., Methods: Twenty women with MA and 20 healthy women who underwent voluntarily induced abortion were included in this study. Immunohistochemistry, qRT-PCR, and western blotting were used to determine SKAP2, WAVE2, and ARP2 expression in the villous tissues. We investigated the effects of SKAP2 and the W336K mutant (blocked SKAP2 Src homology 3 function) on growth and migration in HTR8/SVneo cells using the CCK8 assay, flow cytometry, and transwell assay. The effects of SKAP2 on the WAVE2-ARP2/3 signaling pathway in HTR8/SVneo cells were evaluated by western blotting and immunofluorescence., Results: Compared to the women in the voluntary abortion group, SKAP2 and WAVE2 expression levels were downregulated in the villous of patients with MA. In HTR8/SVneo cells, SKAP2 siRNA silencing regulated the growth and migration, while SKAP2 overexpression promoted growth and migration, and inhibited apoptosis. Additionally, SKAP2 regulated the expression of WAVE2 and ARP2, as well as the colocalization of actin with WAVE2. The SKAP2 W336K mutant could not alter WAVE2 and ARP2 expression, nor HTR8/SVneo cell growth and migration, with or without SKAP2 siRNA transfection., Discussion: SKAP2 could activate the WAVE2-ARP2/3 pathway resulting in an increase of growth and migration in trophoblasts. SKAP2 probably played an important role in MA by affecting the growth and migration of trophoblasts., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper, (Copyright © 2022. Published by Elsevier Ltd.)

Whether the first trimester maternal mean arterial pressure (MAP), placental growth factor (PlGF), and pregnancy-associated plasma protein A (PAPP-A) can predict hypertensive disorders of pregnancy (HDP) is unclear. We conducted a retrospective case-control study with the total population of 539 gravidas, of these 447 had normal pregnancy, 27 had gestational hypertension (GH), 36 had preeclampsia (PE), and 29 had preeclampsia with severe features (SPE). Prediction for HDP was determined by the area under curve (AUC). Compared to the healthy group, the multiple of the median (MoM) for MAP was increased in the study groups, while PlGF and PAPP-A were decreased. When the cutoff values for MAP, PlGF, and PAPP-A were 1.069, 0.769, and 0.673 MoM, respectively, the sensitivities for predicting HDP were 0.517, 0.446, and 0.500 and the specificities were 0.744, 0.826, and 0.769, respectively. To predict GH, the highest AUC was 0.755 (95% CI: 0.655-0.856, p < 0.001) based on MAP, PlGF, and PAPP-A. The combined PlGF and PAPP-A had the highest AUC (0.683 [95% CI: 0.584-0.782, p < 0.001] and 0.755 [95% CI: 0.682-0.829, p < 0.001]) for prediction of PE and SPE. We found that MAP, serum levels of PlGF, and PAPP-A in the first trimester pregnancy are markers that predict HDP in the third trimester. The combination of markers is far superior to single markers alone. To improve the diagnostic value, specific cutoff values should be applied to GH, PE, SPE in each condition., (© 2021. The Author(s), under exclusive licence to Springer Nature Limited part of Springer Nature.)

Key Message: Expression of Cre recombinase by AtRps5a pro or AtDD45 pro enabled Cre/lox-mediated recombination at an early embryonic developmental stage upon crossing, activating transgenes in the hybrid cowpea and tobacco. Genetic engineering ideally results in precise spatiotemporal control of transgene expression. To activate transgenes exclusively in a hybrid upon fertilization, we evaluated a Cre/lox-mediated gene activation system with the Cre recombinase expressed by either AtRps5a or AtDD45 promoters that showed activity in egg cells and young embryos. In crosses between Cre recombinase lines and transgenic lines harboring a lox-excision reporter cassette with ZsGreen driven by the AtUbq3 promoter after Cre/lox-mediated recombination, we observed complete excision of the lox-flanked intervening DNA sequence between the AtUbq3 pro and the ZsGreen coding sequence in F 1 progeny upon genotyping but no ZsGreen expression in F 1 seeds or seedlings. The incapability to observe ZsGreen fluorescence was attributed to the activity of the AtUbq3 pro . Strong ZsGreen expression in F 1 seeds was observed after recombination when ZsGreen was driven by the AtUbq10 promoter. Using the AtDD45 pro to express Cre resulted in more variation in recombination frequencies between transgenic lines and crosses. Regardless of the promoter used to regulate Cre, mosaic F 1 progeny were rare, suggesting gene activation at an early embryo-developmental stage. Observation of ZsGreen-expressing tobacco embryos at the globular stage from crosses with the AtRps5a pro Cre lines pollinated by the AtUbq3 pro lox line supported the early activation mode., (© 2021. The Author(s).)

