OBJECTIVE: To investigate the inflammatory effects of Cinobufotalin on monocytes in resting state and macrophages in activated state and its molecular mechanism. METHODS: THP-1 cells were stimulated with Phorbol 12-myristate 13-acetate to induce differentiation into macrophages. Lipopolysaccharides was added to activate macrophages in order to establish macrophage activation model. Cinobufotalin was added to the inflammatory cell model for 24 h as a treatment. CCK-8 was used to detect cell proliferation, Annexin V /PI double staining flow cytometry was used to detect cell apoptosis, flow cytometry was used to detect macrophage activation, and cytometric bead array was used to detect cytokines. Transcriptome sequencing was used to explore the gene expression profile regulated by Cinobufotalin. Changes in the significantly regulated molecules were verified by real-time quantitative polymerase chain reaction and Western blot. RESULTS: 1∶25 concentration of Cinobufotalin significantly inhibited the proliferation of resting monocytes(P<0.01), and induced apoptosis(P<0.01), especially the activated macrophages(P<0.001, P<0.001). Cinobufotalin significantly inhibited the activation of macrophages, and significantly down-regulated the inflammatory cytokines(IL-6, TNF-α, IL-1β, IL-8) released by activated macrophages(P＜0.001). Its mechanism was achieved by inhibiting TLR4/MYD88/P-IκBa signaling pathway. CONCLUSION Cinobufotalin can inhibit the inflammatory factors produced by the over-activation of macrophages through TLR4/MYD88/P-IκBa pathway, which is expected to be applied to the treatment and research of diseases related to the over-release of inflammatory factors.
Humans ; Toll-Like Receptor 4/metabolism* ; Myeloid Differentiation Factor 88/genetics* ; Macrophages/metabolism* ; Cytokines/metabolism* ; Lipopolysaccharides/pharmacology* ; NF-kappa B

OBJECTIVES: To investigate the value of CCKBR^fl/fl villin-Cre mice as a mouse model of salt-sensitive hypertension (SSH). METHODS: In the first part, 2-month-old CCKBR^fl/fl villin-Cre mice (CKO) and control CCKBR^fl/fl mice (WT) were fed with normal diet (0.4% NaCl) or high salt diet (4% NaCl), separately for 6 weeks. In the rescue study, one week of hydrochlorothiazide or saline injection were treated with the CKO mice fed high salt diet. The blood pressure, biochemical indexes, and the expression of small intestinal sodium transporters (NHE3, NKCC1, eNaC) was detected. The organ injury markers (MMP2/MMP9) and the histopathological changes of kidneys were observed, whereas the changes of duodenal sodium absorption were detected by small intestinal perfusion in vivo. RESULTS: The CCKBR^fl/fl villin-Cre mice with high salt intake exhibited high blood pressure, increased duodenal sodium absorption and urinary sodium excretion, and with renal injury. The protein expression of NHE3, NKCC1 and eNaC were also significant increase in the intestine of CKO-HS mice. Treatment with hydrochlorothiazide remarkably attenuated the elevated blood pressure by high salt absorption in the CCKBR^fl/fl villin-Cre mice, but no significant histopathological changes were observed. CONCLUSIONS These results support a crucial role of intestinal Cckbr deficiency on SSH development and the diuretic antihypertension effect in CCKBR^fl/fl villin-Cre mice. The CCKBR^fl/fl villin-Cre mice with the high salt intake may serve as a stable model of salt-sensitive hypertensive induced by sodium overloading.

