Objective To study the protective effect of leuprorelin particle on ovarian reserve function applied before the laparoscopic cystectomy of ovarian endometriosis.Methods 106 patients with endometriosis treated from January 2015 to December 2015 in our hospital were selected.The subjects were divided into two groups according to the random number table, 53 cases of each group.The control group adopted the laparoscopic surgery;the observation group took the leuprorelin particle before the laparoscopic surgery.The operation condition, ovarian hormone and function were observed. Results The operation time and bleeding amount in observation group was better than control group ( P <0.05 ) .After surgery, the exhaust time between two groups had no significantly different.After surgery, the level of follicle stimulating hormone (FSH) in observation group was lower than control group; the level of estradiol (E2) in observation group was higher than control group (P <0.05); the level of luteinizing hormone (LH) between two groups had no significantly different.After surgery, the ovarian volume and antral follicle count ( AFC) in observation group was better than control group (P<0.05).Conclusion Before the laparoscopic cystectomy of ovarian endometriosis, the leuprorelin particle protects the ovarian function and morphology and has less impact in the level of hormone.

The interaction betweenYan-Hu-Ning (YHN) and bovine serum albumin (BSA) was investigated in order to provide further theoretical evidences on action mechanism study between YHN and proteins within the organism. Under optimal conditions, the interaction between YHN and BSA was studied by fluorescence quenching, ultraviolet absorption spectrometry and synchronous fluorescence spectroscopy. At the temperature of 283.15 K, 298.15 K and 313.15 K, quenching constant (KSV) and speed constant (Kq) were calculated by S-V curves. Static quenching constant (KLB) was obtained by L-B double reciprocal equation. Double logarithmic equation was used to calculate the binding constants (Kb) and the number of binding site (n). Thermodynamic equation was used to obtainΔH,ΔS,ΔG. Hill’s coefficients (nH) was obtained by Hill equation. The results showed that at three different temperatures, along with the increasing of YHN concentration, the fluorescence intensity of BSA decreased regularly. The value of KSV, Kq, KLB, Kb, n and nH decreased with the increasing of temperature;ΔG < 0,ΔH < 0,ΔS < 0; n was approximately equal to 1; nH > 1. It was concluded that YHN-BSA complex quenched the intrinsic fluorescence of BSA. The mechanism of fluorescence quenching was static quenching. The main binding forces were deduced as hydrogen bonds and Van der Waals forces from calculated values of thermodynamic parameters. YHN and BSA can form a binding site, which indicated certain binding interaction between YHN and BSA. YHN can be stored and transported by protein within the body. Free energy was produced to transformΔG into negative value. It showed that the process of binding between YHN and BSA was spontaneous. The nH was more than 1. It indicated that YHN had positive cooperative effect. The primary binding site was located at subdomainⅡA. The synchronous fluorescence spectra showed certain influence on the conformation of BSA by YHN. It led to the weakening of polarity within BSA and the binding site to be closer to the tyrosine.

Background and purpose:Tumor microenvironment plays an important role in the introduction of foreign factors that mediate tumor acquired resistance. The antitumor effects of many chemotherapeutic agents depend on the level of oxygen pressure (pO2) in tumor microenvironment. This study aimed to evaluate electron paramagnetic reso-nance (EPR)-based monitoring on an oxygen-enriched tumor microenvironment to increase chemotherapeutic sensitivity. Methods:MCF-7 cells were used to establish human breast cancer in nude mice. EPR was used to directly measure pO2 levelin vivo. Tumor tissues were collected, and mitochondrial activity was assayed on the basis of the kinetics of enzyme-catalyzed reactions. A laser Doppler monitor was used to detect regional blood flow. Tumor apoptotic rate was analyzed by flow cytometry.Results:The tumor volume decreased more evidently in the chemotherapy group with oxy-gen-enriched environment than that in the conventional chemotherapy group after the treatment was administered (P<0.01). After chemotherapy was completed, the apoptotic rate of tumor cells was significantly higher in the chemotherapy group with oxygen-enriched environment than that in the conventional chemotherapy group (P<0.001). This study examined the mechanism ofpO2 changes in tumor microenvironment: This was related to the change of the balance between the oxygen consumption and the regional blood flow in the tumor tissues after chemotherapy.Conclusion:Based on the characteristics ofpO2 changes in the tumor microenvironment after chemotherapy was completed, the selection of chemotherapy mode forthe treatment inpO2 peak time window improves the sensitivity of chemotherapy, which provides a new idea for individual-ized chemotherapy in clinical applications.