Background: Epithelial ovarian carcinoma (EOC) is a malignant tumor with high motility in women. Our previous study found that dysregulated nucleoside-triphosphatase cancer-related (NTPCR) was associated with the prognosis of EOC patients, and thus, this present study attempted to explore the potential roles of NTPCR in disease progression., Methods: Expressed level of NTPCR was investigated in EOC tissues by RT-qPCR and Western blot analysis. NTPCR shRNA and overexpression vector were generated and transfected into OVCAR-3 or SKOV3 cells to detect the effect of NTPCR on cell proliferation, cell cycle, cell migration, and invasion. Transcriptomic sequencing and metabolite profiling analysis were performed in shNTPCR groups to identify transcriptome or metabolite alteration that might contribute to EOC. Finally, we searched the overlapped signaling pathways correlated with differential metabolites and differentially expressed genes (DEGs) by integrating analysis., Results: Comparing para-cancerous tissues, we found that NTPCR is highly expressed in cancer tissues ( p < 0.05). Overexpression of NTPCR inhibited cell proliferation, migration, and invasion and reduced the proportion of S- and G2/M-phase cells, while downregulation of NTPCR showed the opposite results. RNA sequencing analysis demonstrated cohorts of DEGs were identified in shNTPCR samples. Protein-protein interaction networks were constructed for DEGs. STAT1 (degree = 43) and OAS2 (degree = 36) were identified as hub genes in the network. Several miRNAs together with target genes were predicted to be crucial genes related to disease progression, including hsa-miR-124-3p , hsa-miR-30a-5p , hsa-miR-146a-5 , EP300 , GATA2 , and STAT3 . We also screened the differential metabolites from shNTPCR samples, including 22 upregulated and 22 downregulated metabolites. By integrating transcriptomics and metabolomics analysis, eight overlapped pathways were correlated with these DEGs and differential metabolites, such as primary bile acid biosynthesis, protein digestion, and absorption, pentose, and glucuronate interconversions., Conclusion: NTPCR might serve as a tumor suppressor in EOC progression. Our results demonstrated that DEGs and differential metabolites were mainly related to several signaling pathways, which might be a crucial role in the progression of NTPCR regulation of EOC., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2021 Shang, Zhang, Ren, Zhao, Zhang and Tong.)

Long non-coding RNAs (lncRNAs) play crucial roles in ovarian cancer (OC) development. However, prognosis-associated lncRNAs (PALs) for OC have not been completely elucidated. Our study aimed to identify the PAL signature of OC. A total of 663 differentially expressed lncRNAs were identified in the databases. According to the weighted gene coexpression analysis, the highly correlated genes were clustered into seven modules related to the clinical phenotype of OC. A total of 25 lncRNAs that were significantly related to overall survival were screened based on univariate Cox regression analysis. The prognostic risk model constructed contained seven PALs based on the parameter λ min , which could stratify OC patients into two risk groups. The results showed that the risk groups had different overall survival rates in both The Cancer Genome Atlas (TCGA) and two verified Gene Expression Omnibus (GEO) databases. Univariate and multivariate Cox regression analyses confirmed that the risk model was an independent risk factor for OC. Gene enrichment analysis revealed that the identified genes were involved in some pathways of malignancy. The competitive endogenous RNA (ceRNA) network included five PALs, of which four were selected for cell function assays. The four PALs were downregulated in 33 collected OC tissues and 3 OC cell lines relative to the control. They were shown to regulate the proliferative, migratory, and invasive potential of OC cells via Cell Counting Kit-8 (CCK-8) and transwell assays. Our study fills the gaps of the four PALs in OC, which are worthy of further study., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2021 Zheng, Guo, Zhang, Cao, Xu, Zhang and Tong.)

