SGMS1 facilitates osteogenic differentiation of MSCs and strengthens osteogenesis-angiogenesis coupling by modulating Cer/PP2A/Akt pathway.

Mesenchymal stem cell (MSC)-mediated coupling of osteogenesis and angiogenesis is a critical phenomenon in bone formation. Herein, we investigated the role and mechanism of SGMS1 in the osteogenic differentiation of MSCs and, in combination with osteogenesis and angiogenesis, to discover new therapeutic targets for skeletal dysplasia and bone defects. SGMS1 addition accelerated MSC osteogenic differentiation, whereas SGMS1 silencing suppressed this process. Moreover, SGMS1 overexpression inhibited ceramide (Cer) and promoted sphingomyelin (SM) levels. SM treatment neutralized the suppressive effect of shSGMS1 on osteogenesis. SGMS1 restrained PP2A activity by regulating Cer/SM metabolism and thus enhanced the levels of phosphorylated Akt, Runx2, and vascular endothelial growth factor (VEGF). Furthermore, SGMS1 transcription was regulated by Runx2. SGMS1 increased MSC-mediated angiogenesis by promoting VEGF expression. SGMS1 addition promoted rat bone regeneration in vivo. In conclusion, SGMS1 induces osteogenic differentiation of MSCs and osteogenic-angiogenic coupling through the regulation of the Cer/PP2A/Akt signaling pathway.

Baicalein improves the symptoms of polycystic ovary syndrome by mitigating oxidative stress and ferroptosis in the ovary and gravid placenta.

BACKGROUND: Polycystic ovary syndrome is a metabolic and hormonal disorder that is closely linked to oxidative stress. Within individuals diagnosed with PCOS, changes occur in the ovaries, resulting in an excessive buildup of iron and peroxidation of lipids, both of which may be associated with the occurrence of ferroptosis. Baicalein, a flavonoid found in the roots of Scutellaria baicalensis and widely known as Chinese skullcap, is known for its anti-inflammatory and anti-ferroptotic properties, which protect against various diseases. Nevertheless, there has been no investigation into the impact of baicalein on polycystic ovary syndrome. PURPOSE: This study aimed to correlate ferroptosis with polycystic ovary syndrome and to assess the effects of baicalein on ovarian dysfunction and placental development in pregnant patients. STUDY DESIGN AND METHODS: Polycystic ovary syndrome was induced in a rat model through the administration of dehydroepiandrosterone, and these rats were treated with baicalein. Oxidative stress and inflammation levels were assessed in serum and ovaries, and tissue samples were collected for histological and protein analyses. Furthermore, different groups of female rats were mated with male rats to observe pregnancy outcomes and tissue samples were obtained for histological, protein, and RNA sequencing. Then, RNA sequencing of the placenta was performed to determine the key genes involved in ferroptosis negative regulation (FNR) signatures. RESULTS: Baicalein was shown to reduce ovarian oxidative stress and pathology. Baicalein also ameliorated polycystic ovary syndrome by decreasing lipid peroxidation and chronic inflammation and modulating mitochondrial functions and ferroptosis in the ovaries. Specifically, glutathione peroxidase and ferritin heavy chain 1 were considerably downregulated in polycystic ovary syndrome gravid rats compared to their expression in the control group, and most of these differences were reversed after baicalein intervention. CONCLUSIONS: Our findings, initially, indicated that baicalein could potentially enhance the prognosis of individuals suffering from polycystic ovary syndrome by reducing oxidative stress and ferroptosis, thus potentially influencing the formulation of a therapeutic approach to address this condition.

Prenatal Cases Reflect the Complexity of the COL1A1/2 Associated Osteogenesis Imperfecta.

