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Objective:To explore the effectiveness of folate-coupled quantum dots (FA-QD) immunomagnetic beads method for detecting circulating tumor cells (CTC) in epithelial ovarian cancer and the association of CTC with clinicopathological features of tumor patients.Methods:A total of 67 ovarian cancer patients in Shanxi Provincial People's Hospital from August 2019 to January 2020 were selected. Ovarian cancer SKOV-3 cells were divided into 5 cell number gradients (0, 100, 150, 200, 300), the detection rates of CTC were compared by using epithelial cell adhesion molecule (EpCAM) immunomagnetic beads (single standard method) and FA-QD immunomagnetic beads method (double standard method). The number of CTC in peripheral blood of ovarian cancer patients was detected by using FA-QD immunomagnetic beads method, and those with positive CTC under fluorescence microscope were treated as CTC positive patients. The association of CTC with clinicopathological factors and tumor markers of tumor patients was analyzed.Results:The average capture efficiency rate of CTC in SKOV-3 cells detected by FA-QD immunomagnetic beads method (83.4%) was higher than that by EpCAM immunomagnetic beads method (70.3%). Among 67 patients of ovarian cancer, the proportion of CTC positive patients was 30.0% (3/10) in stage Ⅰ-Ⅱ, 91.9% (34/37) in stage Ⅲ, 95.0% (19/20) in stage Ⅳ, and the difference was statistically significant ( P < 0.05). The proportion of CTC positive patients with lymph node metastasis was higher than that of patients without lymph node metastasis [97.1% (33/34) vs. 69.7% (23/33)], and the difference was statistically significant ( P < 0.05). The proportion of CTC positive patients with human epididymis protein 4 (HE4)>110 pmol/L was lower than that of patients with HE4 ≤ 110 pmol/L [58.8% (10/17) vs. 92.0% (46/50)], and the difference was statistically significant ( P = 0.005). There were no statistically significant differences in the proportion of CTC positive patients stratified by age, menopause, pathological differentiation, distant metastasis, carbohydrate antigen (CA) 125, CA199, carcino-embryonic antigen (CEA) (all P > 0.05). Conclusions:FA-QD immunomagnetic beads method can effectively detect CTC in peripheral blood of patients with epithelial ovarian cancer. The level of CTC in patients with epithelial ovarian cancer is related to lymph node metastasis, clinical TNM stage and HE4 level.
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Objective:To observe the clinical characteristics and reproductive outcome of patients with globozoospermia syndrome (also called round-headed spermatozoa).Methods:The data of 5 patients with round-headed spermatozoa and 20 healthy men in the reproductive center of Shanxi Maternal and Child Hospital and Shanxi People′s Hospital from July 2016 to March 2020 were collected. Among them, the wife was healthy. The peripheral blood chromosome, AZF gene, semen routine and morphology, sex hormone series were detected for the man, and the outcome of intracytoplasmic sperm injection (ICSI) was observed for the patients.Results:The average length of infertility in 5 patients with round-headed sperm was (4.4±1.8)years, and the round-headed sperm rate of 5 patients were all over 95% (including 1 case with round-headed sperm rate of 99%, 3 cases with round-headed sperm rate of 98%, and 1 case with round-headed sperm rate of 96%). The chromosome, AZF gene of 5 patients were normal. The sperm motility [progressive motility(PR) rate+ non progressive motility (NP) rate] of patients with globozoospermia syndrome was significantly lower than that of the healthy control group ( P<0.05). There was no significant difference in sex hormone, sperm count, sperm density, PR(%) and sperm DNA fragmentation rate compared with normal fertility group ( P>0.05). All the 5 patients were treated by assisted reproductive technology with ICSI. Two patients were pregnant, including one patient with abortion and one patient with singleton. Conclusions:There are no other abnormal phenotypes in patients with round-headed spermatozoa except for the low morphology and motility of round headed spermatozoa. Assisted reproductive technology may be an effective way to assist pregnancy.