Bearing design, production and complicated operating condition can lead to the scattered life cycle degradation distribution, which will bring a challenge of generalization for performance degradation assessment models. And it is costly and time-consuming to collect a large amount of labeled data for supervised diagnosis, especially when the task comes from a new operating condition. Thus in this paper, a novel bearing degradation assessment model is proposed based on transfer learning and deep hierarchical features extraction. The research of degradation assessment is transformed to the classification task of degradation pattern, which divides degradation process into the normal, slight fault, fault development and damage patterns. The hierarchical network with random weight parameters is introduced to extract the local sub-band characteristics of spectrum, in which the multiple alternately convolution and pooling layers without supervised fine-tuning are employed. Joint Geometrical and Statistical Alignment method is then utilized to obtain projected sharing feature space, and thus the knowledge of bearing degradation process is transferred to accomplish degradation pattern assessment under different operating conditions. Results of the experiments on bearing fault severity and degradation process show that the proposed method reduces the feature distribution divergence between the degradation processes and accomplishes bearing performance degradation assessment in different operating condition., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2020. Published by Elsevier Ltd.)

Introduction: A cesarean scar pregnancy (CSP), when combined with an arteriovenous malformation (AVM), is a rare, but potentially life-threatening condition that may be associated with uncontrolled hemorrhage. Hysterectomy is indicated when conservative treatment fails. Preservation of fertility is challenging., Patient Concerns: We reported a 33-year-old woman with a CSP combined with an AVM who failed methotrexate administration as conservative treatment., Diagnoses: A CSP combined with an AVM was diagnosed via three-dimensional color Doppler angiogram and magnetic resonance imaging., Interventions: Transvaginal removal of the ectopic gestation and repair of the uterine defect was performed without incident., Outcomes: The fertility of the patient was preserved and hysterectomy was avoided., Conclusion: Transvaginal fertility-sparing surgery may be successfully performed to prevent hysterectomy when conservative treatment fails in patients with a CSP combined with an AVM.

Clostridium difficile infection is the leading cause of hospital-acquired diarrhea and is mediated by the actions of two toxins, TcdA and TcdB. The toxins perturb host cell function through a multistep process of receptor binding, endocytosis, low pH-induced pore formation, and the translocation and delivery of an N-terminal glucosyltransferase domain that inactivates host GTPases. Infection studies with isogenic strains having defined toxin deletions have established TcdB as an important target for therapeutic development. Monoclonal antibodies that neutralize TcdB function have been shown to protect against C. difficile infection in animal models and reduce recurrence in humans. Here, we report the mechanism of TcdB neutralization by PA41, a humanized monoclonal antibody capable of neutralizing TcdB from a diverse array of C. difficile strains. Through a combination of structural, biochemical, and cell functional studies, involving X-ray crystallography and EM, we show that PA41 recognizes a single, highly conserved epitope on the TcdB glucosyltransferase domain and blocks productive translocation and delivery of the enzymatic cargo into the host cell. Our study reveals a unique mechanism of C. difficile toxin neutralization by a monoclonal antibody, which involves targeting a process that is conserved across the large clostridial glucosylating toxins. The PA41 antibody described here provides a valuable tool for dissecting the mechanism of toxin pore formation and translocation across the endosomal membrane.

Background: In renal diseases, earlier injury to glomerular may lead to the abscission of podocyte. The number of podocyte in urine may reflect the severity of glomerular damage. Podocalyxin (PCX) was considered as a podocyte marker. Many methods were used to detect podocyte. Applications of these methods were limited by tricky, expensive, and low accuracy. Here, we improved an immunocytochemical method to count the number of podocyte in urine., Methods: In this study, we counted the numbers of podocyte in urine by our improved method and detected the PCX levels in urine by enzyme-linked immunosorbent assay (ELISA) in glomerulopathy patients and healthy controls. The serum levels of cystatin C (CysC), blood urea nitrogen (BUN), creatinine (CR), uric acid (URIC), and β2-Microglobulin (β2-MG) in all subjects were detected. Correlation analysis and diagnostic efficiencies comparisons among immuocytochemical method, ELISA, and the biochemistry index were also performed., Results: The podocyte counts in patients were significantly higher than controls. Podocytes counts positively correlated with PCX concentrations and the serum CysC. The podocyte count had higher diagnostic efficiency than PCX concentrations detected by ELISA and serum CysC., Conclusions: The podocyte count detected by our improved method had higher diagnostic efficiency than ELISA and serum CysC., (© 2016 Wiley Periodicals, Inc.)