INTRODUCTION: Osteogenesis imperfecta (OI) is a rare mendelian skeletal dysplasia with autosomal dominant or recessive inheritance pattern, and almost the most common primary osteoporosis in prenatal settings. The diversity of clinical presentation and genetic etiology in prenatal OI cases presents a challenge to counseling yet has seldom been discussed in previous studies. METHODS: Ten cases with suspected fetal OI were enrolled and submitted to a genetic detection using conventional karyotyping, chromosomal microarray analysis (CMA), and whole-exome sequencing (WES). Sanger sequencing was used as the validation method for potential diagnostic variants. In silico analysis of specific missense variants was also performed. RESULTS: The karyotyping and CMA results of these cases were normal, while WES identified OI-associated variants in the COL1A1/2 genes in all ten cases. Six of these variants were novel. Additionally, four cases here exhibited distinctive clinical and/or genetic characteristics, including the situations of intrafamilial phenotypic variability, parental mosaicism, and "dual nosogenesis" (mutations in collagen I and another gene). CONCLUSION: Our study not only expands the spectrum of COL1A1/2-related OI, but also highlights the complexity that occurs in prenatal OI and the importance of clarifying its pathogenic mechanisms.

Immune regulation in polycystic ovary syndrome.

A polycystic ovarian syndrome (PCOS) is the most common endocrine disorder affecting females. Furthermore, it is a heterogeneous disease with a variety of etiologies and outcomes. Patients frequently complain about infertility, irregular menstruation, acne, seborrheic dermatitis, hirsutism, and obesity. PCOS can be caused by hypothalamic-pituitary-ovarian axis dysfunction, heredity, or metabolic abnormalities. PCOS is characterized by chronic low-level inflammation, which includes an imbalance in pro-inflammatory factor secretion, endothelial cell dysfunction, and leukocytosis. PCOS is also distinguished by hormonal and immune dysregulation. During PCOS, immune cells and immune regulatory molecules play critical roles in maintaining metabolic homeostasis and regulating immune responses. Because of oligo/anovulation, patients with PCOS have low progesterone levels. Therefore, low progesterone levels in PCOS overstimulate the immune system, causing it to produce more estrogen, which leads to a variety of autoantibodies. This review aims to summarize the immune regulation involved in the pathogenesis of PCOS and pave the way for the development of better PCOS treatment options in the near future.

Congenital bilateral cryptorchidism in an infant conceived after maternal breast cancer treatment: A case report.

BACKGROUND: The incidence of breast cancer among women of reproductive age is increasing, as well as the desire for children at late childbearing age. Identifying factors that may be associated with fetal malformation and maternal and fetal prognosis has gained importance. We describe a 32-year-old woman with breast cancer who gave birth to a son with congenital bilateral cryptorchidism after treatment, with a literature review performed. CASE SUMMARY: A 32-year-old woman with breast cancer who had been treated by surgery and radiotherapy experienced recurrence and underwent a second surgery, adjuvant chemotherapy, and targeted therapy. Her tumor cells were negative for estrogen receptor (ER) α, progesterone receptor (PR), and p53; positive for ERß, human epidermal growth factor receptor-2 (HER2), epidermal growth factor receptor (EGFR), and Ki67. She had pathogenic BRCA gene mutations. She became pregnant within 2 years and delivered a boy with congenital bilateral cryptorchidism. The boy underwent bilateral orchidopexy. As of this writing, the woman and her son are both healthy. CONCLUSION: HER2 overexpression, positivity for EGFR, Ki67, and ER, and PR negativity are associated with a poor prognosis in breast cancer. While no link has been established statistically between treatment for breast cancer and cryptorchidism in a subsequent pregnancy, this case suggests the possibility that ERß and gene mutations may be contributing factors.

Prediction of postpartum hemorrhage in pregnant women with immune thrombocytopenia: Development and validation of the MONITOR model in a nationwide multicenter study.

Globally, postpartum hemorrhage (PPH) is the leading cause of maternal death. Women with immune thrombocytopenia (ITP) are at increased risk of developing PPH. Early identification of PPH helps to prevent adverse outcomes, but is underused because clinicians do not have a tool to predict PPH for women with ITP. We therefore conducted a nationwide multicenter retrospective study to develop and validate a prediction model of PPH in patients with ITP. We included 432 pregnant women (677 pregnancies) with primary ITP from 18 academic tertiary centers in China from January 2008 to August 2018. A total of 157 (23.2%) pregnancies experienced PPH. The derivation cohort included 450 pregnancies. For the validation cohort, we included 117 pregnancies in the temporal validation cohort and 110 pregnancies in the geographical validation cohort. We assessed 25 clinical parameters as candidate predictors and used multivariable logistic regression to develop our prediction model. The final model included seven variables and was named MONITOR (maternal complication, WHO bleeding score, antepartum platelet transfusion, placental abnormalities, platelet count, previous uterine surgery, and primiparity). We established an easy-to-use risk heatmap and risk score of PPH based on the seven risk factors. We externally validated this model using both a temporal validation cohort and a geographical validation cohort. The MONITOR model had an AUC of 0.868 (95% CI 0.828-0.909) in internal validation, 0.869 (95% CI 0.802-0.937) in the temporal validation, and 0.811 (95% CI 0.713-0.908) in the geographical validation. Calibration plots demonstrated good agreement between MONITOR-predicted probability and actual observation in both internal validation and external validation. Therefore, we developed and validated a very accurate prediction model for PPH. We hope that the model will contribute to more precise clinical care, decreased adverse outcomes, and better health care resource allocation.

