Objective: To investigate the expression of MDR1 and GFP in the human hematopoietic cells mediated by adeno-associated virus. Methods: The GFP gene was transferred into the human hematopoietic cells by AAV vectors and created strong visible fluorescence by purely molecular biological means. Using adeno-associated virus vectors, we have transferred human mdr-1 gene into human hematopoietic cells and investigated the drug resistence of human hematopoietic cells modified with mdr-1 gene. PCR analysis confirmed that mdrl cDNA had been successfully transferred into the human hematopoietic cells. An assay of MTT proved that the human hematopoietic cells modified by mdrl gene had resistance to colchicine. Results: It was about 30% of the hematopoietic cells that expressed the green fluorescent proteins. The resistance of hematopoietic cells was increased parently when the cells were infected by the crude virus stocks. Conclusion: It is conducted that the AAV vector could successfully transfer the foreign gene into the human hematopoietic cells. The cells modified with mdrl gene have increased the resistance to drugs.

Objective:To investigate the onset mechanism of cervical vertigo from the proprioceptive sensation and provide clinical basis for its treatment.Methods:Among the 121 cases that conformed to the diagnostic criteria of cervical vertigo,70 cases who presented with negative neck-rotation test and cervical vertigo without obvious vascular factors by transcranial Doppler(TCD)were assigned to the observation group,while 51 cases who presented with positive neck-rotation test and cervical vertigo due to spasm of vertebral basal artery or insufficient blood supply by TCD were assigned to the control group.The cases in the two groups were treated once every day,5 days make up one treatment course and the resuIts were statistically analyrzed after one treatment course.Results:The skull triaxial spatial offset of the cases in the two groups were significantly reduced after the treatment (P<＜0.01).However, there was no statistically significant difference between the skull triaxial spatial offset between the two groups (p＞0.05).It is not conclusive that the therapeutic effect in the two groups was significantly different after one treatment course.Conclusion:Tuina manipulation therapy can improve the skull spatial ofrset repositioning ability of the Patients.

Objective: To measure serum monocyte chemoattractant protein-1 (MCP-1) levels and study its associations with lipoproteins in patients with polycystic ovary syndrome (PCOS). Methods: Sixty-five PCOS women and 20 ovulating normal women with body mass index (BMI)

Objective:To investigate the impact of adding human menopausal gonadotropin (hMG) for in vitro fertilization-embryo transfer pregnancy outcomes in a standard population of non-advanced age with normal ovarian reserve function using a long follicular phase protocol.Methods:Clinical data of 489 patients with normal ovarian reserve function, who were admitted from January 2018 to January 2020 in the Affiliated Hospital of Guizhou Medical University and underwent in vitro fertilization for the first time with the long follicular phase protocol in fresh cycles, were retrospectively analyzed. The patients were divided into three groups according to whether or not to add urine-derived hMG and the timing of addition: non-addition group (group A), medium-term hMG group (group B1), whole course hMG group (group B2); the laboratory parameters of each group were observed, and the effect of ovulation induction drugs and pregnancy outcomes were compared.Results:The ages of B1 and B2 groups were significantly higher than that of group A ( P=0.019 and P=0.011). The basal FSH level of group B2 was significantly higher than those of group A and group B1 ( P<0.01 and P=0.006), and the basal FSH/LH ratio of group B2 was significantly higher than that of group B1 ( P=0.009). Antral follicle counts of group A and group B1 were significantly higher than that of group B2 ( P=0.007 and P=0.017). The superior embryo rate of group B2 [(47±27)%] was significantly higher than that of group A ( P=0.017). The embryo implantation rate of group B1 was significantly lower than those of group A and group B2 ( P=0.043 and P<0.01). The clinical pregnancy rate of group B2 [76.7% (155/202)] was significantly higher than those of group A ( P=0.039) and group B1 ( P<0.01). The live-birth rate of group B2 [67.3% (136/202)] was significantly higher than those of group A ( P=0.017) and group B1 ( P=0.001). Conclusions:For non-advanced aged patients with normal ovarian reserve function, the long protocol of follicular phase is suitable for those with relatively low ovarian reserve function. Adding hMG in the whole course of ovulation induction after gonadotropin-releasing hormone agonist reduction could improve the pregnancy outcomes by improving the quality of embryos.