Platelets are reprogrammed by cancer via a process called education, which favors cancer development. The transcriptional profile of tumor-educated platelets (TEPs) is skewed and therefore practicable for cancer detection. This intercontinental, hospital-based, diagnostic study included 761 treatment-naïve inpatients with histologically confirmed adnexal masses and 167 healthy controls from nine medical centers (China, n = 3; Netherlands, n = 5; Poland, n = 1) between September 2016 and May 2019. The main outcomes were the performance of TEPs and their combination with CA125 in two Chinese (VC1 and VC2) and the European (VC3) validation cohorts collectively and independently. Exploratory outcome was the value of TEPs in public pan-cancer platelet transcriptome datasets. The AUCs for TEPs in the combined validation cohort, VC1, VC2, and VC3 were 0.918 (95% CI 0.889-0.948), 0.923 (0.855-0.990), 0.918 (0.872-0.963), and 0.887 (0.813-0.960), respectively. Combination of TEPs and CA125 demonstrated an AUC of 0.922 (0.889-0.955) in the combined validation cohort; 0.955 (0.912-0.997) in VC1; 0.939 (0.901-0.977) in VC2; 0.917 (0.824-1.000) in VC3. For subgroup analysis, TEPs exhibited an AUC of 0.858, 0.859, and 0.920 to detect early-stage, borderline, non-epithelial diseases and 0.899 to discriminate ovarian cancer from endometriosis. TEPs had robustness, compatibility, and universality for preoperative diagnosis of ovarian cancer since it withstood validations in populations of different ethnicities, heterogeneous histological subtypes, and early-stage ovarian cancer. However, these observations warrant prospective validations in a larger population before clinical utilities.
Humans ; Female ; Blood Platelets/pathology* ; Biomarkers, Tumor/genetics* ; Ovarian Neoplasms/pathology* ; China

OBJECTIVES: To investigate the clinical treatment outcomes and the changes of the outcomes over time in extremely preterm twins in Guangdong Province, China. METHODS: A retrospective analysis was performed for 269 pairs of extremely preterm twins with a gestational age of <28 weeks who were admitted to the department of neonatology in 26 grade A tertiary hospitals in Guangdong Province from January 2008 to December 2017. According to the admission time, they were divided into two groups: 2008-2012 and 2013-2017. Besides, each pair of twins was divided into the heavier infant and the lighter infant subgroups according to birth weight. The perinatal data of mothers and hospitalization data of neonates were collected. The survival rate of twins and the incidence rate of complications were compared between the 2008-2012 and 2013-2017 groups. RESULTS: Compared with the 2008-2012 group, the 2013-2017 group (both the heavier infant and lighter infant subgroups) had lower incidence rates of severe asphyxia and smaller head circumference at birth (P<0.05). The mortality rates of both of the twins, the heavier infant of the twins, and the lighter infant of the twins were lower in the 2013-2017 group compared with the 2008-2012 group (P<0.05). Compared with the 2008-2012 group, the 2013-2017 group (both the heavier infant and lighter infant subgroups) had lower incidence rates of pulmonary hemorrhage, patent ductus arteriosus (PDA), periventricular-intraventricular hemorrhage (P-IVH), and neonatal respiratory distress syndrome (NRDS) and a higher incidence rate of bronchopulmonary dysplasia (P<0.05). CONCLUSIONS There is a significant increase in the survival rate over time in extremely preterm twins with a gestational age of <28 weeks in the 26 grade A tertiary hospitals in Guangdong Province. The incidences of severe asphyxia, pulmonary hemorrhage, PDA, P-IVH, and NRDS decrease in both the heavier and lighter infants of the twins, but the incidence of bronchopulmonary dysplasia increases. With the improvement of diagnosis and treatment, the multidisciplinary collaboration between different fields of fetal medicine including prenatal diagnosis, obstetrics, and neonatology is needed in the future to jointly develop management strategies for twin pregnancy.
Bronchopulmonary Dysplasia/epidemiology* ; Female ; Gestational Age ; Humans ; Infant ; Infant, Extremely Premature ; Infant, Newborn ; Pregnancy ; Respiratory Distress Syndrome, Newborn/epidemiology* ; Retrospective Studies ; Treatment Outcome

