Objective To investigate the changes in myeloid-derived suppressor cells (MDSC), regulatory T cells (Treg) and T helper 17 (Th17) cells in peripheral blood of patients with HPV infection and cervical diseases including ectopic cervical columnar epithelium (CE), cervical intraepithelial neoplasia (CIN) and cervical squamous cell carcinoma (CC), and to analyze the roles of MDSC, Treg and Th17 cells in the development of cervical diseases.Methods This study enrolled 120 HPV-positive patients with cervical diseases (18 cases of CE, 50 cases of CIN, 52 cases of CC), 20 HPV-negative patients with cervical diseases (10 cases of CE and 10 cases of CIN) and 20 healthy subjects from March 2011 to December 2016.Changes in MDSC, Treg and Th17 cells in peripheral blood of all patients were detected by flow cytometry.Results Percentages of MDSC, Treg and Th17 cells in peripheral blood of patients who were positive for HPV were significantly higher than those in HPV-negative patients (P<0.05).Besides, percentages of these cells gradually increased with the exacerbation of cervical disease (CE-CINⅠ-CINⅡ-CINⅢ-CC) (P<0.05).Patients who were infected with high-risk HPV had significantly higher percentages of MDSC, Treg and Th17 cells in peripheral blood than those infected with low-risk HPV (P<0.05).Changes of MDSC, Treg and Th17 cells in peripheral blood of HPV-positive patients with cervical cancer were related to International Federation of Gynecology and Obstetrics (FIGO) stage, tumor differentiation, lymph node metastasis and vascular invasion (P<0.05).MDSC, Treg and Th17 cells in peripheral blood of all patients were significantly reduced after treatment (P<0.05).Patients whose status changed from HPV-positive to HPV-negative following treatment also showed significantly decreased MDSC, Treg and Th17 cells in peripheral blood as compared with those who remained HPV-positive (P<0.05).MDSC, Treg and Th17 cells in peripheral blood of HPV-positive patients with cervical diseases were positively correlated (r=0.599, 0.677, 0.648;P<0.05).Conclusion MDSC, Treg and Th17 cells in peripheral blood of patients with HPV infection are associated with the pathological process, severity and clinical pathological features of cervical diseases as well as therapeutic effects.Therefore, they can be used as biomarkers to detect the occurrence and development of HPV-positive cervical diseases.

Objective To evaluate the effect of calcitonic gene related peptide(CGRP) and endothelin(ET) on acute and delayed cerebral vasospasm(CVS) after subarachnoid hemorrhage(SAH).Methods The SAH model in rabbit were established by single injection of autologous arterial blood(0.5 ml/kg) to cisterna magna on percutaneous suboccpital route. Transcranial Dopper(TCD) was performed to detect the spastic of rabbit basilar artery(BA). The CGRP and ET concentrations in cerebrospinal fluid were examinated quntitatively by radioimmunologlc techniques at 30 minutes, 3 days, and 5 days after SAH.Results The results of TCD showed that rabbit basilar artery was spastic at 30 minutes, 3 days and 5 days after SAH. At 30 minutes after SAH, CGRP concentration significantly increased upto about 18 fold( P

Objective To evaluate the value of N-terminal prohormone brain natriuretic peptide (NT-proBNP) and tissue Doppler imaging (TDI) in patients with left ventricular diastolic dysfunction.Methods Forty-three patients with diastolic heart failure were enrolled. Pulsed wave (PW) Doppler technology was used to detect early diastolic mitral velocity (E) and TDI was performed to determine the mitral annulus velocity of early diastolic phase (Em). According to the E/Em, patients were divided into three groups: group Ⅰ (E/Em ＜8),group Ⅱ (E/Em 8-15) and group Ⅲ (E/Em ＞ 15). The plasma NT-proBNP was measured. Results The plasma NT-proBNP in group Ⅲ [( 1119 ± 3123) ng/L] was higher than that in group Ⅱ [(684± 1742) ng/L] and group Ⅰ [(276±438) ng/L] (P＜0.01),and the plasma NT-proBNP in group Ⅱ was higher than that in group Ⅰ (P ＜ 0.01 ). The plasma NT-proBNP had positive correlation with E/Em (r = 0.438,P ＜ 0.01 ). Conclusion The plasma NT-proBNP and TDI are useful to evaluate the left ventricular diastolic dysfunction.

