Objective To investigate the significances of karyotyping analysis on umbilical cord vein blood lymphocytes in the diagnosis of abnormal karyotypes in middle to late period of pregnant fetus.Methods A volume (0.5 ～ 1 ml) of umbilical cord vein blood was extracted from pregnant women in third trimester pregnancy with prenatal detection indications,and collected in sterilized anticoagulant tube.Lymphocytes were cultured and collected for karyotyping analysis after fixed and dropped on slides.Data were analyzed statistically.Results Lymphocytes were cultured successfully in 1 211 cases out of total 1 213 cases collected.Totally 142 abnormal karyotypes were found,which includes 81 cases (detection rate 6.68 ％) of non-heteromorphic abnormal chromosomes and 61 cases (detection rate 5.03％) of heteromorphic chromosomes.Among these abnormal karyotypes,50 cases (accounting for 35.21％ in total abnormal cases) of aneuploidy include 4 cases of chimerical karyotype.Structural abnormalities were found in 31 cases (accounting for 21.83％ in total abnormal cases) samples including 11 cases of translocations,17 cases of inversion and 3 cases of deletion.Conclusions Based on our findings,karyotyping analysis on umbilical cord vein blood lymphocytes could be an effective method for detect abnormal karyotypes in middle to late period of pregnant fetus and played an important role in prenatal diagnosis.

Objective To study the effect of Guiji cream in the treatment of fissure and permeability of skin.Methods The models of fissure Japonic rabbits were established to observe the effects of skin cream on inhibiting fissure.Rabbits back area was divided into 6 blocks and given different medicines in six different groups:GuiJi cream group of low dosage,middle dosage and high dosage (0.05、0.1、0.2 g/m-2 crude drug of Bletilla colloid respectively),urea frost cream group(0.1 g/m-2),normal saline group and control group.Observe the changes of skin surface symptoms every day.The permeability test was performed on 36 KM mouse and randomly divided into control group,GuiJi cream group and urea frost cream group(0.2 g/m-2).Apply drugs to the back and observe the permeability change of cilia capillary,and auricle swelling degree in mice of each group.Results The curative criteria for Guiji groups was 3.8～4.1,the inhibition rate of auricular swelling was 75％.GuiJi cream could inhibit xylene-induced ear edema (P＜0.05) ; the Inhibition rate of vascular permeability was 45.08％,The ear swelling induced by xylene and increased capillary permeability induced by acetic acid in mice were inhibited significantly (P＜0.05) by Guiji cream which had obvious effects on fissure as well as the permeability.Conclusion Guiji chapped skin cream played significant functions in treating fissure and acting anti-inflammatory.

Objective To investigate the relationship between bladder outlet obstruction(BOO) and serum prostatic specific antigen(PSA) in patients with benign prostatic hyperplasia(BPH).Methods The study of pressure-flow urodynamic examination,IPSS score and serum PSA were performed in 253 cases with BPH.Results Based on the bladder outlet obstruction index(BOOI),all patients were divided into three groups: Definite BOO group had 156 patients,mild BOO group had 61 patients,no BOO group had 36 patients.The data of tPSA were(4.54?1.71) ?g/L,(2.45?1.74) ?g/L and(1.85?1.71) ?g/L respectively,there were significant differences between definite BOO group and other two groups(P0.05).Conclusion As a easy detection index,serum PSA may partially reflect the severity of BOO,combined with IPSS score,uroflow, type of B supersonic wave,it would give some guide for patients with BPH to select therapies and judge the prognosis.

Objective To investigate the therapeutic effect of hypertonic saline (HS) and hypertonic saline with taurine (HST) hemorrhagic shock and hemorrhagic shock with hypernatremia rats.Methods Hypernatremic dehydration and hemorrhagic shock models were produced by Trachtma's and Krausz's methods in Sprague-Dawley rats. Hemorrhagic changes of mean arterial blood pressure (MABP), left ventricular end diastolic pressure (LVEDP), ±dp/dtmax and heart rate (HR) were registered on polygraph. Plasma Na+, urea lactate and taurine content were assayed.Results After treatment with HS, the hemodynamic changes of hemorrhagic shock rats were significantly alleviated, and tissue fluid redistributed. When the hemorrhagic shock animals were treated with HS containing taurine (HST), in comparison with HS treatment, the hemodynamic improvement and hemodilution were more obvious. When the hemorrhagic animals complicated with hypernatremia were treated with HS, the symptoms of dehydration and shock further deteriorated, when the hemorrhage with hypernatremia animals were treated with an infusion of HST, the symptoms of dehydration and shock were significantly ameliorated.Conclusion The therapeutic effect of HS with taurine is obviously better than HS alone. So when hemorrhagic shock was complicated with hypernatremia, HS with taurine is recommended in stead of HS alone.