Killing cervical cancer cells by specific chimeric antigen receptor-modified T cells.

The aim is to investigate the in vivo and in vitro killing effect of mesothelin chimeric antigen receptor T cells (MESO-CAR-T) in cervical squamous cell carcinoma. MESO-CAR-T cells were successfully constructed. In vitro verification of the killing effect of MESO-CAR-T cells was evaluated in the presence of SiHa cells by the lactate dehydrogenase release assay and cytokine release assay. The in vivo experiments were performed in immunodeficient NCG mice. After successful tumor formation with the subcutaneous implantation of SiHa cervical cancer cells, the injections of MESO-CAR-T cells into the tumors at different doses and frequencies were performed. Subsequently, the growth rate and size of the tumors in NCG mice were observed. A 17-fold increase in the number of MESO-CAR-T cells and a 16-fold increase in the number of Con-CAR-T cells were observed. The result of marker detection in the prepared MESO-CAR-T cells showed that CD3+ T lymphocytes accounted for 97.0 % of all cells, indicating successful preparation of MESO-CAR-T cells. Expression of the membrane protein MESO was detected in 12.8 % of SiHa cells. When the ratio of MESO-CAR-T cells to SiHa cells was 20:1, the lysis of target cells was most significant and was observed in 22 % of the cells. In the presence of SiHa cells, the secretion of IL-4、IL-2、IL-5、TNF-α and IFN-γ in MESO-CAR-T cells was higher than that in the control group. The effect of two consecutive intratumoral injections of MESO-CAR-T cells was more obvious than that of one injection. The pharmacological effect of the injection was observed within 1 week. Our finding identified the certain in vivo and in vitro killing activity of MESO-CAR-T cells.

Use of mesothelin as a tumor-associated antigen in cervical squamous cell carcinoma.

AIM: To explore the feasibility of using mesothelin (MESO) as a tumor-associated antigen (TAA) in cervical squamous cell carcinoma and its function in the development of cervical cancer. METHODS: We collected eight cervical tissue samples of squamous cell carcinoma as the test group and eight samples of cervicitis as the control group from patients who underwent a hysterectomy because of a diagnosis of myoma. Then we used western blotting to screen for a potential TAA in cervical squamous cell carcinoma samples. In addition, Lentivirus-mediated RNAi was used to downregulate the expression of the MESO gene (MSLN) in SiHa cells. Fluorescence-activated cell sorting (FACS), 3­(4,5)-dimethylthiahiazo (-z­y1)­3,5­di­phenytetrazoliumromide (MTT), and wound healing were used to examine cell apoptosis, cell proliferation, and cell migration respectively. RESULTS: Results of the western blotting showed that the MESO protein expressed highly in the cervical squamous cell carcinoma and paracancerous tissues in contrast to the cervicitis tissue (p = 0.242). We used quantitative PCR to verify that the expression of MSLN was 21.6% in the cells undergoing knockdown compared to that in the control cells, and thus, decided to continue with further experiments. We confirmed by FACS that the apoptosis rate in the SiHa cell group undergoing MSLN knockdown (KD group) was significantly higher than that in negative control (NC) group of SiHa cells (p = 0.014). The proliferation of cells was examined by MTT and the difference between the KD and NC groups was found to be statistically significant (p = 0.002). However, since the difference was <20% we did not consider it to be clinically significant. Cell migration ability was verified by wound healing test and found to be 43% in the KD group and 38% in the NC group after 48 h, but the difference was not statistically significant (p > 0.05). CONCLUSIONS: MESO might be used as a TAA for diagnosing cervical squamous cell carcinoma. When MSLN was knocked down in SiHa cells, cell apoptosis increased, but no significant effects were observed on cell proliferation and migration. Thus, our study shows that MSLN plays a role in the apoptosis of cervical squamous cell carcinoma cells, and since this might affect tumor progression, further research is warranted to understand how MSLN plays this role.