The aim of this study is to identify the express specificity of bone morphogenetic protein 15 (Bmp15) in porcine. The pBMP15-EGFP reporter vector was constructed from the 2.2 kb fragment of porcine bmp15 promoter to trace the differentiation process of stem cells into oocyte-like cells. We used porcine ovary and Chinese Hamster Ovary cell line (CHO), mouse myoblast cell line (C2C12) and porcine amniotic fluid stem cell (pAFSC) to investigate the expression and regulation of this gene via RT-PCR, immunofluorescence, cell transfection, and microinjection methods. We also used single layer cell differentiation to detect the application potential of bmp15. The results show that bmp15 gene was specifically expressed in the porcine ovary and CHO rather than in C2C12 and pAFSC. In addition, the characteristic of tissue-specific of Bmp15 was detected on CHO instead of other cell lines by transient transfection. We also detected the expression of Bmp15 in oocyte at different development stages by immunofluorescence of fixed paraffin-embedded ovary sections. Furthermore, microinjection results show that bmp15 expressed in oocytes at 18 h of maturation in vitro, and continued up to 4-cell stage embryos. Most importantly, we found that the expression of Bmp15 started at day 12 after inducing pAFSC into oocyte-like cells by transfection; green fluorescent was visible in round cell masses. It indicated that bmp15 has the expression specificity and the pBMP15-EGFP reporter vector can be used to trace Bmp15 action in the differentiation of stem cells into germ cells.
Animals ; Bone Morphogenetic Protein 15 ; genetics ; CHO Cells ; Cell Differentiation ; Cricetinae ; Cricetulus ; Female ; Genes, Reporter ; Genetic Vectors ; Mice ; Microinjections ; Myoblasts ; cytology ; Oocytes ; metabolism ; Ovary ; metabolism ; Stem Cells ; cytology ; Swine

This study aims to investigate the preventive role and potential mechanisms of blocking extracellular HMGB1 function on doxorubicin induced cardiac injury. Mice were treated with HMGB1 blocker glycyrrhizin 1 h before and one time every day (intraperitoneal, 10 mg per mouse) after doxorubicin injection, and sacrificed on the day 14 after doxorubicin challenge. Cardiac function was evaluated by echocardiography and hemodynamic measurement. Myocardial inflammation and collagen deposition were analyzed by immunohistochemistry and picrosirius red staining. The interaction of HMGB1 and TLR2 was assessed by co-immunoprecipitation and confocal microscopy. The protein contents of HMGB1, MyD88, p65NF-kappaB and phospho-p65NF-kappaB were measured by Immunoblot. Compared with mice treated with saline, doxorubicin treatment led to an upregulation in HMGB1 expression. Blocking HMGB1 activity with glycyrrhizin protected mice against cardiac dysfunction, inflammatory response, and cardiac fibrosis induced by doxorubicin challenge. Glycyrrhizin inhibited the interaction of HMGB1 and TLR2, and blocked the downstream signaling of TLR2. In conclusion, blocking HMGB1 protected against doxorubicin induced cardiac injury by inhibiting TLR2 signaling pathway.