The Clostridium difficile toxin B is one of the main virulence factors and plays an important role in the pathogenesis of C. difficile infection (CDI). We recently revealed crucial residues in the translocation domain of TcdB for the pore formation and toxin translocation. In this study, we investigated the effects of mutating a critical site involved in pore formation, Leu-1106, to residues that differ in size and polarity (Phe, Ala, Cys, Asp). We observed a broad range of effects on TcdB function in vitro consistent with the role of this site in pore formation and translocation. We show that mice challenged systemically with a lethal dose (LD100) of the most defective mutant (L1106K) showed no symptoms of disease highlighting the importance of this residue and the translocation domain in disease pathogenesis. These findings offer insights into the structure function of the toxin translocation pore, and inform novel therapeutic strategies against CDI., (© FEMS 2015. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.)

Agrobacterium -mediated plant transformation is typically conducted by inoculating plant tissues with an Agrobacterium suspension containing approximately 10 8 -10 9 bacteria mL -1 , followed by a 2-3-d co-culture period. Use of longer co-culture periods could potentially increase transformation efficiencies by allowing more time for Agrobacterium to interact with plant cells, but bacterial overgrowth is likely to occur, leading to severe tissue browning and reduced transformation and regeneration. Low bacterial inoculum levels were therefore evaluated as a means to reduce the negative outcomes associated with long co-culture. The use of low inoculum bacterial suspensions (approximately 6 × 10 2 bacteria mL -1 ) followed by long co-culture (15 d) led to the production of an average of three transformed sunflower shoots per explant while the use of high inoculum (approximately 6 × 10 8 bacteria mL -1 ) followed by short co-culture (3 d) led to no transformed shoots. Low inoculum and long co-culture acted synergistically, and both were required for the improvement of sunflower transformation. Gene expression analysis via qRT-PCR showed that genes related to plant defense response were generally expressed at lower levels in the explants treated with low inoculum than those treated with high inoculum during 15 d of co-culture, suggesting that low inoculum reduced the induction of plant defense responses. The use of low inoculum with long co-culture (LI/LC) led to large increases in sunflower transformation efficiency. This method has great potential for improving transformation efficiencies and expanding the types of target tissues amenable for transformation of different plant species.

Disease associated with Clostridium difficile infection is caused by the actions of the homologous toxins TcdA and TcdB on colonic epithelial cells. Binding to target cells triggers toxin internalization into acidified vesicles, whereupon cryptic segments from within the 1,050-aa translocation domain unfurl and insert into the bounding membrane, creating a transmembrane passageway to the cytosol. Our current understanding of the mechanisms underlying pore formation and the subsequent translocation of the upstream cytotoxic domain to the cytosol is limited by the lack of information available regarding the identity and architecture of the transmembrane pore. Here, through systematic perturbation of conserved sites within predicted membrane-insertion elements of the translocation domain, we uncovered highly sensitive residues--clustered between amino acids 1,035 and 1,107--that when individually mutated, reduced cellular toxicity by as much as >1,000-fold. We demonstrate that defective variants are defined by impaired pore formation in planar lipid bilayers and biological membranes, resulting in an inability to intoxicate cells through either apoptotic or necrotic pathways. These findings along with the unexpected similarities uncovered between the pore-forming "hotspots" of TcdB and the well-characterized α-helical diphtheria toxin translocation domain provide insights into the structure and mechanism of formation of the translocation pore for this important class of pathogenic toxins.

Objective: To study the mechanism of protective effect of Jinqiaomai (JQM) on lung injury induced by Klebsiella pneumonia in rats., Method: The model of rats with Klebsiella pneumonia was established. The male SD rats were randomly divided into control group, model group, JQM (6, 3, 1.5 g x kg(-1)) three groups, levofloxacin (25 mg x kg(-1)) group, JQM (3 g x kg(-1)) + levofloxacin (25 mg x kg(-1)) group. The contents of IL-1beta, ICAM-1 and INF-gamma in the lung tissue homogenate were measured by radio-immunoassay and Elisa. TLR2/4 mRNA and MyD88 mRNA expression were detected by RT-PCR. IkappaB-alpha expression was detected by Western Blot., Result: The rats of model group had obvious lung injury, but those of JQM, JQM + levofloxacin and levofloxacin groups had less injury. The contents of IL-1beta, ICAM-1 and INF-gamma in lung tissue homogenate and the expressions of TLR2/4 mRNA, MyD88 mRNA and IkappaB-alpha in lung of model group were significantly higher than those in the control group (P < 0.05 or P < 0.01), while IL-1beta, ICAM-1 and INF-gamma of JQM groups were significantly lower than those of model group (P < 0.05 or P < 0.01). The expressions of TLR2/4 mRNA, MyD88 mRNA and IkappaB-alpha of JQM (6.3 g x kg(-1)) groups were significantly lower than those of model group(P <0. 05 or P <0. 01)., Conclusion: The lung injury induced by Klebsiella pneumonia is related to TLR2/4, MyD88 mRNA and IkappaB-alpha. To decrease the excessive expression of TLR2/4, MyD88 mRNA and IkappaB-alpha might be the main mechanism of protective effect of Jinqiaomai on lung injury.

