A screening of thyroid function was performed in 2 320 pregnant women whose gestational age was less than 12 weeks.The incidence of subclinical thyroid disease and its relationship with gestational diabetes mellitus were analyzed.The results showed that there were 8 patients (0.34％) with clinical hypothyroidism,226 (9.74％) with subclinical hypothyroidism,and 14 patients (0.6％) with hyperthyroidism among 2 320 pregnant women.The prevalence of gestational diabetes mellitus in pregnant women with subclinical hypothyroidism was apparently higher than that in normal pregnant women (33.3％ vs 12.9％,P＜0.05),suggesting that the increased incidence of gestational diabetes mellitus seems to be related to subclinical hypothyroidism during early pregnancy.Therefore,it is important to screen the function of thyroid during early pregnancy

Aim To observe the expression of MRTF-A in rat glomerular mesangial cells(GMCs) induced by advanced glycation end products(AGEs) and its effect on ICAM-1 and FN;to explore whether MRTF-A is involved in the process of diabetic nephropathy by affecting NF-κB pathway.Methods Under the condition of AGEs, CCG-1423 and anti-MRTF-A small interfering RNA were used to knock down MRTF-A and MRTF-A plasmid was used to activatt MRTF-A, The expression level of MRTF-A, ICAM-1, FN and p65 in nucleus were detected by Western blot.Results The protein expressions of MRTF-A was increased in AGEs-induced GMCs.The expressions of FN and ICAM-1 and p65 in nucleus were downregulated by knocking down MRTF-A.However, the expressions of FN, ICAM-1 and p65 in nucleus were upregulated by overexpressing MRTF-A.Conclusions AGEs can upregulate the expression of MRTF-A in GMCs, and MRTF-A mediates the protein expressions of FN and ICAM-1 by affecting NF-κB signaling pathway in AGEs-induced GMCs.

OBJECTIVE To explore the expression of Bcl-2, on mRNA and protein levels, in the different age of C57BL/6 mice cochleae and the expression localization in the cochleae.METHODS Using ABR to test the hearing level in C57BL/6 mice. Surface preparation of cochlear basil membrane is used to observe the morphology and amout of the outer and inner hair cells in different age of C57BL/6 mice. Fluorescent quantitative real time PCR, immunofluorescence histochemical method and western blot are used to detect the expression of Bcl-2 on the mRNA and protein levels in the C57BL/6 mice cochlea of different age groups ('young group', 'elderly group').RESULTS ABR results showed that the hearing threshold of 'older' C57BL/6 mice is much higher than that in the 'young' mice, and surface preparation of cochlear basil membrane showed the hair cell localized in the cochlear basil turn of 'old' mice arranged in a disorder station and part of hair cells were lost. Also, the spiral ganglion cells arranged sparsely and messily. Fluorescence quantitative real-time PCR results suggest the expression of Bcl-2 on/at the mRNA level of the 'old' mice cochleae decreases significantly, compared to that in the 'young' mice. The results of Immunofluorescence and Western blot suggest the expression of Bcl-2 on/at the protein levels of the 'old' mice cochlea decreased, compared to that in the 'young' mice. Also, the Bcl-2 is located in the cytoplasm, and the expression of Bcl-2 in the inner hair cells seems higher than that in the outer hair cells. CONCLUSION The expression of Bcl-2 significantly deceased in the 'old' C57BL/6 mice cochleae, both on mRNA and protein level, which may be related to the hearing loss and loss of hair cells.

ObjectiveTo observe the effect of topical sulfotanshinone sodium(STS) on sebaceous hyperplasia in animal models. MethodsThe sebaceous gland spots of adult male Syrian hamster flank organ served as the animal model. Sulfotanshinone sodium(0.5％) was applied to sebaceous gland spots in the right flank organ thrice daily, while those in the left were treated with normal saline as control. Parameters were examinedbefore, 10 days, 20 days and 30 days after the beginning of the topical treatment. A vernier caliper was utilized to measure the size of sebaceous gland spots, hematoxylin and eosin(HE) staining to observe the structure of sebaceous glands, immunohistochemistry to determine the expression of proliferating cell nuclear antigen (PCNA) in sebaceous gland cells, terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL) assay to assess the apoptosis of sebaceous gland cells. ResultsAt the baseline, no significant difference was observed in the size of sebaceous gland spots or in the proliferation and apoptosis of sebaceous gland cells between the two sides of flank organ(all P ＞ 0.05), with tightly arranged intact sebaceous glands. Compared with normal saline, sulfotanshinone sodium significantly reduced the size of sebaceous gland spots(P ＜ 0.05). Sebaceous glands were loosely arranged with decreased quantity and volume and obviously atrophic on day 30 in the right flank organ of hamsters. A decrease was observed in the expression of PCNA in sulfotanshinone sodium treated sebaceous gland cells compared with those treated with normal saline(P ＜ 0.01 ), which was more striking on day 10 and 20(both P ＜ 0.005). Sulfotanshinone sodium also induced an enhancement of apoptosis in sebaceous gland cells (P ＜ 0.01 ), which was more apparent on day 20 (P ＜ 0.005 ), and the degree of apoptosis was higher in the central area than in the peripheral area of sebaceous glands. ConclusionSulfotanshinone sodium can reduce the size and alter the microstructure of sebaceous gland spots, and inhibit the hyperplasia of sebaceous glands.