Objective To construct the eukaryotic expression vector harboring the fragment of Alia gene, and to investigate the effects of it on the signal of quorum sensing and virulence factors producted by Pseudomonas aeruginosa(Pa). Methods The plasmid pET-AiiA was cutted by Nhe Ⅰ and Xho Ⅰ , then the AiiA fragment was cloned into eukaryotic expression vector pEGFP-N2. After the plasmid was transfected into A549 cells, the protein was extracted and AiiA protein was found in it by Western blot. After the extrac- tion was admixed into the LB broth, from culture supernatant extracts of Pa, the N-acylhomoserine lactone (AHL) was detected by bioassay, and the expression of pyocyanin and elastase were assayed by RT-PCR and optical density. Results The fragment of AiiA gene was cutted and then cloned into pEGFP-N2. AiiA protein was found in the transfected cells. After admixed with the extract harboring AiiA protein, in Pa medium, the AHL was hydrolyzed, and the expression of pyocyanin and elastase were reduced. Conclusion The virulence factors synthesized by Pa were reduced by the AiiA protein expressed in eukaryotic cell.

Objective To study the spatial-temporal dynamical features of measles in Zhejiang province.Methods Data was from the China Disease Surveillance Information System and China Immunization Program Information Management System.Power-law method on spatial-temporal-multicomponent model was used to analyze the epidemic characteristics of measles in the districts of Zhejiang province.Results The incidence of measles in Zhejiang province was 2.72/ 100 000 (1 494 cases) in 2013.Compared to the first order adjacent matrix,Power-law method showed a lower value of Akaike information criterion.The follow-up impact from the previous measles epidemic was strong to the Keqiao,Xiaoshan and Yuecheng districts with the autoregressive component as 1.39,0.88 and 0.77,respectively.Local risk of measles seemed high in Keqiao,Qujiang and Xiaoshan districts with the endemic component as 4.06,3.74 and 3.55,respectively.Impact of the epidemic to the nearby districts was large in Keqiao,Shangyu districts and Jiande city with epidemic components as 3.08,2.54 and 2.21,respectively.Conclusion The spatial-temporal feature of measles in several districts of Zhejiang province appeared heterogeneous,suggesting the specific strategies should be taken to control the epidemics of measles.

Objective:To study the level and deficiency of 25-hydroxy vitamin D [25-(OH)D] in 7215 pregnant women in Dalian district, and provide a scientific evidence for the prevention of vitamin D deficiency and reasonable supplementation.Methods:During January to December of 2018, the basic information and blood samples of 7215 pregnant were collected in Maternal and Child Health Care hospital of Dalian. The level of serum 25-hydroxy vitamin D was detected by chemiluminescence method. All data was analyzed by SPSS 17.0 software.Results:In Dalian district, the average level of serum 25-(OH)D in 7215 pregnant women was 22.74(16.02, 28.62) ng/ml, and the deficiency rate of vitamin D was 33.89%, and the severe deficiency rate was 6.68%. The average level of serum 25-(OH)D in pregnant women aged 18-24 years old, 25-34 years old and 35-45 years old were 20.22(13.61, 25.57) ng/ml, 22.75(16.12, 28.42) ng/ml and 23.60(16.76, 29.92)ng/ml, respectively. With the increasing of pregnant age, the level and the deficiency rate of serum 25-(OH)D was gradually rising with significant difference( P<0.01), and the highest deficiency rate was pregnant aged 18~24 years old with 42.40%. Conclusion:The level of vitamin D of pregnant women in Dalian district was insufficient status, especially for young pregnant women. Health education for vitamin D deficiency should be focused on young pregnant women.