Objective To investigate the effects of Smad7 knock down by lentivirus on rat cardiac fibroblasts proliferation, migration, cell differentiation and collagen secretion in vitro. Methods The primary cardiac fibroblasts were separated from the hearts of ten SD rats and identified by immunohistochemical method. The lentivirus transfection knocked down the expresson of Smad7 in cardiac fibroblasts, Western blotting was used to detect the efficiency of Smad7 knock down by lentivirus. The proliferation of cardiac fibroblasts was quantified by real-time unlabeled cell analyzer. Cell migration was evaluted by cell wound scratch assay. Western blotting was used to detect expression of α-smooth muscle actin（α-SMA） and collagen Ⅰ（Col Ⅰ）. Results Myocardial fibroblasts were successfully cultured and identified by immunocytochemical methods. The multiplicity of infection（MOI） that lentivirus transduction of myocardial fibroblasts was 100. After lentivirus transduction, 88.33% myocardial fibroblasts expressed green fluorescent protein, showed that the lentivirus could significantly reduce the protein expression of Smad7. Smad7 deficiency decreased the proliferation and migration of cardiac fibroblasts, increased the protein expression of α-SMA and decreased collagen secretion. The results indicated that Smad7 deficiency significantly down-regulated the proliferation and migration of cardiac fibroblasts, increased α-SMA protein expression and reduced ColⅠ protein expression. Conclusion Smad7 deficiency can significantly change the cardiac fibroblasts function, that is related to the pathological mechanism that lead to myocardial fibrosis

Dry eye is a multifactorial disease, which seriously affect the work and life of patients. Since the pathogenesis of dry eye has not been clarified so far, there is no standard animal model available now. Non-Sjögren syndrome dry eye is one of the main types of dry eye, which requires further research. Studies on dry eye mechanisms have led to the development and improvement of animal models. In this paper, the methods of animal models of non-Sjögren syndrome dry eye, including drug treatments, surgery, exogenous injury, behavioral patterns, changes in diet, are reviewed. In addition,different models are described, the problems are analyzed and perspectives and reflections on future research are offered.

OBJECTIVE: To verify the clinical effect of acupuncture on knee osteoarthritis (KOA). METHODS: Forty-two patients with KOA were randomly divided into an acupuncture group (21 cases, 1 case dropped off) and a sham acupuncture group (21 cases, 1 case dropped off). The patients in the acupuncture group were treated with routine acupuncture at 5-6 local acupoints [Dubi (ST 35), Neixiyan (EX-LE 4), Heding (EX-LE 2), Yinlingquan (SP 9), Xuehai (SP 10), Zusanli (ST 36), etc.] and 3-4 distal acupoints [Fengshi (GB 31), Waiqiu (GB 36), Xuanzhong (GB 39), Zulinqi (GB 41), etc.]. The patients in the sham acupuncture group were treated with shallow needling technique at non-acupoint. The needles were retained for 30 min in both groups. All the treatment was given three times a week for 8 weeks. Knee injury and osteoarthritis outcome score (KOOS) were recorded before and after treatment and 18-week follow-up. RESULTS: Compared before treatment, the scores of 5 dimensions of KOOS [pain, symptoms (except pain), daily activities, sports and entertainment, and quality of life] were increased after treatment and during follow-up in the two groups ( CONCLUSION Acupuncture can reduce the pain symptoms and improve daily activities in patients with KOA.
Acupuncture Points ; Acupuncture Therapy ; Humans ; Knee Injuries ; Osteoarthritis, Knee/therapy* ; Quality of Life ; Treatment Outcome

Pilot study is essential prerequisite for large-scale research. Acupuncture is a complex intervention measure. The factors of its clinical effect and effect mechanism are complicated. Hence, the resource waste is easily induced in research if the large-scale clinical research is rashly carried out before scientific verification. Currently, there is still a lack of high-quality evidence of clinical research, which affects the promotion and application of the curative effect of acupuncture. The pilot study of acupuncture can evaluate the feasibility and the process coordination of trial, optimize program and process and provide the basis of sample size calculation prior to launching a full-scale trial. Thereby, the methodological quality of acupuncture research and the reliability of trial results can be improved. In the paper, the arguments focus on the definition, design and report of pilot study so as to provide the approaches and references for the design and implementation of pilot study of clinical research of acupuncture.
Acupuncture Therapy ; Pilot Projects ; Reproducibility of Results ; Research Design ; Treatment Outcome

