Objective To observe the changes of serum anti-mttllerian hormone (AMH) level in women with premature ovarian fail-ure (POF) and its clinical significance. Methods Fasting serum levels of AMH in 25 POF patients were measured with enzyme-linked im-mtmosorbent assay (ELISA). Twenty healthy women matched for age and body mass index (BMI) were chosen as controls. Results The serum AMH level in POF group was (1.1±0.1) pmol/L, which was significantly lower than that of control group(P < 0.01). The serum AMH levels in 20 patients of POF group were undetectable. The other 5 patients were treated with artificial cycle at first, and then ovulated with hMG/HCG.2 patients had ovulation and 1 patient was pregnant. Conclusion The serum AMH level in POF patients was significantly decreased, and the measure for serum AMH had some significance for POF diagnosis and treatment.

Objective To determine the role of interleukin-18(IL-18) and vascular endothelial growth factor (VEGF) in the ovari-an hyperstimulation syndrome (OHSS) development. Methods Serum and follicular IL-18 and VEGF levels were measured with ELISA in 66 IVF patients, who were divided into three study groups according to OHSS risk factors on the day of ovum retrieval and whether will devel-op OHSS in future or not. 20 patients had no OHSS risk factors (normal group), 28 patients had OHSS risk factors without developing OHSS (OHSS risk group) and 18 patients had OHSS risk factors and developing OHSS in future (OHSS group). Results Serum and follicular IL-18 levels in OHSS group were significantly higher than those of other two groups(P<0.01). Serum and follicular VEGF levels in OHSS group were also significantly higher than those of other two groups(P<0.01). Conclusion Serum and follicular IL-18 and VEGF levels in OHSS group increased significantly, which suggested IL-18 and VEGF were positive correlated with OHSS development. And follicular IL-18 and VEGF levels in serum on the day of ovum retrieval have a prediction role in OHSS development.

AIM:To discuss the effect of Shenmai injection on insulin resistance ( IR) in 3T3-L1 cells and its mechanisms.METHODS:3T3-L1 preadipocytes were induced by chemical reagents to differentiate into fully differentiated adipocytes.Oil red O staining was used to detect the differentiation level of the adipocytes .The insulin-resistant 3T3-L1 cell model was demonstrated using insulin , which was confirmed by glucose concentration in cell supernatant .The IR cell model was given 10 μmol/L rosiglitazone , 25 and 50 g/L Shenmai injection and normal saline for comparison .MTT assay was used to assess the cell activity of 3T3-L1 cells which was treated with drugs for 8, 16, 24 and 36 h.Glucose oxidase method was used to detect the glucose concentration in the cell supernatant at 8, 16 and 24 h.The protein levels of glucose transporter-4 (GLUT4), phosphatidylinositol 3-kinase (PI3K), AKT and p-AKT were determined by Western blot .RE-SULTS:3T3-L1 adipocytes were successfully induced as shown by the positive oil red O staining .The IR cell model was demonstrated , and glucose concentration in the cell supernatant after treatment with Shenmai injection showed that Shenmai injection reduced the IR in 3T3-L1 cell model.The protein levels of GLUT4, PI3K and p-AKT increased compared to con-trol group.CONCLUSION:Shenmai injection reduces the IR in 3T3-L1 cell model, which functions by increasing the protein levels of GLUT4, PI3K and p-AKT.