Objective: To detect and localize the expression of macrophage colony-stimulating factor (M-CSF) in human luteinized granulosa cells and investigate the effects of follicle stimulating hormone (FSH) on M-CSF production by human luteinized granulosa cells in vitro., Methods: Human luteinized granulosa cells were isolated from follicular fluid of superovulated infertile patients undergoing intracytoplasmatic sperm injection. Some of the luteinized granulosa cells were used for detecting M-CSF by immunocytochemical staining (ABC method). Most of them were cultured with HAM's F-10 medium alone or plus various concentrations of FSH (2, 5, 15, 25 IU/ml). The media were collected after 72 hours and their M-CSF contents were measured by a solid phase enzyme-linked immunosorbant assay., Results: About 80% of the luteinized granulosa cells were positively stained for M-CSF antibody. The immunoreactive signals of M-CSF were specially located in the cytoplasma of the luteinized granulosa cells. No immunoreactivity of M-CSF was observed in vaginal squamous epithelial cells through contaminations during transvaginal oocyte retrieval. The concentration of M-CSF in cultured luteinized granulosa cells medium without FSH stimulation was low. However, addition of 2, 5, 15, 25 IU/ml concentrations of FSH caused significant dose-dependent increases of M-CSF production after 72 hours culture by luteinized granulosa cells (P < 0.01)., Conclusions: Human luteinized granulosa cells can synthesize and secrete M-CSF. FSH can directly stimulate M-CSF secretion by human luteinized granulosa cells. The mechanism of FSH regulating follicular development may partially via M-CSF/receptor pathway.

Objective: To investigate the effects of macrophage colony-stimulating factor (M-CSF) on estradiol and progesterone production by human luteinized granulosa cells in vitro and to detect M-CSF receptor in human granulosa cells., Methods: Human luteinized granulosa cells (LGC) were isolated from follicular fluid of superovulated infertile patients undergoing intracytoplasmatic sperm injection. Some of the LGC were used for detecting M-CSF receptor by immunocytochemical staining (ABC method). Most of them were cultured with HAM's F-10 medium plus various concentration of M-CSF (0, 10, 25, 50, 100, 250 ng/ml) in the presence or absence of follicle stimulating hormone (FSH). Media were collected at 72 hours after culture and estradiol (E(2)) and progesterone (P) in media were measured by enzyme immunoassays., Results: About 80% of the LGC presented a positive M-CSF receptor staining with the immunochemical signal on cell membrane. After 72 hours culture of LGC, in absence of FSH, the baseline concentration of E(2) and P were (2,185 +/- 189) pmol/L, (3,157 +/- 401) nmol/L respectively; with the increasing dose of M-CSF from 10 to 100 ng/ml, the concentrations of E(2) increased from (2,789 +/- 365)pmol/L to (4,282 +/- 318) pmol/L, and those of P increased from (4,256 +/- 595) nmol/L to (7,789 +/- 828) nmol/L. In presence of FSH (75 IU/ml) the baseline concentrations of E(2) and P were (5,045 +/- 486) pmol/L and (8,667 +/- 923) nmol/L respectively; while the 10 - 100 ng/ml M-CSF was added the concentrations of E(2) and P were from (6,567 +/- 673) to (8,373 +/- 935) pmol/L and from (10,999 +/- 985) to (14,990 +/- 1,158) nmol/L respectively. Thus M-CSF caused a significant dose dependent increase of estradiol and progesterone production (P < 0.05), and M-CSF+ FSH further stimulated higher production of E(2) and P by LGC than M-CSF or FSH alone (P < 0.05)., Conclusions: M-CSF receptor was expressed on membrane of human LGC. M-CSF enhances estradiol and progesterone production by LGC and FSH has an additional or synergetic effect with M-CSF.