OBJECTIVE: To evaluate the clinical effect of acupuncture for the improvements in various dyspeptic symptoms of postprandial distress syndrome. METHODS: The secondary analysis on the data of a multi-center randomized controlled trial (RCT) was conducted. 278 patients with postprandial distress syndrome were randomized into an acupuncture group (138 cases) and a sham-acupuncture group (140 cases). In the acupuncture group, acupuncture was applied to Baihui (GV 20), Danzhong (CV 17), Zhongwan (CV 12), Qihai (CV 6), Neiguan (PC 6), Zusanli (ST 36), etc. In the sham-acupuncture group, 6 sites, neither located on meridians nor belonged to meridian acupoints, were selected and punctured shallowly. The duration of treatment was 20 min each time, 3 times a week, for 4 weeks totally in the two groups. The follow-up visit lasted for 12 weeks. The scores of dyspeptic symptoms were compared between the two groups before treatment, during treatment (in week 1, 2, 3 and 4) and during follow-up (in week 8, 12 and 16) separately. RESULTS: Besides the scores of early satiety and vomiting in the sham-acupuncture group in week 1, the scores of the other dyspepsia symptoms during treatment and follow-up were all reduced in the two groups as compared with those before treatment ( CONCLUSION Acupuncture remarkably relieves postprandial fullness, early satiety, upper abdominal bloating and belching in patients with postprandial distress syndrome.
Acupuncture Points ; Acupuncture Therapy ; Dyspepsia/therapy* ; Humans ; Meridians ; Treatment Outcome

Objective To examine the effect of the integrated schistosomiasis control measures in Hunan Province from 2004 to 2019, so as to provide insights into the development of the schistosomiasis elimination strategy. Methods The integrated schistosomiasis control measures implemented by the health, agriculture, water resources, forestry, land and resources sectors were retrospectively collected in Hunan Province from 2004 to 2019, and the completion of each measure, cost of control measures, Schistosoma japonicum infections in humans and bovines, and snail status were analyzed each year. An index system for assessing the integrated schistosomiasis control effect was constructed using the Delphi method to calculate the integrated schistosomiasis control effect index. In addition, a cost-effect analysis was performed in terms of the decline in the prevalence of S. japonicum infections in humans and bovines, areas with snails in inner embankments, and areas with infected snails. Results A total of 7 110 926 500 Yuan were invested into the integrated schistosomiasis control program of Hunan Province from 2004 to 2019. During the 16-year period, a total of 277 437.12 hm² snail habitats received molluscicidal treatments, 6 927 230 person-times given expanded chemotherapy, 2 116 247 bovine-times given expanded chemotherapy, 954 850 harmless toilets built, 290 359 bovines fenced, 136 666 bovines eliminated, 141 905 machines used to replace bovines, 39 048.63 hm² water lands improved as dry lands, 724.12 km irrigation regions improved, 3 994 300 populations covered with safe water, 191 102.89 hm² forests planted and 38 535.27 hm² lands leveled. The prevalence of S. japonicum infections was 4.29% in humans and 4.48% in bovines in Hunan Province in 2004, with 2 449.37 hm² snail habitats in inner embankments and 3 423.74 hm² infected snail areas. In 2019, the prevalence of S. japonicum infections reduced to 0 in both humans and bovines, and areas of snail habitats reduced to 540.92 hm² (77.92% reductions), while the areas with infected snails reduced to 0. The overall integrated schistosomiasis control effect index appeared a tendency towards a rise over years since 2004, and the integrated schistosomiasis control effect index was 97.35 in 2019; the annual mean costs for a 1% reduction in the prevalence of S. japonicum infections in 100 populations and 100 bovines were 70.11 Yuan and 4 204.78 Yuan, and the annual mean costs for a 1% reduction in the snail areas in inner embankments and infected snail areas were 2 010.20 Yuan and 1 298.09 Yuan, respectively. Conclusions The integrated control measures achieve remarkable effectiveness for schistosomiasis control in Hunan Province, with a remarkable decline in the prevalence of S. japonicum infections in humans and bovines and great shrinking of snail areas in inner embankments and infected snail areas. Adequate fund investment is required to improve the integrated schistosomiasis control measures and consolidate the control achievements.