Objective:To clone and express the Der f 7 recombinant protein from Dermatophagoides farinae and prepare the Der f 7 monoclonal antibody.Methods: The Der f 7 recombinant protein was expressed in prokaryotic expression system of pET28a(+)-Der f 7.BALB /c mice were immunized with the recombinant protein.Myeloma cells and spleen cells were fused,and hybridoma cells were screened by ELISA.Hybridoma cells were injected into the mice abdominal cavity to obtain ascites.It was purified by protein A agarose medium ascites,and then to dentified the titer and purity of the antibody.The specificity of the antibody was identified by Western blot.Results: Three hybridoma cells which stably secret recombinant Der f 7 monoclonal antibody were obtained.The monoclonal antibody had high purity,the titer was higher than 1∶243 000.Western blot showed that Der f 7 recombinant protein could be recognized well.Conclusion: We successfully obtained Der f 7 monoclonal antibody,which can be used for the quantification and localization of Der f 7 allergen and the diagnosis and treatment of allergic diseases.

Objective To investigate the change and clinical significance of serum hepcidin,serum ferritin (SF),transferrin re-ceptor (sTfR)and serum iron (SI)in patients in type 2 diabetes(T2DM).Methods 130 patients with T2DM were divided into 2 groups according to the 24 hour urine microalbumin (mAlb)quantitative:group A for trace microalbumin group 45 ca-ses (mAlb30~300 mg/24 h),group B for normal albuminuria group of 85 cases,an alternate period of 45 cases of healthy physical examination for group C (control group).Results Serum hepcidin and SF of group A (42.27±32.12 ng/ml,211.6 ±107.2 ng/ml)were significantly higher than those in group B (26.12 ± 18.36 ng/ml,179.1 ± 109.7 ng/ml)and the healthy control group (P <0.05),hepcidin and SF of group B was significantly higher than that of the control group (9.47 ±1.65 ng/ml,84.41±47.10 ng/ml,P <0.01),SI and transferrin receptor(sTfR)has no statistical significance between the three groups (P >0.05).Correlation analysis showed that patients with type 2 diabetes hepcidin was positively related with SF (P <0.05),hepcidin and sTfR,SI had no significant correlation.Conclusion These results indicated that there existed serum hepcidin and SF increased iron overload and iron metabolism disorder in type 2 diabetes.Therefore,detection of serum iron and SF can be used as a predictor of diabetes early renal damage.

Objective To compare the application of CUBIC and iDISCO clearing methods in observing 3D imaging of spinal cord with immunofluorescent staining. Methods 1 mm thick spinal cord coronal sections were processed with CUBIC and iDISCO, respectively. The neurofilament (NF) protein was detected by immunofluorescent staining and then was observed by a laser confocal microscope. Results Compared with CUBIC, iDISCO had the advantages of shorter time, higher transparency (F=6.64, P<0.01), and deeper penetration (F=5117.55, P<0.01). Conclusion Immunofluorescent staining combined with iDISCO could completely observe the spinal axons with shorter time and better stain effect.

Objective To observe the effects of neurotrophin 3 (NT3)-chitosan on motor function, and proliferation and differentiation of the neural stem cells (NSCs) in the injury area and subventricular zone (SVZ) in rats with motor cortex injury. Methods Sixty-five Wistar rats were divided into control group (n=7), injury group (n=29) and NT3-chitosan group (n=29). The motor cortex was aspirated and re-moved as cerebral injury model. NT3-chitosan was immediately implanted into the injured area after operation, and the control group re-ceived no intervention. Pellet reaching test was performed to detect the recovery of the forelimb function, HE staining was used to observe the lesion cavity size, and immunofluorescence staining was used to observe the proliferation and differentiation of NSCs 3 days, 7 days, 14 days, 1 month, 2 months and 3 months after operation. Results The grasp success rate was higher (F>6.00, P≤0.05), and the lesion cavity size was significantly smaller (F>629.5, P<0.001) in the NT3-chitosan group than in the injury group. In the NSCs differentiation experi-ment, the number of BrdU cells at all the time points was significantly higher in the NT3-chitosan group than in the injury group (F>171.43, P<0.001). In the NSCs proliferation experiment, the number of BrdU positive cells was still significantly higher in the NT3-chitosan group than in the control group and in the injury group (F>155.06, P<0.001), the number of Dcx positive cells was significantly higher in the NT3-chitosan group than in the injury group (F=62.367, P<0.001), and the number of BrdU/Dcx positive cells was significantly higher in the NT3-chitosan group than in the control group (F=33.527, P<0.001). Conclusion NT3-chitosan could activate NSCs in the SVZ, and pro-mote endogenous neurogenesis and forelimb function recovery in rats after motor cortex injury.

