This study investigated the growth-regulating effects of progesterone (Prog) on nPR-negative malignant melanoma cells and the possible mechanisms. A375 and A875 cells were cultured and treated with Prog of different concentrations. For signal transduction pathway studies, the cells were pretreated with Prog receptor antagonist (RU486, 1x10(-7) mol/L) or MAPK inhibitor (U0126, 5x10(-6) mol/L) for 1 h and then co-incubated with prog (10(-9) mol/L) for another 24 h. Indirect immunofluorescence assay, MTT, flow cytometry and Western blotting were used for assessing the nPR expression, cell growth, cell apoptosis and ERK1/2 Phosphorylation, respectively. Our results showed that lower progesterone concentration promoted the proliferation of both A375 and A875 cells, but this growth-stimulatory effect decreased at progesterone concentration of 1x10(-7) mol/L or higher. The response could be abolished by MAPK inhibitor U0126, but could not be blocked by progesterone antagonist RU486. Flow cytometry exhibited that high concentration ([Symbol: see text]1 x 10(-7) mol/L) progesterone increased the apoptosis of the two cells in a dose-dependent manner. The level of ERK1/2 phosphorylation was increased by a lower progesterone concentration, but reduced by a higher concentration (1x10(-6) mol/L). These results suggest progesterone exerts growth-regulating effects on nPR-negative tumor cells through a non-genomic mechanism.

AIM To study the influence of Huangqi Powder (Puerariae lobatae Radix,Astragali Radix,Mori Cortex) on femur and tibia in rats with glucocorticoid-induced osteoporosis (GIOP).METHODS Thirty one-month-old male SD rats were divided into three groups by randomized block design,which were control group (normal saline),prednisone group (3.5 mg/kg prednisone acetate),Huangqi Powder group (clinical adult dosage).Experimental test was maintained for one hundred and twenty days.Double fluorochrome labeling with calcein was performed before necropsy;bone resorption and bone formation index were determined by using blood serum;the femur structural and material mechanics were analyzed by using bone biomechanics method.The cancellous bone of proximal tibial and the cortical bone of tibial shaft static parameter and dynamic parameter were analyzed by bone histomorphometry method.RESULTS Huangqi Powder had no obvious improving effects on bone biomechanics and bone static parameter,but could increas the area,width and number of the proximal tibia bone trabecular.Mark perimeter percentage and mineralization rate of deposition could be increased.CONCLUSION Huangqi Powder can significantly increase proximal tibia cancellous bone mass and enhance bone trabecula structure of glucocorticoid-induced osteoporosis rats.

Objective To evaluate the suitability of preoperative magnetic resonance imaging (MRI) positivity as a pre?dictor of biochemical recurrence after radical prostatectomy (RP). Methods Data of 115 diagnosed prostate cancer patients with RP from 2006 to 2013 were collected retrospectively. According to MRI results, patients were divided into MRI positive group (n=87) and MRI negative group (n=28). The preoperative clinical data were compared between two groups. Biochemi?cal recurrence was defined as increase or persistence of serum prostate specific antigen (PSA) levels>0.2μg/L after surgery. Cox proportional hazard regression model was used to analyze the risk factors associated with biochemical recurrence. Re?sults The survival time was shorter in MRI positive group than that of MRI negative group. The preoperative PSA level was higher in MRI positive group than that of MRI negative group (P<0.05). There was no significant difference in the age of pa?tients between two groups. In MRI positive group, the proportion of preoperative PSA>20μg/L , the proportion of patients at clinical stage T3, the proportion of patients with pathological Gleason score≥8, the proportion of patients with seminal vesi?cle invasion (SVI) and the proportion of patients with endocrine therapy were higher than those of negative group ( P<0.05). The univariate analysis showed that preoperative MRI positivity, PSA level, biopsy Gleason score, clinical stage, pathological Gleason score, SVI level and lymph node invasion (LNM) were associated with biochemical recurrence (P<0.05).However, the multivariate analysis showed that only preoperative MRI positivity, biopsy Gleason score and clinical stage were indepen?dent prognostic factors (P<0.05). Conclusion Preoperative MRI positivity can predict biochemical recurrence after RP successfully, and improve the prognosis of patients.