Objective To determine the effect of GnRH Ⅰ and GnRH Ⅱ on the secretion of VEGF by eutopic and ectopic endometrial stromal cells cultured in vitro, and to provide theoretical basis for exploring new treatments for endometriosis (EMs).Methods Eutopic and ectopic endometrium stromal cells cultured in vitro were treated with different concentrations of GnRH Ⅱ and a GnRH I(goserelin), and a control group was not treated by GnRH Ⅱ and GnRH Ⅰ. Enzyme linked immunosorbent assay (ELISA) was used to measure the content of vascular endothelial growth factor (VEGF) protein in the medium of the above 2 groups.Results (1)There was no difference in the VEGF protein secreted by eutopic and ectopic stromal cells in the medium after being cultured in vitro for 48 h (P＞0.05).(2)10-10, 10-8, and 10-6mol/L GnRH Ⅱ dose-dependently reduced VEGF protein secreted by endometrial stromal cells (P＜0.05),and the inhibition effect was stronger than that of GnRH Ⅰ (P＜0.05).(3)The inhibition effect of GnRH Ⅱ on VEGF in ectopic stromal cells was stronger than that of eutopic stromal cells (P＜0.05).Conclusion (1)Ectopic stromal cells cultured in vitro can secrete VEGF,which has no difference from the eutopic stromal cells, and which may play an important role in the formation and development of EMs.(2)GnRH Ⅱ can dose-dependently reduce VEGF protein secreted by ectopic and eutopic endometrial stromal cells cultured in vitro,and the inhibition effect is stronger than that of GnRH Ⅰ, providing theoretical basis for exploring new treatments for EMs.

Obective To evaluate the effects of dihydrotestosterone (DHT) on the expression of sterol regulatory element-binding protein-1c (SREBP-1c) in human HaCaT keratinocytes.Methods HaCaT cells were cultured in vitro and classified into 4 groups,i.e.,control group receiving no treatment,DIIT group treated with 3 different concentrations (10,100,1000 nmol/L) of DHT,LY294002 plus DHT group treated with DHT of 100 nmol/L after 40-minute pretreatment with the PI3K inhibitor LY294002 of 50 μmol/L,PD98059 plus DHT group treated with DHT of 100 nmol/L after 40-minute pretreatment with the MEK inhibitor PD98059 of 50 μmol/L.After another 24-hour culture,real time PCR and Western blot were carried out to detect the expression of SREBP-1c mRNA and protein in HaCaT cells,respectively.Western blot was also performed to determine the phosphorylation levels of protein kinase B (AKT),extracellular signal-regulated kinase (ERK),p38 mitogen activated protein kinase and c-Jun N-terminal kinase (JNK) in the HaCaT cells.Results DHT could enhance the expression of SREBP-1c mRNA and protein in HaCaT cells in a concentration-dependent manner,and induce the phosphorylation of AKT and ERK,but not that of P38 or JNK.The expressions of SREBP-1c mRNA and protein were significantly decreased in HaCaT cells treated with LY294002 plus DHT (7.4780 ± 1.2638 vs.21.6170 ± 2.2759,t =9.406,P ＜ 0.05; 0.7113 + 0.0313 vs.2.2577 + 0.0601,t =39.498,P ＜ 0.05),but experienced no statistical changes in those treated with PD98059 and DHT(both P ＞ 0.05),compared with those treated with DHT only.Conclusion DHT can induce the expression of SREBP-1c mRNA and protein in HaCaT cells,likely via the PI3K/AKT signaling pathway.

Objective: : To explore the effect of PD-1 gene knockout by CRISPR/Cas9 system on the proliferation and IFN-γ secretion in human T cells. Methods: : The sequence of sgRNA targeting PD-1 was designed. The PD-1-sgRNA and Cas9 mRNA were synthesized by T7 RNApolymerase in vitro, and then the mixture of PD-1-sgRNAand Cas9 mRNAwas delivered into activated T cells by nucleofection. The efficiency of gene knockout was confirmed by sequencing. The phenotypes of T lymphocytes and the expression of PD-1 after gene knockout were analyzed by Flow cytometry. The proliferation of T lymphocytes was calculated by trypan blue counting. The level of IFN-γ secreted by T lymphocytes was detected by ELISA. Results： ：PD-1-sgRNA and Cas9 mRNA were successfully synthesized in vitro and delivered into T cells by nucleofection. Sequencing technology confirmed that the PD-1 gene sequence was edited and the editing efficiency was 58.3%. The expression of PD-1 on T lymphocyte surface was down-regulated successfully by CRISPR/Cas9 system [(9.6±1.85)% vs (16.2±2.05)%, P<0.05]. The knockout of PD-1 gene did not affect the proliferation and phenotype of T lymphocytes(P<0.05); However, compared with the control group, the level of IFN-γ secreted by T lymphocytes in the PD-1sgRNA group was significantly increased (P<0.01). Conclusion: : CRISPR/Cas9 system can successfully ablate PD-1 gene in human T lymphocytes, which could block the negative regulation of PD-1／PD-L1 and further promote the IFN-γ secretion in T cells.