A clinical analysis of small-cell neuroendocrine carcinoma of the gynecologic tract: report of 20 cases.

OBJECTIVE: The aim of this retrospective observational study was to analyze the clinical and pathological characteristics of small-cell neuroendocrine carcinoma of the gynecologic tract (SCNCGT). METHODS: Twenty patients with SCNCGT were enrolled and their clinic-pathological features were analyzed. All patients were treated at the Beijing Obstetrics and Gynecology Hospital, Capital Medical University, China, and were followed up until December 31, 2017. RESULTS: (1) Patient characteristics: The incidence of SCNCGT was 0.3% (20/6578) of gynecologic cancer in our hospital from January 1, 2007, to December 31, 2017. The average age of the patients was 42.0 ± 11.8 (23-63 years). Out of 20 patients enrolled, seven (35.0%) had lymph node metastasis. Out of 17 patients treated with complete surgery, 14 (82.4%) had lymph-vascular space invasion. (2) Treatment: Eleven out of the 14 patients with small-cell neuroendocrine carcinoma of the cervix (SCNCC) were treated with radical surgery; all the 11 patients received chemotherapy and radiotherapy postoperatively. The remaining three patients received comprehensive chemotherapy and/or radiotherapy instead of radical surgery. The six patients who had one or the other type of SCNCGT (involving the ovary, endometrium, or vagina) were all treated with comprehensive surgery. (3) Prognosis: The follow-up time for the study ranged from 8 to 87 months. Three (15.0%) of the 20 patients were diagnosed with distant metastasis at the beginning of the study. Eight (40.0%) patients died as of December 31, 2017, while the other 12 patients were in follow-up. The average survival time was 43.6 months (16-77 months). CONCLUSION: SCNCGT is a highly malignant tumor characterized by rare morbidity, a propensity for metastasis, and poor prognosis. Comprehensive treatment may be a good approach to prolong survival in some patients.

Effect of Dandelion Extracts on the Proliferation of Ovarian Granulosa Cells and Expression of Hormone Receptors.

BACKGROUND: In the current society, infertility related to age has become a social problem. The in vitro fertilization (IVF) success rate in women with poor ovarian response (POR) is very low. Dandelion extract T-1 (DE-T1) is an effective component of the extract from the leaves and stems of Taraxacum officinale, which is one of the medicines used in some patients with POR, but its molecular mechanism remains unclear. METHODS: Following IVF, ovarian granulosa cells (GCs) of sixty patients were extracted and divided into normal ovarian response (NOR) and POR groups. GCs were cultured in a dose-dependent and time-dependent manner with DE-T1, proliferation of GCs was determined by Cell Counting Kit-8 assay, and mRNA levels of insulin-like growth factor 1 receptor (IGF-1R), luteotropic hormone receptor (LHR), follicle-stimulating hormone receptor (FSHR), LHR, and CYP19A1 (aromatase) were determined by quantitative polymerase chain reaction. Progesterone and estradiol (E2) concentrations were determined by enzyme-linked immunosorbent assay. RESULTS: The cell viability gradually increased with the progressive increase in the DE-T1 concentration. Compared with the control group (without DE-T1), the mRNA expressions of FSHR, LHR, IGF-1R, and CYP19A1 were upregulated after the addition of DE-T1, especially in the 2.5% DE-T1 group (P < 0.01). The expression of IGF-1R was upregulated approximately 25 times (24.97 ± 4.02 times) in the POR group with 2.5% DE-T1. E2 and progesterone levels increased with the increasing DE-T1 concentration. There were highly significant differences in the E2 and progesterone secretion between the NOR and POR groups (P < 0.01). CONCLUSION: DE-T1 may promote steroid hormone synthesis by promoting GC proliferation and upregulating GC receptor expression, thereby improving ovarian endocrine function.

Analysis of short-term efficacy as defined by RECIST and pathological response of neoadjuvant chemotherapy comprised paclitaxel and cisplatin followed by radical surgery in patients with locally advanced cervical cancer: A prospective observational study.