Oocyte-like cells (OLC) can be generated by stem cells after the induction and differentiation in vitro, and maturated when transplanted in vivo to improve the development potential. Human amniotic fluid stem cells (hAFSC) were cultured for 10 days in porcine follicle fluid (pFF) that was extracted from the medium follicle with high levels of hormones and Bmp 15 protein. After the induction, the cell aggregates showed the germ cell-like cells and produced the germ cell marker oct4, and triggered epigenetic changes with high expression of methylation transferase gene dnmt3b. The cell aggregates were packaged into porcine theca folliculi to form grafts, which were then transplanted into mouse renal capsule. After one month of transplantation, the morphology of OLC from a graft was not only similar to oocytes, but also expressed the germ cells markers (oct4, nanog, stella, ifitm3, dazl, nanos3, bmp15, and gd9). The results demonstrate that the in vivo differentiation model was useful for OLC development.
Amniotic Fluid ; cytology ; Animals ; Biomarkers ; Cell Differentiation ; Female ; Humans ; Mice ; Oocytes ; cytology ; Ovarian Follicle ; Stem Cells ; cytology ; Swine

Since the first local infection case of Zika virus appeared in Brazil from May 2015, the disease quickly spread in Brazil. The writer would introduce Zika virus from 4 aspects and they are history, biological property, physicochemical property and physicochemical property. Combined the writer′s years of clinical nursing experience in tertiary hospitals, experience during Ebola period and study on Ebola, this paper would analyze how to nursing patients with this disease from 5 aspects; it also put forward some suggestions on prevention and control of Zika virus disease from aspect which includes society, government, hospitals and the public, combined with the prevention and control of infectious disease, so that to make everyone realize the importance of learning, prevention and control of epidemic, nursing, standard nursing operation and popularized knowledge of infectious diseases.

Objective:To explore the characteristics and changes of cardiac injury with age in Duchenne muscular dystrophy (DMD) and its clinical significance.Methods:A prospective cohort study was conducted. The 215 patients diagnosed with DMD in West China Second Hospital from January 2019 to November 2022 and aged from 6 to 18 years were enrolled. Their clinical data, myocardial injury markers, routine electrocardiogram, cardiac magnetic resonance (CMR) and echocardiography were collected. The patients were divided into five age groups: 6-<8, 8-<10, 10-<12, 12-<14 and 14-18 years of age, and matched with healthy boys respectively. Independent sample t test or Mann-Whitney U test was used to compare the clinical data and CMR indexes between DMD patients and controls in all age subgroups, and to compare the value of left ventricular ejection fraction (LVEF) measured by echocardiography and CMR in each subgroup of DMD patitents. Pearson correlation analysis or Spearman correlation analysis was used to explore the relation between the CMR indexes and age in DMD patients. Results:A total of 215 patients with DMD (all male) and 122 healthy boys were included in the study. There were 75 DMD patients and 23 controls in 6-<8 years of age group, 77 DMD and 28 controls in 8-<10 years of age group, 39 DMD and 23 controls in 10-<12 years of age group, 10 DMD and 31 controls in the 12-<14 years of age group, and 14 DMD and 17 controls in 14-18 years of age group. In the DMD patients, the older the age, the lower the levels of creatine kinase (CK) and creatine kinase isoenzyme (CK-MB). In the 6-<8 years of age group, the CK level was 10 760 (7 800, 15 757) U/L, while in the group of 14-18 years of age, it was 2 369 (1 480, 6 944) U/L. As for CK-MB, it was (189±17) μg/L in the 6-<8 years of age group and (62±16) μg/L in the 14-18 years of age group. Cardiac troponin I remained unchanged in <12 years of age groups, but significantly increased in 12-<14 years of age group, reaching the highest value of 0.112 (0.006, 0.085) μg/L. In the DMD patients, the older the age, the higher the proportion of abnormal electrocardiogram (ECG). In the 6-<8 years of age group, the proportion is 29.3% (22/75), while in the 14-18 years of age group, it was 10/14. Correlation analysis showed that the left ventricular end-diastolic volume index was positively related with age ( r=0.18, P=0.015), and the left ventricular stroke volume index and cardiac output index were negatively related with age ( r=-0.34 and -0.31, respectively, both P<0.001). In the DMD patients, the older the age, the lower LVEF, with the LVEF decreasing to (49.3±3.1)% in the 14-18 years of age group. The LVEF of DMD cases was significantly lower than that of controls in the age subgroups of 8-<10, 10-<12, 12-<14 and 14-18 years of age groups ((57.9±5.2) % vs. (63.6±0.8)%, 60.7% (55.9%, 61.9%) vs. 63.7% (60.2%, 66.0%), 57.1% (51.8%, 63.4%) vs. 62.1 % (59.5%, 64.5)%, (49.3±3.1) % vs. (61.6±1.3)%, respectively; all P<0.01). In the DMD patients, the older the age, the higher the proportion of positive late gadolinium enhancement (LGE). In the 6-<8 years of age group, it was 22% (11/51), in the 12-<14 years of age group, it was 13/14, and in the 14-18 years of age group, all DMD showed positive LGE. The value of LVEF of DMD cases measured by echocardiography was significantly higher than that measured by CMR in 6-<8 years of age group and 8-<10 years of age group (63.2% (60.1%, 66.4%) vs. 59.1 % (55.4%, 62.9%), and (62.8±5.2) % vs. (57.9±5.2)%, all P<0.001). Conclusion:DMD patients develop cardiac injury in the early stage of the disease, and the incidence of cardiac damage gradually increases with both age and the progression of disease.