Objective To summarize the clinical nursing experience for patients with malignant obstructive jaundice after receiving percutaneous biliary radiofrequency ablation (RFA) and stent implantation. Methods The postoperative nursing experience in 9 patients with malignant obstructive jaundice who received percutaneous biliary RFA together with stent implantation were retrospectively analyzed. The postoperative complications and the nursing intervention measures were analyzed and evaluated. Results Biliary RFA and subsequent stent implantation were successfully carried out in all 9 patients. After the procedure, biliary fistula occurred in one patient, biliary hemorrhage in 2 patients and biliary infection in 2 patients. The patient, who developed biliary fistula, died one week later, and the clinical conditions in the remaining 4 patients were improved after symptomatic treatment. During the follow-up period of one month, the patients were in good condition. Conclusion The main purpose of postoperative nursing for patients after receiving percutaneous biliary RFA is to prevent the occurrence of bile duct perforation, hemorrhage, infection, etc. It is very important to keep the patients under close observation and comprehensive nursing so as to make an early detection and timely treatment of such complications, thus to reduce the incidence of complications causing serious consequences as well as to promote an early recovery.

Objective To explore the possible mechanism of miR 34a/Bcl-2 apoptotic pathway in the mouse model of age-related hearing loss.Methods A C57BL/6 mouse model of age-related hearing loss was conducted,and 4-week-old and 12-month-old mice were considered as the objects.The auditory brainstem response (ABR) was used to test the hearing function.The TdT-mediated dUTP nick end labeling (TUNEL) was used to observe the apoptosis of neuron in auditory cortex.The mRNA and protein levels of miR-34a,Bcl-2 and caspase 3 were detected by real-time PCR and Western bloting,respectively.Results The ABR showed that the hearing threshold level at 4,8,16,32 kHz were higher in 12-month-old mice than in 4-week-old mice [(80.0±2.5) dBHL vs.(32.0 ±4.5) dBHL,(74.0±3.5) dBHL vs.(51.0±1.2) dBHL,(86.0±4.6) dBHL vs.(51.0±3.5) dBHL,(87.0±6.6) dBHL vs.(56.0±1.5) dBHL,all P＜0.05].Compared with 4 week-old mice,the total number of neurons in auditory cortex was decreased,the number of apoptosis neurons was increased,the expressions of miR-34a (t=6.02,P=0.001),Bax (t=6.51,P=0.012) and Caspase 3 (P=0.023) rised,and the expression of Bcl-2 (t=7.12,P=0.032) declined in 12 month-old mice.Conclusions The activation of miR-34a/Bcl-2 apoptotic pathway may be one of the mechanisms of age-related hearing loss.

Objective To investigate the roles of cardiac activity index(Tei index) on evaluation the right ventricular function after neonatal asphyxia.Methods Sixty neonatal asphyxia who included 35 cases of mild asphyxia(mild asphyxia group) and 25 cases of severe asphyxia(severe asphyxia group) and 30 cases of normal full-term newborns(control group) were selected.Echocardiographic examinations were performed on 24-48 h after birth,which included pulmonary artery systolic pressure (PASP),right ventricular ejection fraction(RVEF),tricuspid early diastolic peak(peak E) and late diastolic peak(peak A),and E/A ratio was acquired.The right ventricular cardiac activity index (RV-Tei index) was measured by Doppler spectrum.Results There was no significant difference in RVEF,E/A ratio among mild asphyxia group,severe asphyxia group and control group (P ＞ 0.05).RV-Tei index in mild asphyxia group and severe asphyxia group was increased compared with that in control group (0.489 ± 0.090,0.625 ± 0.100 vs.0.345 ± 0.120),and there was significant difference (P＜ 0.05 or ＜0.01).There was significant difference in RV-Tei index between mild asphyxia group and severe asphyxia group (P ＜ 0.05).RV-Tei index in neonatal asphyxia was positively correlated with PASP (r =0.950,P ＜ 0.05),and there was no relationship between RV-Tei index and gestational age,weight,heart rate (r =-0.068,-0.280,-0.360,P ＞0.05).Conclusions Neonatal asphyxia can lead to disorders of the right ventricular function.Tei index can evaluate early overall changes of the right ventricular function and is better than conventional ultrasound technology in neonatal asphyxia.