The purpose of this study is to investigate short-term efficacy as defined by the Response Evaluation Criteria in Solid Tumors (RECIST) and pathological response of neoadjuvant chemotherapy (NACT) comprised of paclitaxel and cisplatin (TP) followed by radical surgery in patients with locally advanced cervical cancer (LACC).This is a prospective study involving 61 women with histologically confirmed LACC referred for NACT following radical surgery at Beijing Obstetrics and Gynecology Hospital between April 2013 and January 2015.The efficacy of NACT was evaluated by the RECIST. The total short-term efficacy of NACT was 91.8% (complete remission and partial remission). The cervical invasion ≤1/2 was 82.4% in the complete remission (CR) group, 46.2% in the partial remission (PR) group, and 20% in the stable disease (SD) group. The difference between groups was statistically significant (P = .012). The slides of all surgical specimens were reviewed and classified according to the Tumor Regression Grade (TRG). The good response was defined by good short-term efficacy (RECIST) and the difference between groups was statistically significant (P = .042). The route of administration of NACT is a factor predicting response to NACT. A significant higher response rate (P = .011) and lower chemotherapy-related adverse events (P < .05) were observed in the artery intervention (AI) group compared to those received NACT via intravenous (IV) route. All patients were followed-up to the last day of 2015 with the median follow-up time of 21.5 months for NACT. For the 61 patients referred for NACT in LACC, 2 patients had relapsed and 1 patient died from the disease.The study showed that the NACT comprised TP for LACC treatment had a significant local effect. It could reduce tumor myometrial invasion and regress tumor. The route of administrating NACT is a predicting factor to the NACT response; 2 cycles of NACT of AI treatment to LACC patients would obtain a desired response with low chemotherapy adverse events.

Clinical analysis of cervical intraepithelial neoplasia with vaginal intraepithelial neoplasia.

The purpose of this prospective cohort study is to evaluate the importance of screening and its diagnostic accuracy compared with the pathological diagnosis of cervical intraepithelial neoplasia (CIN) with vaginal intraepithelial neoplasia (VAIN).The prospective study enrolled 419 patients (pts) and was conducted between February 1, 2015 and January 31, 2016 at Beijing Obstetrics and Gynecology Hospital, Capital Medical University.All enrolled pts underwent multipoint biopsy of cervix and vaginal wall directed by colposcopy. All samples of biopsy underwent pathological examination. Among them, 201 pts (48.0%) were diagnosed with CIN, 218 pts (52.0%) were diagnosed with cervicitis, and 51 pts (12.2%) were diagnosed with VAIN. It was found that the incidence of CIN in pts was 4 times higher than that of VAIN. In all 419 patients enrolled, 218 pts had cervicitis with 13 pts (6.0%) of VAIN. There were 201 pts of CIN with 38 pts (18.9%) of VAIN: including 53 pts of CIN3 with 12 pts (22.6%) of VAIN; 49 pts of CIN2 with 9 pts of VAIN (18.4%), and 99 pts of CIN1 with 17 pts of VAIN (17.2%). The incidence of CIN with VAIN (18.9%) was significantly higher than cervicitis with VAIN (6.0%) (χ = 16.39, P = .00). Our results showed that there was a significant consistency between cervical lesions and vaginal lesions (χ = 135.91, P = .00), which indicated that the increase of CIN grades may be related to an increase of the VAIN grades. Our results also showed the significant (p < .05) increase of CIN and VAIN with age (<40 years Kappa = 0.04; 40-50 years Kappa = 0.11; >50 years Kappa = 0.28).This study showed that cytological test can be used as a routine screening method for cervical lesions and vaginal diseases. If the cytology result shows abnormality, and pathological examination confirms that there is no obvious abnormal cervical disease, colposcopy directed vaginal multipoint biopsy should be conducted to exclude vaginal disease. All patients of CIN should routinely undergo vaginal multipoint biopsy (1/3 upper vagina), especially in patients with high-grade CIN and age older than 50 years.

Hydrogen-rich Water Exerting a Protective Effect on Ovarian Reserve Function in a Mouse Model of Immune Premature Ovarian Failure Induced by Zona Pellucida 3.