Objective To examine the relationship between the concentration of 25-hydroxy vitamin D3 [25-(OH) D3] in the serum and the body mass,the severity of obesity,body mass index(BMI),blood lipid,and their predicting role in obesity children.Methods The study recruited 244 subjects,who see the doctor in Wuxi Maternal and Child Health Hospital,Childhood Nutrition Outpatient from Jul.2011 to Feb.2013.The intake dose of vitamin D each day was investigated,and weight,height,BMI,concentration of 25-(OH) D3 in serum,and microelement were also measured.In addition,lipid metabolism of 38 cases with obesity over 3 years old was determined.Results 1.The serum 25-(OH) D3 concentration of obese children was (68.31 ± 23.06) nmol/L.The concentration of 25-(OH) D3 was lowest in the group of obese children over 36 months of age[(55.03 ± 15.18) nmol/L].2.The concentration of 25-(OH) D3 in the group of obese and overweight children was far lower than that of the children in the normal group (F =4.739,P ＜0.05).3.The concentrations of 25-(OH) D3 in the severely obese children was significantly lower than that of the mild and moderate obesity children(F =9.711,P ＜ 0.05).4.There were significantly inverse associations of serum 25-(OH) D3 with weight,weight and height percentage,BMI (r =-0.365,-0.237,-0.175,all P ＜ 0.001).5.There were significantly inverse associations between the concentration of 25-(OH) D3 in serum with weight,triglyceride in obese children more than 3 years old (r =0.476,-0.324,all P ＜ 0.05).Conclusions The decreasing level of 25-hydroxy vitamin D3 in the serum was associated with obesity.The cause of it might be the increase of the obese adipose tissue,vitamin D getting trapped in fat cells,and all these factors can lead to a less serum vitamin D levels.The vitamin D consumption of obese children is higher than that of normal children,and should supply more vitamin D to reach normal 25-(OH) D3 level.

Objective Study on the absorption,distribution,transformation,excretion and toxicity of four forms of arsenic in rats.Methods Four thousand and fifty data were obtained from 28 days animal metabolism experiments of 81 rats exposed to the four As species.Distribution,metabolism,excretion,and subacute toxicity of 4 As species were compared by analyzing the arsenate [As(V)],arsenite [As(Ⅲ)],methyl arsenate (MMAV) and dimethyl arsenate (DMAV) content of rat feces (excrement and urine),blood,seven organs,and liver and kidney pathology.Results After oral administration of each As species,82.9％ of As(Ⅲ),85.1％ of As(V),95.0％ of MMAV and 96.2％ of DMAV were accumulatively secreted via feces and urine,while 16.2％ of As(Ⅲ),14.1％ of As(V),4.65％ of MMAV and 0.120％ of DMAV were detected in blood.The DMAV accumulated in blood and urine after dosing As(Ⅲ),As(V) and MMAV,and the content of DMAV in the blood was 8 times greater than urine.Four kinds of arsenic had effects on rat liver and kidney function,the influence on liver was acute phase effect while that on renal was cumulative effects.The effect of As(Ⅲ) was the most significant,followed by MMAV.Four kinds of arsenic could cause tissue deformation and inflammatory cells infiltration,but showed no difference between groups.Conclusion The distribution,metabolism,excretion,and toxicity of the four kinds of arsenic in rats were different in varying degrees.The research of health risk for arsenic should consider its different forms.

OBJECTIVE: To investigate the effect of 5-Aza-dC and TSA to tumor suppressor gene RASSF1A expression and methylation level in Hep-2 cell line. METHOD: Hep-2 cell line were cultured in vitro and handled with 5-Aza-dC and TSA. Detected RASSF1A expression and methylation level were detected before and after drug intervention using Realtime PCR and MSP. RESULT: (1) Before intervention with drug, tumor suppressor gene RASSF1A was weakly expressed and methylated in Hep-2 cell line. (2) With the effect of 5-Aza-dC and TSA, the methylation of RASSF1A gene was reversed. And the effect of combination of 5-Aza-dC and TSA was similar with 5-Aza-dC alone. There was no obvious effect using TSA alone. (3) With the effect of 5-Aza-dC and TSA, the expression of RASSF1A was improved. And the effect of 5-Aza-dC was stronger than TSA. Synergetic effect was found when using 5-Aza-dC and TSA simultaneously. CONCLUSION In Hep-2 cell line, Promoter methylation of tumor suppressor RASSF1A may play a very important role in loss of gene expression, but it is not the only cause. 5-Aza-dC and TSA can improve RASSF1A expression by reversing DNA methylation and histone deacetylation.
Antimetabolites, Antineoplastic ; pharmacology ; Azacitidine ; analogs & derivatives ; pharmacology ; Cell Line, Tumor ; DNA Methylation ; Gene Expression ; Gene Silencing ; Genes, Tumor Suppressor ; Humans ; Hydroxamic Acids ; pharmacology ; Promoter Regions, Genetic ; Tumor Suppressor Proteins ; genetics