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Objective:To investigate the changes and clinical significance of gastrin 17 (G-17) in patients with diabetic nephropathy (DN).Methods:One hundred and twenty-four DN patients admitted to Hefei Second People′s Hospital from July 2018 to December 2020 were selected as the DN group, and divided into Ⅰ-Ⅱstage subgroup (68 cases) and Ⅲ-Ⅴ stage subgroup (56 cases) according to the stage of DN.Inaddition, 100 cases of type 2 diabetes mellitus(T2DM) patients without DN were selected as the T2DM group, and 100 healthy subjects who examined during the same period were selected as the control group. The levels of G-17, serum creatinine (SCr), evaluated glomerular filtration rate (eGFR) and other index in each group were detected. The normal level of G-17 was 1-7 pmol/L. G-17>7 pmol/L and ≤ 15 pmol/L was as marginal rising, and G-17>15 pmol/L was as rising.Results:The marginal rising rate of G-17 in the DN group was higher than that in the T2DM group: 43.5%(54/124) vs. 23.0%(23/100); the rising rate of G-17 in the DN group was higher than that in the T2DM group and the control group: 21.0%(26/124) vs. 7.0%(7/100), 4.0%(4/100), and the differences were statistically significant ( P<0.05). The marginal rising rate and rising rate of G-17 in Ⅲ-Ⅴstage subgroup were both higher than those in the Ⅰ-Ⅱ stage subgroup and the T2DM group: 58.9%(33/56) vs. 30.9%(21/68), 23.0%(23/100); 32.1%(18/56) vs. 11.8%(8/68), 7.0%(7/100), and the differences were statistically significant ( P<0.05). The marginal rising rate and rising rate of G-17 in DN patients with a disease course of ≥3 years was higher than that in patients with a disease course of <3 years and the T2DM group: 53.0%(44/83) vs. 24.4%(10/41), 23.0%(23/100); 27.7%(23/83) vs. 7.3%(3/41), 7.0%(7/100), and the differences were statistically significant ( P<0.05). Correlation analysis showed that G-17 was positively correlated with SCr ( r = 0.367, P<0.001) and negatively correlated with eGFR ( r = -0.619, P<0.001) in DN patients. Conclusions:The level of G-17 in ND patients is significantly increased, which is closely related to DN staging and can provide an auxiliary indicator for screening renal function in patients with T2DM.
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Objective:To observe the diagnosis effect about the MRI and CT examination for the patients with lacunar cerebral infarction(LAC).Methods: 82 cases early LAC patients during April 2014- June 2016 were diagnosed by two methods, MRI and CT, respectively. And the application effect of the two methods were compared based on image result.Results: In 82 cases of LAC 742 lesions confirmed by MRI and only 145 lesions confirmed by CT, and there was statistical significant between the two methods; especially in front lobe and thalamic, lesions detection rate using MRI was higher than CT, and there was statistical significant between the two methods(x2=6.59,x2=5.64,x2=6.42;P0.05);Conclusion: Both of CT and MRI can be used in early diagnosis of LAC, and MRI examination is more accurate for early or micro lesions and lesions happened in the frontal lobe, thalamus, capsula internal than CT. Therefore, MRI can be used as first choice eximination method in early diagnosis of LAC.