BACKGROUND: Premature ovarian failure (POF) is a disease that affects female fertility but has few effective treatments. Ovarian reserve function plays an important role in female fertility. Recent studies have reported that hydrogen can protect male fertility. Therefore, we explored the potential protective effect of hydrogen-rich water on ovarian reserve function through a mouse immune POF model. METHODS: To set up immune POF model, fifty female BALB/c mice were randomly divided into four groups: Control (mice consumed normal water, n = 10), hydrogen (mice consumed hydrogen-rich water, n = 10), model (mice were immunized with zona pellucida glycoprotein 3 [ZP3] and consumed normal water, n = 15), and model-hydrogen (mice were immunized with ZP3 and consumed hydrogen-rich water, n = 15) groups. After 5 weeks, mice were sacrificed. Serum anti-Müllerian hormone (AMH) levels, granulosa cell (GC) apoptotic index (AI), B-cell leukemia/lymphoma 2 (Bcl-2), and BCL2-associated X protein (Bax) expression were examined. Analyses were performed using SPSS 17.0 (SPSS Inc., Chicago, IL, USA) software. RESULTS: Immune POF model, model group exhibited markedly reduced serum AMH levels compared with those of the control group (5.41 ± 0.91 ng/ml vs. 16.23 ± 1.97 ng/ml, P = 0.033) and the hydrogen group (19.65 ± 7.82 ng/ml, P = 0.006). The model-hydrogen group displayed significantly higher AMH concentrations compared with that of the model group (15.03 ± 2.75 ng/ml vs. 5.41 ± 0.91 ng/ml, P = 0.021). The GC AI was significantly higher in the model group (21.30 ± 1.74%) than those in the control (7.06 ± 0.27%), hydrogen (5.17 ± 0.41%), and model-hydrogen groups (11.24 ± 0.58%) (all P < 0.001). The GC AI was significantly higher in the model-hydrogen group compared with that of the hydrogen group (11.24 ± 0.58% vs. 5.17 ± 0.41%, P = 0.021). Compared with those of the model group, ovarian tissue Bcl-2 levels increased (2.18 ± 0.30 vs. 3.01 ± 0.33, P = 0.045) and the Bax/Bcl-2 ratio decreased in the model-hydrogen group. CONCLUSIONS: Hydrogen-rich water may improve serum AMH levels and reduce ovarian GC apoptosis in a mouse immune POF model induced by ZP3.

[A clinical study on the diagnostic criteria of multiple organ dysfunction syndrome].

OBJECTIVE: To develop diagnostic criteria of multiple organ dysfunction syndrome (MODS) by a prospective and multi-center clinical investigation. METHODS: The data of 1087 MODS cases obtained from ICU of 37 hospitals from March 2002 to January 2005 in 11 provinces in China were analyzed in order to derive the diagnostic criteria of MODS. RESULTS: This MODS diagnostic criteria involved 7 organs. To diagnose MODS, the original cause of MODS should be identified, then there should be two or more organs showing signs of dysfunction. The criteria for organ dysfunction were as follows. (1) Cardiovascular system: SBP < 90 mm Hg (1 mm Hg = 0.133 kPa), MAP < 70 mm Hg, signs of shock, ventricular tachycardia, ventricular fibrillation, or myocardial infarction; (2) Respiratory system: oxygenation index < 300 mm Hg; (3) Nervous system: indifference, restlessness, lethargy, light coma, or deep coma, Glasgow score < or = 14; (4) Blood system: PLT < 100 x 10(9)/L; CT, APTT, and PT prolonged or shortened; positive plasma protamine paracoagulation; (5) Liver: TBIL > 20. 5 micromol/L, ALB < 28 g/L; (6) Kidney: Cr > 123.8 micromol/L, urinary volume < 500 ml/24 h; (7) Gastro-intestine: bowel sounds decreased or disappeared; retention in the stomach, or positive occult blood feces with dark stools or haematemesis; intraabdominal pressure (intravesical pressure) > or = 11 cm H2O (1 cm H2O = 0.098 kPa). Any organ function met with one of the above conditions was considered to have dysfunction. CONCLUSIONS: This diagnostic criterion of multiple organ dysfunction syndrome has been developed by this research, but it needs to accumulate experience by clinical practice and to revise the diagnosis criteria.