Laminin (LN) proteins are important components of extracellular matrix. These proteins regulate cell proliferation, differentiation, migration, and tissue repair. The LN family has 12 genes that encode 5 α, 4 β, and 3 γ proteins. LamininA5 (LAMA5) as an important gene can support pluripotent cell growth and have been widely studied. However, porcine LAMA5 is absent in all tested porcine genomic databases so far. In this study, we confirmed for the first time the existence of porcine LAMA5 through bioinformatics analysis, and verified this result by cDNA cloning and sequencing. To reveal the expression pattern of Laminin gene family, we detected the expression of Laminin genes in porcine tissues, somatic cells, and porcine induced pluripotent stem cells (piPSCs). The results showed that an alternative splicing variant of Laminin B1 (LAMB1-a) was found exclusively in all tested piPSCs. The expression of this alternative splicing variant is positively correlated with the pluripotent state of piPSCs. The above findings provide evidences and foundations for the father use of LN as extracellular matrix to facilitate the derivation and culture of porcine pluripotent stem cells.

Objective To analyze the application value of uterine artery flow and cerebral placental rate(CPR)in diagnosing fetal growth restriction(FGR).Methods A total of 114 pregnant women with clinically diagnosed late-onset FGR who were hospitalized in Jiangxi Maternal and Child Health Hospital from January 2021 to June 2022 were assigned to study group,and 122 pregnant women with normal intrauterine development were assigned to control group.The blood flow parameters of uterine artery(UtA),umbilical artery(UA)and middle cerebral artery(MCA)in two groups were determined by ultrasound,and CPR in two groups was calculated.The blood flow difference and pregnancy outcome of two groups were compared.Receiver operating characteristic(ROC)curve was used to analyze the application value of UtA and CPR alone and combined in the clinical diagnosis of FGR.Results The UtA resistance index(RI)of pregnant women in study group was higher than that of control group,the fetal UA blood flow parameter was higher than that of control group,the MCA blood flow parameter and the CPR value were both lower than those of control group,the differences were statistically significant(P<0.05).The birth weight and 1min Apgar score of study group were lower than those of control group(P<0.001).In addition,the incidence of emergency cesarean section operation,premature delivery and neonates transferred to neonatal intensive care unit(NICU)due to various complications in study group were significantly higher than those in control group(P<0.05).ROC curve showed that in predicting FGR,the area under the curve(AUC)of UtA-RI was 0.82(95%CI:0.77-0.88).The predictive efficiency of CPR was 0.75(95%CI:0.69-0.81).The combination of UtA-RI and CPR parameters had the highest efficiency in predicting FGR,with an AUC of 0.92(95%CI:0.89-0.95).Conclusion CPR combined with UtA-RI monitoring has clinical application value for early detection of FGR,guiding intervention,and improving adverse perinatal outcomes.