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Objective To establish the rheumatoid arthritis (RA) mouse model induced by glucose-6-phosphate isomerase (GPI),and explore the mechanism of GPI in RA.Methods Totally 36 DBA/1 mice were randomly divided into three groups:test group (injection GPI),positive group (injection of bovine collagen Ⅱ),and negative group (saline).The rates and changes of weight were observed.The score of the arthritis,the ankle histopathological changes and serum GPI content were detected.Results Toes swollen slightly,joint swelling,deformity and accompanied by block were appeared at 35th day in the test group.Compared to the control group,the rates and changes of weight in test group showed a significant difference (P ＜ 0.05).The score of arthritis was showed by x ± s.Compared to the negative group,the test group and positive group were showed significant difference (P ＜ 0.05).A lot of lower synovial lining exudate macrophages,fibroblasts,and other inflammatory cells were increased in the test group.The GPI content in the test group [(0.39 ±0.11)μg/ml] was significantly higher than the negative group [(0.10± 0.06) μg/ml,P ＜ 0.05].Conclusions GPI could induce rheumatoid arthritis in mice.It provides the experimental basis to diagnose RA.
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Objective To obtain the Cyclin D1 through cloning and prokaryotic expression of Cyclin D 1 gene.Methods The total RNA was extracted from liver cancer tissue .The Cyclin D1 cDNA was obtained by reverse transcriptase polymerase chain reaction (RT-PCR).The Cyclin D1 cDNA was sequenced, and sub-cloned to the PET32a+.The prokaryotic expressed was used to obtain the Cyclin D1.Results The 483 bp Cyclin D1 cDNA was obtained.The sequence of Cyclin D1 was corrected.The 36 KD CyclinD1 was obtained by prokaryotic expression .Conclusions The Cyclin D1 cDNA was obtained.Cyclin D1 was expressed in BL21.
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Objective To investigate the clinical application of fecal calprotectin in inflammatory bowel disease (IBD).Methods Colonoscopy took 79 patients with IBD that were diagnosed with pathology,including 47 cases of ulcerative colitis (UC) patients,32 cases of Crohn's disease (CD).Moreover,42 cases of IBD patients without abdominal pain,diarrhea and other intestinal inflammation were used as disease control group,and 34 cases of healthy people were used as healthy control group.The level of fecal calprotectin in each group was detected by enzyme-linked immunosorbent assay (ELISA).Results The positive rate of fecal Calprotectin in IBD group,disease control group and the healthy control group was 57.0％,19.0％,and 0,respectively; each positive rate in IBD group was significantly higher than the other two groups (P ＜ 0.05).The serum concentration of fecal calprotectin in IBD group [(493.86 ±204.18) μg/g] was significantly higher than the disease control group [(71.46 ± 60.51) μg/g] and the healthy control group [(36.19 ± 13.46) μg/g] (P ＜ 0.05) ; IBD active calprotection [(1015.23 ± 324.96) μg/g] was significantly higher than resting [(52.69 ±34.71) μg/g] (P ＜0.01).Conclusions Fecal calprotectin test benefits early diagnosis of IBD,and may be taken as the diagnostic index of IBD activity.It has extensively clinical value.
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Objective To investigate the value of urinary trehalase (T) in patients with renal proximal tubular damage.Methods 134 patients with kidney disease (male:66 female:68 age:18-59 ; 31cases with acute renal failure,30 cases with chronic renal failure,20 cases with drug-induced renal impairment,21 cases with renal transplantation and 32 cases with nephritic syndrome) and 101 healthy controls (58 males and 43 females) were selected.Urinary T,N-acetyl-D-glucosaminidase (NAG),β2-MG,gamma-glutamyl transferase (GGT) were detected.Data were analyzed by SPSS 11.5,including nonparametric rank test,ROC analysis.Results The level of urinary trehalase in control group was normally distributed (7.1 ± 4.1) μmol/h · g Cr (0-25 μmol/h · g Cr).There was no significant difference between men and women (t =0.63,P =0.53).Urinary T levels were significant higher in all kidney disease groups than in control group (Z =6.80,5.90,5.23,6.00,8.04,P ＜0.01).According to ROC curve,the area of urinary T under the ROC curve (AUC) in 134 patients was 0.9,significantly different with NAG,β2-MG,GGT area (P ＜ 0.01),the AUCs of T were 0.94,0.85,0.90,0.90,0.91 in acute and chronic renal failure group,drug-induced renal impairment group,renal transplantation group and nephritic syndrome group,respectively; Youden index were 0.85,0.65,0.77,0.66,0.72 respectively.Corresponding to the Youden index,sensitivity and specificity were 90.3％ and 95.1％,73.3％ and 92.1％,85.0％ and 92.1％,81.0％ and 85.2％,87.5％ and 84.2％ respectively.Conclusions The Urinary trehalase is better than other markers in the diagnosis of the proximal renal tubular damage.It was better to evaluate the proximal tubular function early in time.The diagnostic value of urinary trehalase played a key role in diagnosis,treatment and prognosis of kidney diseases.