BACKGROUND:Whether activating transcription factor 7 interacting protein(Atf7ip)is involved in the regulation in osteogenic differentiation is still controversial,and studying its impact on osteogenic differentiation and its specific mechanisms is of great significance. OBJECTIVE:To investigate the effect of Atf7ip on bone morphogenetic protein 2 promoting osteogenic differentiation of mouse embryonic osteoblast precursor cells(MC3T3-E1). METHODS:MC3T3-E1 cells cultured in vitro were divided into three groups:normal group,interference group(NC-siRNA group,Atf7ip-siRNA group),and high expression group(CMV-VC group and CMV-Atf7ip group),and were transfected for 24 hours,and then treated with 200 ng/mL bone morphogenetic protein 2 for 0,12,24,and 48 hours,respectively.qRT-PCR was used to detect the mRNA expression levels of Atf7ip,alkaline phosphatase,osteocalcin,type I collagen α1 in the cells of each group.Western blot assay was used to detect the protein expression of osteogenic differentiation markers Sp7 and Runx2,and the expression of Atf7ip binding molecule SETDB1,histone H3 and H3K9me3.Alkaline phosphatase activity was detected by alkaline phosphatase staining. RESULTS AND CONCLUSION:(1)With the increase of bone morphogenetic protein 2 treatment time,the protein and mRNA expression of Atf7ip decreased,while the protein expression of Sp7,Runx2 and the mRNA expression of osteocalcin and alkaline phosphatase increased significantly(P<0.05).There was no significant change in the protein expression of Atf7ip binding molecule SETDB1.(2)Compared with the NC-siRNA group,the protein expression of Sp7,Runx2 and the mRNA expression of osteocalcin and type I collagen α1 were significantly up-regulated(P<0.05),and alkaline phosphatase activity was significantly enhanced;and H3K9 methylation significantly decreased in the Atf7ip-siRNA group(P<0.05).(3)Compared with the CMV-VC group,the protein expression of Sp7 and Runx,as well as mRNA expression of osteocalcin,alkaline phosphatase,and type I collagen α1 was significantly downregulated(P<0.05),and the alkaline phosphatase activity was significantly reduced in the CMV-Atf7ip group,while the H3K9 methylation protein in the CMV-Atf7ip group was significantly upregulated compared to the control group(P<0.05).(4)In conclusion,Atf7ip expression was decreased during bone morphogenetic protein 2-induced osteogenic differentiation of MC3T3-E1,and osteogenic differentiation was significantly increased after knockdown of Atf7ip.Overexpression of Atf7ip significantly weakened osteogenic differentiation,indicating that Atf7ip is a negative regulatory factor of bone morphogenetic protein 2 promoting osteogenic differentiation of MC3T3-E1 cells.

Objective To explore the diagnostic value of transcranial substantia nigra ultrasound in Parkinson's disease(PD)and the clinical characteristics of patients with different substantia nigra(SN)echoes.Methods Sixty patients with PD admitted to Yichun Hospital of Traditional Chinese Medicine from October 2020 to January 2023 were included in study group,and another 60 normal persons who received health examination in our hospital during the same period were included in control group.Both groups were examined by transcranial substantia nigra ultrasound.The SN high-echo area and ratio of total area of hyperechoic substantia nigra to total area of mesencephalon(S/M)of two groups were compared,and the positive rate of PD diagnosed by transcranial substantia nigra ultrasound was calculated.According to the intensity of SN echo,PD patients were divided into PD-SN-group and PD-SN+ group,and the clinical characteristics of patients with different SN echo were compared.Results The SN high-echo area and S/M of study group were significantly higher than those of control group(P<0.05).Transcranial substantia nigra ultrasound showed PD-SN-in 10 cases and PD-SN+ in 50 cases.The positive rate of PD diagnosed by transcranial substantia nigra ultrasound was 83.33%(50/60).There were statistically significant differences between PD-SN+ group and PD-SN-group in gender,age,H-Y stage,age of onset,score of mini mental status examination and score of Montreal cognitive assessment(P<0.05).Conclusion Transcranial substantia nigra ultrasound can better help clinical screening and detection of PD,and improve the positive detection rate of PD.