[Effects of cervical chordotomy on systemic inflammatory response and the outcome in rats with endotoxic shock induced by lipopolysaccharide].

OBJECTIVE: To investigate the effects of cervical chordotomy on systemic inflammatory response and the outcome in rats with endotoxemia induced by lipopolysaccharide (LPS). METHODS: Ninety-two Sprague Dawley (SD) rats were randomly divided into three groups: normal control group (group I, n=8) , endotoxemia group (group II, n=42) and endotoxemia with cervical chordotomy (group III, n=42). Endotoxemia was induced by intra-peritoneal injection of LPS 10 mg/kg. In group III, "cervical chordotomy" was attained by transection of spinal cord at C7 immediately before intra-peritoneal LPS administration. Ten rats of group II and III each were observed for 48-hour survival. The other rats were further divided into four subgroups of 8 animals each, according to the time when the animals were sacrificed . The animals were sacrificed at 3, 6, 12, and 48 hours after intra-peritoneal LPS injection. Heart blood samples were obtained for determination of plasma concentration of norepinephrine [NE, by high performance liquid chromatography (HPLC)] and plasma concentration of interleukin-10 (IL-10) and IL-6 [by enzyme linked immunosorbent assay (ELISA)]. RESULTS: Plasma NE concentration were significantly increased after intra-peritoneal LPS injection in group II and III as compared with group I and were significantly lower in group III than in group II starting from 6 hours after LPS (all P<0.05). Plasma IL-10 concentration was significantly lower at 3 hours and 6 hours while plasma IL-6 concentration was significantly higher after LPS challenge in group II than in group I at all time points (all P<0.05). High transection of spinal cord significantly elevated plasma IL-10 level at 12 hours and 48 hours, lowered IL-6 release at 3, 6, and 12 hours (all P<0.05), and improved 48-hour survival (20% vs. 70%) in group III as compared with group II. CONCLUSION: Transection of spinal cord at C7 level can ameliorate the systemic inflammatory response induced by endotoxemia thus improving the outcome through elevation in IL-10 level, decreases in IL-6 release, and improves 48-hour survival. This might be attributable to loss of sympathetic nerve function.

[Liver injury and intervention of compound 912 liquid on it in rats with endotoxemia].

OBJECTIVE: To investigate the liver injury in model rats with endotoxemia and to observe the protective effect of Compound 912 Liquid on it. METHODS: Rats were randomly divided into three groups, the endotoxemia model group (EMG, injected by lipoplysaccharides (LPS) peritoneally), the intervention group (IG, treated with Compound 912 Liquid via gastrogavage 1 h before model establishing) and the normal control group (NCG). Blood samples of rats were taken at the time points of the 2nd, 4th, 8th, 12th, 48th, 72nd hour and the 7th day after modeling for measuring liver function, levels of plasmatic endotoxin, tumor necrosis factor alpha (TNF-alpha), interleukin-10 (IL-10). The pathological change of liver was observed using light microscope and electro-transmission microscope. RESULTS: The peak concentration of endotoxin detected at 2 hour after modeling in the IG was significantly lower than that in the EMG (0.358 +/- 0.056 vs 0.685 +/- 0.030), but insignificant difference (P > 0.05) was shown between them in TNF-alpha level. The level of IL-10 continuously rose in IG after treatment, it was still higher than normal level until day 7 (49.096 +/- 4.076 vs 43.454 +/- 5.928, P < 0.05). CONCLUSION: LPS can induce the increase of serum inflammatory cytokines and anti-inflammatory cytokines in rats to injure liver. Therefore, the inflammatory reaction indicated by LPS may be one of the mechanisms for liver injury. Preventive medication with Compound 912 Liquid showed a significant liver protective effect.

[Effect of treatment with Xuebijing injection on serum inflammatory mediators and Th1/2 of spleen in rats with sepsis].