Objective:To explore the performance characteristics of the digital clock-drawing test(dCDT) for amnestic mild cognitive impairment(aMCI), and its diagnostic value for aMCI patients compared with the traditional clock-drawing test (tCDT).Methods:Total 81 middle-aged and elderly outpatients in Affiliated Hospital to Shanxi Medical University from November 2020 to May 2021 were selected, including 42 cognitively normal people (control group) and 39 aMCI patients (aMCI group). The dCDT developed by our team was used to collect drawing process parameters (such as stroke length, time and speed). The Cognitive Domain Indexs of Montreal Cognitive Assessment (MoCA) were calculated using the CDIS scoring method, and the correlation between dCDT parameters and MoCA indexs were analyzed.Logistic regression analysis was used to construct the predictive model, and the sensitivity and specificity of different methods for the diagnosis of aMCI patients were compared by the area under the ROC curve.Results:(1) The total time(51.25(38.80, 63.75)s vs 42.42(33.64, 51.91)s) and time in air(36.34(26.81, 47.25)s vs 28.47(22.37, 33.98)s) of the aMCI group were significantly higher than those of the control group, and the minute hand/hour hand ratio(1.23±0.35 vs 1.39±0.34), strokes per minute((31.31±10.44) vs (41.05±9.48))and tCDT score(3.0(3.0, 4.0), 4.0(3.0, 4.0))were significantly lower than those of the control group, and the differences were statistically significant (all P<0.05). Other dCDT parameters were not statistically significant between the two groups ( Z=-1.835--0.440, P>0.05). (2) Correlation analysis showed that the total time was negatively correlated with MoCA MIS( r=-0.224, P=0.049), LIS( r=-0.237, P=0.037)and AIS( r=-0.236, P=0.038); time in air was negatively correlated with MoCA MIS( r=-0.268, P=0.018), LIS( r=-0.271, P=0.016), AIS( r=-0.259, P=0.022)and OISA( r=-0.267, P=0.018); the minute hand/hour hand ratio was positively correlated with MoCA EIS( r=0.259, P=0.022)and VIS( r=0.309, P=0.006); the strokes per minute was positively correlated with MoCA MIS( r=0.376, P=0.001), EIS( r=0.290, P=0.010), VIS( r=0.294, P=0.009), AIS( r=0.238, P=0.036)and OISA( r=0.301, P=0.007). (3)dCDT model composed of the pre-second hand latency, the ratio of minute hand/hour hand, and the strokes per minute can correctly classify 77.8% of aMCI, with a sensitivity of 74.36% and a specificity of 80.95%.Its diagnostic power for aMCI was significantly higher than the tCDT scoring( Z=2.335, P=0.02). Conclusion:The cognitive impairment in aMCI can be detected by dCDT, and different dCDT parameters can reflect the impairment of different cognitive domains.Compared with tCDT scoring, dCDT can improve the diagnostic efficacy of aMCI patients.

OBJECTIVETo construct dengue virus-specific full-length fully human antibody libraries using mammalian cell surface display technique.

METHODSTotal RNA was extracted from peripheral blood mononuclear cells (PBMCs) from convalescent patients with dengue fever. The reservoirs of the light chain and heavy chain variable regions (LCκ and VH) of the antibody genes were amplified by RT-PCR and inserted into the vector pDGB-HC-TM separately to construct the light chain and heavy chain libraries. The library DNAs were transfected into CHO cells and the expression of full-length fully human antibodies on the surface of CHO cells was analyzed by flow cytometry.