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Objective To investigate the value of cystatin CysC on early renal damage in patients with essential hypertensive.Methods Hundred-four patients who were diagnosed as essential hypertensive with microalbuminuria (Urinary microalbumin:20-200 mg/L) with essential hypertensive (58 males and 46 females) were enrolled and 54 healthy subjects (30 males and 24 females) were selected as controls.Serum CysC (CysC)、Crea(Cr) 、BUN、uric acid (UA) were measured and ROC curve was established based on the examination.Results There were significant difference on the level of Serum CysC[1.22(0.91,1.51 ) mg/L vs 0.73 (0.61,0.79 ) mg/L,Z=3.30,P＜0.01],BUN [6.40 ( 4.43,9.06 ) mmol/L vs 5.10 ( 4.34,5.93 ) mmol/L,Z=5.94,P＜0.01],Cr [96.3 (72.6,122.0 ) μmol/L vs 70.5 (56.2,76.0 ) μmol/L,Z=8.30,P＜0.01],UA [375.7 ( 312.3,431.8 ) μmol/L vs 328.7 ( 271,379.3 ) mmol/L,Z=3.28,P＜0.01] between essential hypertensive group and control group.According to ROC curve,the area of CysC under the ROC curve (AUC) in 104 patients was 0.87,significantly different with CR(0.78),BUN(0.66),UA(0.66) (P＜0.05 or P＜0.01 ) The Youden index of CysC was 0.69,and the corresponding sensitivity and specificity of CysC were 76％ and 93％ respectively.Conclusion The diagnostic value of serum CysC on early renal damage in patients with essential hypertensive is superior to Cr,BUN and UA,and changes of renal function can be found earlier according to the level of serum CysC,It plays a key role in the diagnosis,treatment and prognosis of the early renal damage in patients with essential hypertension.
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To explore the association between polymorphisms of transcription factor 7-like 2 gene (TCF7L2) and type 2 diabetes mellitus in Hefei district. The results suggest that the genetic variation DG10S478 in the TCF7L2 gene was not associated with type 2 diabetes mellitus in Hefei district ( P>0.05 ). However, its attribution to the susceptibility of type 2 diabetes mellitus in Hefei is not important.
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Objective To study the treatment of sepsis caused by G - bacteria, anti-lipid A antibodies of bacterial endotoxin were screened from phage antibody library. Methods The mRNA was extracted from human B-lymphocytes against lipid A of bacterial endotoxin, reversely transcripted and amplified by polymerase chain reaction using general primers scanning Fd and light chain of IgG. The amplified fragments were inserted into pCOMB3 vector and electrotransfected competent E.coli XL 1-blue cells. Furthermore, the recombinant phage was lysed by coculture with helper VCSM13. Results Fab displayed on the surface as fusion protein with the N terminal of coat protein Ⅲ, and 4.8?10 6 clone library was established. Antibodies against lipid A of bacterial endotoxin were screened. Specific antibodies against lipid A of bacterial endotoxin were enriched by 100 times after three rounds of panning with lipid A.Conclusions Three clones exhibited specific binding to lipid A is identified by direct and competitive ELISA methods. The succcess of isolating anti-lipid A proves the usefulness of phage display system in human McAb preparation. The result shows that we have got the recombinant phage antibody.