Objective To explore the value of the plasma miR-193a-5p level on diagnosis and monitoring the response to treatment in acute myeloid leukemia (AML). Methods Peripheral blood samples were obtained from AML patients enrolled in hematology department of the Second Hospital of Shanxi Medical University from July 2015 to December 2015, including 30 de novo AML patients, 9 patients in complete remission (CR) and 6 patients in relapse. Peripheral blood samples from 15 healthy people were randomly choosed as the health control group. Plasma miR-193a-5p expression levels were detected by using quantitative real-time reverse transcription polymerase chain reaction (qRT-PCR). Results The plasma miR-193a-5p relative expression level of AML patients group [0.465 6 (0.103 1-5.000 2)] was obviously lower than that of health control group [0.766 1 (0.052 1-3.134 4)] (U= 122, P= 0.018 7). The plasma miR-193a-5p relative expression levels of de novo group and relapse AML group were significantly lower than those of CR group and health control group (P<0.05), and there was no significant difference between the CR group and health control group (U= 56, P= 0.511 9). No significant correlation was found between the plasma miR-193a-5p level and age, gender, blast percentage of the bone marrow, peripheral blood leukocyte count, platelet count, CD34+cells'percentage and so on. Conclusion The decreased plasma miR-193a-5p expression level can be served as a new and noninvasive biomarker for the evaluation of diagnosis and treatment in AML.

Objective:To investigate the serum levels of myonectin, corticostatin and Delta like ligand 4 (DLL4) in patients with type 2 diabetes mellitus (T2DM) diabetes retinopathy (DR) and their clinical significance.Methods:A prospective selection of 341 T2DM patients admitted to Beijing Hospital of Traditional Chinese Medicine, Huairou Hospital from May 2020 to March 2022 was conducted. The patients underwent fundus examination and were divided into a non DR group ( n=85 cases) and a DR group ( n=256 cases) based on DR diagnostic criteria. The DR group was divided into non proliferative and proliferative types according to the staging criteria in China′s DR clinical diagnosis and treatment guidelines, with 142 cases and 114 cases, respectively; 190 healthy individuals who underwent physical examinations in our hospital during the same period were selected as the control group. Enzyme linked immunosorbent assay was used to detect the levels of serum sarconectin, corticostatin, and DLL4 in three groups, collect patient data, and detect biochemical indicators. Logistic regression analysis was used to analyze the influencing factors of DR, and Pearson correlation analysis was used to investigate the relationship between serum sarconectin, corticostatin, DLL4, glucose and lipid metabolism, and insulin resistance; Logistic regression analysis was used to analyze the influencing factors of DR, and Pearson correlation analysis was used to investigate the relationship between serum sarconectin, corticostatin, DLL4, glucose and lipid metabolism, and insulin resistance; The receiver operating characteristic (ROC) curve was used to analyze the diagnostic value of serum sarconectin, corticostatin, and DLL4 in DR. Results:The levels of serum sarconectin and DLL4 in the DR group and non DR group were higher than those in the control group, while the levels of corticostatin were lower than those in the control group (all P<0.05); The levels of sarconectin and DLL4 in the DR group were higher than those in the non DR group, while the levels of corticostatin were lower than those in the non DR group (all P<0.05). The serum levels of sarconectin and DLL4 in proliferative DR patients were higher than those in non proliferative DR patients, while the levels of corticostatin were lower than those in non proliferative DR patients (all P<0.05). The duration of T2DM in the DR group was longer than that in the non DR group, with smoking and alcohol consumption, systolic blood pressure, fasting blood glucose, glycated hemoglobin, triglyceride and insulin resistance index (HOMA-IR) were higher than those in non DR group (all P<0.05). Logistic regression analysis showed that the course of T2DM, systolic blood pressure, smoking and alcohol consumption, glycated hemoglobin, triglycerides, myonectin, corticostatin and DLL4 were the influencing factors of DR (all P<0.05). Pearson correlation analysis results showed that serum sarconectin, DLL4, and fasting blood glucose, glycated hemoglobin, fasting insulin and HOMA-IR were positively correlated (all P<0.05), while cortisol was negatively correlated with fasting blood glucose, glycated hemoglobin and HOMA-IR (all P<0.05). The ROC analysis results showed that the area under the curve (AUC) for the diagnosis of DR was 0.691, 0.745, 0.749, and 0.861 for sarconectin, corticostatin, and DLL4 alone and in combination, respectively. The combined application had higher diagnostic value. Conclusions:Patients with T2DM complicated with DR have elevated levels of serum sarconectin and DLL4, while decreased levels of corticostatin, which are closely related to glucose and lipid metabolism and insulin resistance, and are influencing factors for the occurrence of DR. Combined detection of the three can improve the value of predicting DR.