RESULTSUsing 1.2 µg of the total RNA isolated from the PBMCs as the template, the LCκ and VH were amplified and the full-length fully human antibody mammalian display libraries were constructed. The kappa light chain gene library had a size of 1.45×10(4) and the heavy chain gene library had a size of 1.8×10(5). Sequence analysis showed that 8 out of the 10 light chain clones and 7 out of the 10 heavy chain clones randomly picked up from the constructed libraries contained correct open reading frames. FACS analysis demonstrated that all the 15 clones with correct open reading frames expressed full-length antibodies, which could be detected on CHO cell surfaces. After co-transfection of the heavy chain and light chain gene libraries into CHO cells, the expression of full-length antibodies on CHO cell surfaces could be detected by FACS analysis with an expressible diversity of the antibody library reaching 1.46×10(9) [(1.45×10(4)×80%)×(1.8×10(5)×70%)].

CONCLUSIONUsing 1.2 µg of total RNA as template, the LCκ and VH full-length fully human antibody libraries against dengue virus have been successfully constructed with an expressible diversity of 10(9).

Animals ; Antibodies, Viral ; CHO Cells ; Cell Surface Display Techniques ; Cricetinae ; Cricetulus ; Dengue Virus ; immunology ; Gene Library ; Genetic Vectors ; Humans ; Immunoglobulin Heavy Chains ; immunology ; Transfection

Human coronavirus OC43 (HCoV-OC43) and severe acute respiratory syndrome coronavirus 2(SARS-CoV-2) belong to the β-coronavirus genus. Since the discovery in 1967, HCoV-OC43 has been continuously circulating in human population and has become one of the common seasonal respiratory viruses. SARS-CoV-2, which has a higher morbidity and fatality rate, appeared at the end of 2019, followed by the emergence of a variety of variants, and the transmission and infection capacity of SARS-CoV-2 has been enhanced. HCoV-OC43 may be similar to SARS-CoV-2 in terms of genomic structure and function, species evolution, epidemic characteristics and clinical manifestations. In this review, the epidemiology, genomics, phylogenetic evolution and other aspects of HCoV-OC43 and SARS-CoV-2 were analyzed. Such an analysis would be helpful to understand the association and differences between the two viruses, and provide reference for understanding the potential threats of HCoV-OC43.

Since an epidemic occur of Corona Virus Disease 2019(COVID-19) in December, 2019, all the dental healthcare service providers in our country have been greatly impacted. The strategy of managing the dental clinics is quite different from that of the medical healthcare clinics, and the key point of the administration of those dental healthcare providers is to focus on the management of outpatient care because they only supply just a little bit of inpatient care service but quite an large amount of outpatient care service. So we think the next step is to make plausible and effective scenarios to protect our dental healthcare staff and patients against corona virus infection during the treatments procedures after the reopening all of our dental clinics. To overcome this harsh condition, the infection prevention and control strategies adopted by the Stomatological Hospital, Southern Medical University were designed to be flexible and could be adjusted promptly according to the national and local governmental orders and latest guidelines released by the Centers for Disease Control and Prevention. All these prevention procedures and protocols were customized to fit our own situation and have been updated for several times based on the latest global pandemic reports. After going through the hardest time in the past four months, it’s considered that our COVID-19 prevention rules have been proved to be efficient and work well. Further more, it has made massive progress for the hospital in improving the capability of dealing with this state of emergency, especially by previewing and triaging patients strictly to cut off the possible coronavirus spreading from the original step, enhancing the standard precautions and those specific protocols made for minimizing the droplets, aerosol and contact transmission of COVID-19 indoors. Besides, a daily supervision system was set up as a routine job and a team of qualified infection prevention specialists were assigned to check and report every incorrect details during the whole procedure of dental practice. Meanwhile, the safety and well-being of the public and our medical workers could be also guaranteed through following those detailed prevention scenarios.