OBJECTIVE: To study the effect of treatment with Xuebijing injection on pro- or anti-inflammatory response and pathologic changes in immune organs in rats with sepsis. METHODS: Sepsis was reproduced in rats by cecal ligation and puncture. Rats were divided into four groups i.e. sham operation, sepsis, sepsis with levofloxacin treatment, and Xuebijing treatment groups. Blood samples were collected at 3, 24 and 72 hours after model was reproduced. Enzyme linked immunoadsorbent assay (ELISA) was used to determine the levels of serum tumor necrosis factor-alpha (TNF-alpha) and interleukin-10 (IL-10), expression level of spleen Th1/Th2 was assessed by FACS flow cytometer, and the pathological changes in immunological organs were examined. RESULTS: The results showed that the levels of Th1/Th2 and TNF-alpha, IL-10 were elevated in early period of sepsis, but lowered in late period of sepsis. Xuebijing could decrease the level of Th1/Th2, which showed an increase at 72 hours. Xuebijing could lower the levels of TNF-alpha and IL-10, rendering a balance of pro- and anti-inflammatory response. CONCLUSION: There is a dissonance in immunological function in sepsis, showing a depression in specific immunological function, but an exaggeration in non-specific immunological function. Xuebijing can obviously ameliorate the immunological disturbance in sepsis of rat.

[Protective effects of dexamethasone on myocardium in rats with sepsis].

OBJECTIVE: To investigate the changes in heart function and myocardial mechanics in murine sepsis model, and the mechanism of the protective effect of dexamethasone on heart. METHODS: Wistar rats were randomly divided into control group, lipopolysaccharide (LPS) group (4 mg/kg LPS intravenously), LPS+dexamethasone group (4 mg/kg LPS+2 mg/kg dexamethasone intravenously), with 32 rats in each group. A catheter was passed through right common carotid artery to the left cardiac ventricle. Function of the left ventricle was monitored, and blood was drawn at 0, 2, 4, 6 hours to detect concentrations of tumor necrosis factor-alpha (TNF-alpha), troponin T (TnT), with 8 rats for each time point. RESULTS: In sepsis rats, TnT increased significantly and could be lowered by dexamethasone [at 6 hours after the treatment (1.76+/-0.57) microg/L vs. (0.70+/-0.36) microg/L, P<0.01]. There were changes in left ventricular peak systolic pressure (LVPP) and maximum rate of intraventricular pressure rise/down (+/-dp/dt max) to certain extent, and increase in left ventricular end-diastolic pressure (LVEDP), but these changes could be ameliorated by using dexamethasone. TNF-alpha increased significantly in sepsis rats, but dexamethasone could lower its level [at 2 hours after the treatment (11.22+/-2.38) pmol/L vs. (7.62+/-3.21) pmol/L, P<0.01]. CONCLUSION: Myocardium is remarkably damaged in rats with sepsis. TNF-alpha could be regarded as one of the factors which could produce injury to myocardium. Dexamethasone could alleviate cardiac damage produced by endotoxin in sepsis model.

[Study on changes in circulatory renin-angiotensin system in rats with endotoxemia].

OBJECTIVE: To investigate the changes in circulatory renin-angiotensin system (RAS) in rats with endotoxemia and explore its mechanism. METHODS: Ninety Wistar rats were randomly divided into normal control group (n=10) and endotoxemia groups (n=80). The endotoxemia group was further divided into subgroups of 2, 4, 8, 12, 24, 48, 72 hours and 7 days after injection of lipopolysaccharide (LPS) into the abdominal cavity (each n=10). The rats were sacrificed at different time points after LPS (10 mg/kg) challenge. Contents of plasma LPS, plasma renin activity (PRA), plasma angiotensin II (A II), angiotensin-converting enzyme (ACE), tumor necrosis factor-alpha (TNF-alpha), and interleukin-10 (IL-10) were determined. RESULTS: (1) Plasma LPS, TNF-alpha and IL-10 reached highest levels 2 hours after LPS injection, and they returned to normal levels 7 days, 8 hours and 8 hours after LPS challenge respectively. (2) The levels of plasma PRA, A II and ACE reached highest levels 2, 8 and 8 hours after LPS challenge respectively, and they returned to normal levels 12, 24 and 72 hours after LPS injection, respectively. CONCLUSION: During endotoxemia, there is a series of inflammatory reactions and anti-inflammatory reactions, and a large number of cytokines and inflammatory mediators are released. At the same time, the circulating RAS is activated, which causes constriction of blood vessel and dysfunction of the microcirculation aggravating hypoxia of cells. The study of the changes in RAS is significant to interpreting the pathogenetic mechanism of endotoxemia in rats.