Objective To observe the change of peripheral blood CD34+ cell level in patients with acute cerebral infarction, and explore its relationships with cerebrovascular risk factors,neurological function and carotid artery intima-media thickness (IMT). Methods The 45 patients with acute cerebral infarction (onset within 72 h) (infarction group) and 27 patients with cerebr ovascular risk factors but without cerebral infarction (high-risk group) were chosen for the study. The cerebrovascular disease risk factors including history of alcohol abuse, smoking, coronary heart disease, hypertension, diabetes, abnormal levels of serum triglycerides, total cholesterol,low-density lipoprotein cholesterol (LDL-C), and high density lipoprotein cholesterol (HDL-C) were recorded in all subjects. The peripheral blood CD34+ cell levels were measured by flow cytometry.The correlations of peripheral blood CD34+ cell level with cerebrovascular disease risk factors were analyzed. The neurological function and carotid artery IMT were recorded in infarction group, and the correlations of peripheral blood CD34+ cell level with neurological function and carotid artery IMT were analyzed. Results (1) The peripheral blood CD34+ cell level was significantly negatively correlated with coronary heart disease, hypertension, diabetes and LDL-C level (r =- 0. 749,-0. 717, - 0. 688, - 0. 764, all P＜0. 01) ; (2) Multiple linear regression analysis showed that peripheral blood CD34+ cell level was an independent relative factor of acute cerebral infarction (P＜0.05); (3) The peripheral blood CD34+ cell level was lower in infarction group than in high-risk group, and was significantly negatively correlated with neurological deficit score (r=-0. 721, P＜0.01) and carotid artery IMT (r= -0. 695, P＜0. 01). Conclusions Peripheral blood CD34+ cell level could be an independent relative factor of acute cerebral infarction; The peripheral blood CD34+ cell level is significantly negatively correlated with neurological function and carotid artery IMT in patients with acute cerebral infarction; And it can be used as cytological marker which reflect early vascular endothelial function in patients with ischemic stroke.

Objective:To evaluate the clinical efficacy of acupuncture in the treatment of anxiety state in patients with primary insomnia(PI). Methods:Randomized controlled trials of acupuncture treatment for PI patients with an anxiety state in Web of Science,Cochrane Library,PubMed,Excerpta Medica Database(EMBASE),China National Knowledge Infrastructure(CNKI),Wanfang Data Knowledge Service Platform(Wanfang),and Chongqing VIP Database(VIP)were retrieved by computer.The retrieval time was from each database's inception to December 30,2022.Data extraction and evaluation were performed for the included studies.The Cochrane risk of bias assessment tool was used to assess the risk of bias in each article.Meta-analysis of valid data was performed using the RevMan 5.4 software.If the outcome indicator was a categorical variable,relative risk(RR)was used as the effect size.If it was a continuous variable,mean difference(MD)was used to calculate the effect size.Each effect size was expressed as a 95%confidence interval(CI).P<0.05 was considered to indicate a statistically significant difference. Results:A total of 18 studies were included,comprising a total of 1198 patients.The findings of the meta-analysis showed that electroacupuncture had a significant advantage in improving the Hamilton anxiety scale(HAMA)score than benzodiazepines[MD=-1.61,95%CI(-2.17,-1.06),P<0.001].Acupuncture was superior to sham acupuncture[MD=-14.90,95%CI(-20.39,-9.41),P<0.001]and benzodiazepines[MD=-3.39,95%CI(-4.67,-2.12),P<0.001]in reducing the self-rating anxiety scale(SAS)score.Acupuncture was superior to sham acupuncture in reducing the insomnia severity index(ISI)score[MD=-5.61,95%CI(-6.63,-4.89),P<0.001].Acupuncture was superior to benzodiazepines[MD=0.84,95%CI(-1.42,-0.25),P=0.005]and sham acupuncture[MD=-8.39,95%CI(-8.39,-7.86),P<0.001]in improving the Pittsburgh sleep quality index(PSQI)score.Acupuncture had a better effective rate than benzodiazepines[RR=1.16,95%CI(1.08,1.25),P<0.001]and sham acupuncture[RR=8.94,95%CI(4.63,17.25),P<0.001]in treating PI. Conclusion:Acupuncture or electroacupuncture has certain therapeutic advantages over benzodiazepines and sham acupuncture in the treatment of anxiety in PI patients.However,more high-quality randomized controlled trials are needed for further verification.