Objective: To investigate the effect of biology and mTOR pathway activity of down-regulated TSC2 gene expression on U937 leukemia cells. Methods: Gene expression was down-regulated by lentivirus induced RNA interference on TSC2 high expressed U937 cell line; the proliferation, apoptosis and differentiation were detected by CCK-8 assay, colony formation assay and flow cytometry; the gene expression level and protein kinase activity were detected by qRT-PCR and Western blot. Results: Down-regulated expression of TSC2 gene promoted U937 cell proliferation and colony formation ability (P<0.05) . The proportion in G₀/G₁ phase of TSC2 down-regulated U937 cell was much lower than that of the control cells [ (52.53±3.75) % vs (75.10±4.33) %, t=6.829, P=0.002], the S phase [ (22.43±1.00) % vs (15.47±1.20) %, t=-5.581, P=0.019] and G₂/M phase [ (25.03±4.34) % vs (14.33±0.91) %, t=-5.413, P=0.013] was remarkably higher than that of the control cells (P<0.05) . There were no statistically significant differences in cell apoptosis and differentiation (P>0.05) . Down-regulation of TSC2 led to the increased activity of mTOR, 4EBP1 and S6K1, but did not influence the activity of AKT. The expressions of proliferation related cyclinD1, c-myc and PTEN were also up-regulated after TSC2 silenced, but the expressions of P27KIP and BCL-XL were not changed. Conclusion Downregulation of TSC2 could promote the proliferation of U937 cells through up-regulation of mTOR activity.

Objective To investigate the mutations of epigenetic regulation factor ASXL1 gene in myelodysplastic syndrome(MDS).Methods Mutation analysis of ASXL1 gene in 53 de novo MDS patients and 20 healthy persons was performed by using polymerase chain reaction(PCR)followed by sequence analysis at DNA level.The clinical and laboratory characteristics were compared in MDS patients with ASXL1 gene mutation and ASXL1 wild type.ASXL1 mutation in mRNA level was detected by using reverse transcription PCR(RT-PCR)followed by sequence analysis.Results ASXL1 gene mutations were observed in 9 cases(16.9%)of 53 MDS patients.6 mutation types were detected,including 4 frameshift mutations types(2 cases with p.Glu635ArgfsX15,3 cases with p.Gly646TrpfsX12,1 case with p.Ala640GlyfsX14 and 1 case with p.Gly790TrpfsX10)and 2 nonsense mutation types(1 case with p.Gln1063X and 1 case with p.Gln695X).All the mutations were heterozygous,and p.Gly790TrpfsX10 and p.Gln695X were new mutation types.In addition,a single nucletide polymorphism(SNP)p.Gly652Ser was also detected in 4 cases with MDS.5 cases of p.G652S SNP and 1 case of p.Leu1173Leu SNP were detected in 20 healthy people.Frameshift mutation(p.Gly646TrpfsX12)could be detected at mRNA level by using RT-PCR.Differences were not observed in red blood cell counts,white blood cell counts,platelet counts,hemoglobin levels,reticulocyte,neutrophil granulocyte,the peripheral blood lymphocytes percentage,T-cell subsets in the peripheral blood,the proportion of primitive cell in the marrow and MDS types between the patients with ASXL1 gene mutation and ASXL1 wild type patients(P >0.05).Conclusion There is a high frequency of ASXL1 gene mutation in MDS patients,which can be detected at mRNA level.

Objective: To study the distribution of natural killer (NK) cells and the CD38 positive subpopulations in different tissues in rheumatoid arthritis (RA) patients and the mechanism of CD38 agonist. Methods: The levels of NK cells, CD38 positive subpopulation and NK cell surface chemokine receptors (CXCR3, CCR5), as well as granzyme B and perforin were detected by flow cytometry in peripheral blood and knee synovial fluid. The isolated cells were then cultured in vitro and divided into 3 groups, namely blank control group, IB4 stimulation group (Anti-CD38 mAb), IL-2 and IB4 co-stimulation group. And the concentration of tumor necrosis factor-α (TNF-α) and interleukin (IL)-6 in culture supernatants was determined by enzyme linked immunosorbent assay (ELISA). Liposome CD38-siRNA was transfected into peripheral blood lymphocytes of RA patients, and transfection efficiency was detected by reverse transcription-polymerase chain reaction (RT-PCR). The effect of CD38 and CD38-siRNA silencing on the expression of phospholipase C-gamma 1 (PLC-γ1) protein in the peripheral blood lymphocytes of RA was detected by Western blot. Results: The expression of CCR5 and CXCR3 in NK cells in peripercal NK cells of RA was significantly higher than that in normal healthy subjects (P<0.05). The two chemokine receptors (CXCR3, CCR5) were mainly expressed in CD38⁺ NK cell subsets. The levels of granzyme B and perforin in synovial lymphocytes cells were significantly higher than those in peripheral blood lymphocytes (P<0.05). The secretion of IL-6 and TNF-α in synovial and peripheral blood lymphocytes cells existed significant difference only in IL-2 and IB4 co-stimulation group (P<0.05). The levels of PLC-γ1 protein in the peripheral blood lymphocytes of RA patients was significantly decreased than that in the blank control group after the treatment with CD38-siRNA (P<0.05). Conclusions The synovial NK cells are more lethal than the peripheral NK cells in the RA patients. CD38 might mediate the phosphorylation of intracellular proteins via the Protain kinase C (PKC) pathway. Blocking CD38 molecules can reduce the phosphorylation level of intracellular proteins. CD38 maybe involved in the mechanism of chemotaxis and killing capability of NK cells.