Objective: To investigate characteristics of intestinal flora in elderly patients with coronary heart disease (CHD) versus CHD plus complicated heart failure (HF). Methods: The 80 CHD patients admitted into our hospital from April 2017 to April 2018 were consecutively enrolled in this prospective study.They were divided into the CHD group (without HF, n=40) and the HF group (CHD plus HF, n=40), and 40 healthy elderly people taking health examination during the same period were considered as the control group.The 0.5 g stool specimen from each observer was collected.The specific DNA fragments of 16S rDNA/18S rDNA/ITS/ function genes were amplified, the Polymerase Chain reaction(PCR)amplified products were sequenced by Illumina Genome Analyzer IIx.And the sequencing results were followed by Read splicing, OTU clustering, alpha diversity analysis and species diversity analysis.Finally, the sample species information was obtained. Results: The mean abundances of gut microflora in CHD group and in HF group were lower than in control group (P<0.05). The abundance of phylum bacteroidetes was higher in the control group (66.41±3.25)% than in the CHD group (45.21±9.07)% and in HF groups (27.29±13.27)%(P<0.05). And the abundance of phylum firmicutes was lower in the control group than in the CHD and HF groups (P<0.05). LEfSe analysis showed that the flora with high abundances in the healthy elderly group were mainly Bacteroides, Bacteroidia and Bacteroidales.Ruminococcus and Clostridium occurred mainly in the CHD group, and Firmicutes and Enterobacte occurred mainly in CHD plus heart failure group. Conclusions The intestinal flora disturbance occurs in elderly patients with coronary heart disease or CHD with heart failure.

Transthoracic echocardiographic characteristics of 17 cases of cardiac amyloidosis (CA), a rare disease in China, were analyzed in order to improve the understanding of the disease. Seventeen cases of biopsy-proven CA, admitted to Wuhan Union Hospital from June 1994 to September 2008 were retrospectively reviewed. Twenty normal volunteers served as control group. Left atrial and ventricular functions and mitral inflow velocity were measured by two-dimensional, and Doppler echocardiography, and tissue Doppler imaging (TDI)-derived peak systolic wall motion velocities (Sv), peak early diastolic wall motion velocities (Ev), and peak late diastolic wall motion (Av) were measured at the septum, lateral, inferior and anterior corners of mitral annulus from the apical 4- and 2 chamber views. Compared with the control group, the interventricular septal thickness (IVSd), the left ventricular posterior wall (LVPWd), right ventricular transverse diameter (RVTDd) near the end of diastole and the interauricular septum thickness (IASs), left atrial anteroposterior diameter (LAADs), right atrial transverse diameter (RATDs) near the end of systole were increased significantly (all P<0.05) and left ventricular ejection fraction (LVEF) decreased (P<0.05) in the CA group. Compared with the control group, Sv, Ev at each wall and Av at almost all walls were significantly decreased in the CA group. In the CA group, Myocardial echoes of interventricular septum and free wall of left ventricle were enhanced evidently and distributed unevenly. The echoes presented as ground glass-like images, with some spotty hyper echoes. Both atria were enlarged, and LVEF decreased, with diastolic function impaired, and mild-moderate hydropericardium found in the CA group. It was concluded that echocardiography was a relatively sensitive and highly specific non-invasive method for the diagnosis of CA.
Amyloidosis/*ultrasonography ; Cardiomyopathies/*ultrasonography ; Case-Control Studies ; Echocardiography ; Echocardiography, Doppler/*methods ; Retrospective Studies ; Sensitivity and Specificity

Objective:To compare structure and function of mite Der f 3,Der p 3 and Eur m 3.Methods: Obtained mite allergens amino acid sequences from the International Union of Immunological Societies nomenclature database .Then physiochemical characterization,sequence alignment,secondary structure,three dimensional (3D) structure and epitopes of three proteins were analyzed by bioinformatics methods.Results:According to results of alignment ,Der f 3,Der p 3 and Eur m 3 displayed 88.51%sequence iden-tity.Der f 3,Der p 3 and Eur m 3 all contained three active sites and two trypsin functional domains ,which showed high identity of amino acid residues.Active sites of three proteins ,which closing to each other in three dimensional (3D) structure,constituting the active center of the enzyme.Secondary and 3D structure of three proteins all contains α-helices,β-sheets and random coils.Epitopes analysis revealed that Der f 3,Der p 3 and Eur m 3 all have 5 main potential epitopes located in random coils.Epitope sequences of Der f 3,Der p 3 and Eur m 3 overlapping in three domains (peptides of 79-81aa,129-135aa and 172-174aa),but the residues in these three domains were not identical.Conclusion:These studies lay the foundation for biochemical and genetic analysis of these 3 allergens,and may contribute to vaccine development for allergen-specific immunotherapy.