Human leukocyte antigen G (HLA-G) is one of the molecules implicated in immunotolerance. To investigate the role of HLA-G in primary cutaneous malignant melanoma (CMM), a series of 47 skin melanocytic lesions were immunohistochemicaily evaluated. The correlation between HLA-G expression and CMM clinicohistopahtologicai data and Bcl-2 expression was also analyzed.HLA-G expression was detected in a variety of cell types. No significant difference in HLA-G expression was observed between malignant and non-malignant melanocytic lesions. HLA-G expres- sion was significantly correlated with the inflammatory infiltration and Bel-2 expression, whereas no significant correlation with ulceration, tumor thickness, clinical stage, histopathologicai subtypes were observed. HLA-G expression may be the result of host immune reaction in tumor microenvi- ronment rather than a malignant feature of CMM.

Objective To investigate the predictive value of serum human chorionic gonadotropin(HCG),D-dimer and granulocyte colony-stimulating factor(G-CSF)levels for pregnancy outcome in patients with re-current implantation failure(RIF).Methods A total of 80 RIF patients who underwent in vitro fertilization-embryo transfer in this hospital from May 2019 to May 2022 were selected as the research objects.According to the occurrence of adverse pregnancy outcomes,they were divided into a good pregnancy outcome group(48 cases)and a poor pregnancy outcome group(32 cases).The levels of serum HCG,D-dimer and G-CSF in RIF patients at different pregnancy stages(the day of pregnancy test,the first trimester,the second trimester and the third trimester)were detected.Spearman correlation was used to analyze the correlation between serum levels of HCG,D-dimer,and G-CSF and pregnancy outcomes.Multivariate Logistic regression was used to an-alyze the influencing factors of pregnancy outcome in patients with RIF.Receiver operating characteristic(ROC)curve was used to analyze the predictive value of serum HCG,D-dimer and G-CSF levels for pregnancy outcome in patients with RIF.Results Adverse pregnancy occurred in 32 cases of 80 patients with RIF,and preterm birth and intrauterine distress were the most common adverse pregnancy outcomes.The RIF patients with different stages of pregnancy serum HCG levels gradually decreased,serum D-dimer,G-CSF level in-creased(P<0.05).At different stages of pregnancy,the level of serum HCG in the poor pregnancy outcome group was lower than that in the good pregnancy outcome group(P<0.05),and the levels of serum D-dimer and G-CSF were higher than those in the good pregnancy outcome group(P<0.05).The level of serum HCG was negatively correlated with the adverse pregnancy outcome(r=-0.506,P<0.05),and the levels of ser-um D-dimer and G-CSF were positively correlated with the adverse pregnancy outcome(r=0.498,0.542,P<0.05).Multivariate Logistic regression analysis showed that HCG,D-dimer and G-CSF levels were the influ-encing factors of poor pregnancy outcome in patients with RIF(P<0.05).Serum HCG,D-dimer,G-CSF pa-tients with single and combined forecast RIF area under the curve of the pregnancy outcomes were 0.877,0.947,0.911 and 0.971,the sensitivity were 88.10%,88.10%,90.48%and 95.24%respectively,the specific-ity were 81.58%,97.37%,76.32%and 89.47%,respectively.The combined detection of the three markers could improve the prediction efficiency(P<0.05).Conclusion The combined detection of serum HCG,D-di-mer and G-CSF levels can effectively predict the pregnancy outcome of patients with RIF.

Objective:To observe multiple locus variable-number tandem repeat analysis (MLVA) typing of Brucella isolated from Himalayan marmot in Qinghai-Tibet Plateau of Qinghai Province, and to explore the relationship between the strains and strains previous isolated from Qinghai Province. Methods:Blood samples of Himalayan marmot were collected in Qinghai-Tibet Plateau of Qinghai Province from March 2019 to October 2020. Pathogens were isolated and cultured from Brucella antibody positive samples identified by using the rose bengal test (RBT). Conventional biological methods and molecular biological methods (BCSP31-PCR and AMOS-PCR) were used for strain identification. At the same time, MLVA method was used to genotype the isolated strains, and cluster analysis was used to analyze the genetic relationships between the strains based on the genotype of 70 Brucella isolated from different hosts in Qinghai Province. Results:A total of 1 466 blood samples of Himalayan marmot were collected from Qinghai-Tibet Plateau. Two strains of Brucella were isolated and cultured from 64 RBT-positive samples, named QH2013054 and QH2013062, respectively. They were identified as Brucella ovis biotype Ⅲ by conventional and molecular biological methods. The MLVA genotyping results showed that QH2013054 and QH2013062 were different at the Bru16 locus, indicating different MLVA genotypes. Cluster analysis showed that strain QH2013054 had the same MLVA genotype as 7 strains, among which 6 strains were from 3 farmers and 3 sheep from the same family in Gonghe County, and 1 strain was from a farmer in Menyuan Hui Autonomous County. The strain QH2013062 had the same MLVA genotype as 4 strains, including 3 strains from 3 farmers in Menyuan Hui Autonomous County and 1 strain from a farmer in Tu Autonomous County of Huzhu. Conclusions:The strains of Brucella isolated from Himalayan marmot in Qinghai-Tibet Plateau of Qinghai Province have the same MLVA genotype as some strains of Brucella isolated from humans and sheep in Qinghai Province. It is speculated that the host humans, sheep and Himalayan marmot in Qinghai-Tibet Plateau may have a common source of infection.

Objective To explore the impact of glucocorticoid on coagulation through administrating on rats with smoke inhalation.Methods Totally 150 male S-D rats were randomly (random number) divided into 5 groups:control group (ambient air inhalation),smoke group (smoke inhalation for 30 min),smoke+high dosage methyl prednisolone group(MP 40 mg/kg,intraperitoneal injection,s+HMP group),smoke+medium dosage MP (4 mg/kg) group (s+MMP group),smoke+low dosage MP (0.4 mg/kg) group (s+LMP group) (all n=30).Survival rates were calculated 24 h after smoke inhalation.Lung tissues were collected for histopathology and wet to dry (W/D) ratio.Arterial blood was collected for blood gas test.Coagulation factors in lung and plasma were tested.Results Survival rates of three MP groups were markedly improved compared with the smoke group (all P＜0.05),and was significantly higher in the medium dosage group(85.17％) than those in the low and high dosage groups (65.73％ and 60.07％,all P＜0.05).The W/D ratio and blood gas test were markedly improved in the high and medium groups (all P＜0.05).Tissue factor (TF) and thrombin-antithrombin complex (TAT-c) in bronchoalveolar lavage fluid (BALF) increased dramatically after SI (P＜0.01,P=0.005) with a remarkable drop of factor Ⅱ (F Ⅱ) (P=0.007),all of which were attenuated by MP with dosage dependence.The mRNA expression of TF increased dramatically after SI and recovered significantly with MP administration,while the expression of thrombomodulin (TM) recovered in the opposite direction with MP,all of which were in a dosage dependent manner.TF,fibrinogen (FIB),TAT-c increased significantly in plasma after smoke inhalation (P＜0.01,P=0.027,P=0.005).F Ⅷ ％ increased with MP administration and TF was raised by high dosage MP compared with the smoke group.FIB and TAT-c were decreased in all MP groups,which were significant higher in the high and middle dosage groups.The change of TM and endothelial cell protein C receptor (EPCR) in circulation were similar with FIB or TAT-c with or without MP.Protein C (PC％) and antithrombin (AT Ⅲ ％) dropped dramatically after SI,high and middle dosages of MP could restore the activity significantly,while low dosage would restore AT Ⅲ ％ but not PC％.Conclusions Glucocorticoid can significantly improve local and systemical coagulation disorder caused by smoke inhalation,and high-and medium-dosage hormones are effective.The regulation of hormones on the coagulation system is an important mechanism in the treatment of smoke inhalation induced lung injury.

This study investigated the growth-regulating effects of progesterone(Prog)on nPR-negative malignant melanoma cells and the possible mechanisms.A375 and A875 cells were cultured and treated with Prog of different concentrations.For signal transduction pathway studies,the cells were pretreated with Prog receptor antagonist(RU486,1×10 7 mol/L)or MAPK inhibitor (U0126,5×10-6 mol/L)for 1 h and then co-incubated with prog(10-9 mol/L)for another 24 h.Indirect immunofluorescence assay,MTT,flow cytornetry and Western blotting were used for assessing the nPR expression,cell growth,cell apoptosis and ERK1/2 Phosphorylation,respectively.Our results showed that lower progesterone concentration promoted the proliferation of both A375 and A875 cells,but this growth-stimulatory effect decreased at progesterone concentration of 1 × 10-7mol/L or higher.The response could be abolished by MAPK inhibitor U0126,but could not be blocked by progesterone antagonist RU486.Flow cytometry exhibited that high concentration(≥1×10-7 mol/L)progesterone increased the apoptosis of the two cells in a dose-dependent manner.The level of ERK 1/2 phosphorylation was increased by a lower progesterone concentration,but reduced by a higber concentration(1×10-6 mol/L).These results suggest progesterone exerts growth-regulating effects on nPR-negative tumor cells through a non-genomic mechanism.