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We explored the sample collection and DNA extraction method of environmental DNA (eDNA) technique. Droplet digital PCR (ddPCR) was used to detect the eDNA concentration of the four major Chinese carps in different water layers and different cross-sections of Yichang section of Yangtze River trunk stream. We analyzed the relationship between eDNA concentration and density of egg and fry, and explored the feasibility of using eDNA technique to monitor the natural reproduction of the four major Chinese carps. Compared with the results from traditional survey methods, the eDNA concentration of the four major Chinese carps was extremely significantly correlated with egg and fry density. The maximum concentration was consistent with the peak occurrence time of egg and fry, suggesting that eDNA technique could predict the fishcrowd behavior for spawning. The investigation of cross-section eDNA concentration during two consecutive years suggested that the range of fish spawning ground was 4.5 km upstream to 1 km downstream of Yuyangxi. The predicted spawning ground was located within the range estimated by traditional methods from the lower part of Carmine Dam to Honghua Tuo. Our results indicate that eDNA technique, as a new ecological investigation method, could more accurately determine the distribution of fish with a certain population size, and thus with a good application prospect.

An in-depth understanding of the source of pollutants and the evolution of water quality is a prerequisite for water pollution control in source water reservoirs. Lijiahe Reservoir is an important water source for Xi'an. To strengthen the protection of Lijiahe Reservoir, water quality monitoring was carried out monthly from December 2016 to December 2017. The results showed that the main pollutant source was upstream water, and its percentage contribution to the bulk pollutants was 99.52% for permanganate index, 99.41% for TN, and 99.23% for TP. The main source of pollution in the upstream Dongcaiyu was the upper reach with Gepai Primary School and the turf service area. The main source of pollution in Xicaiyu was the upper reach with a chicken farm and fishery. In summer and autumn, Lijiahe Reservoir exhibited thermal stratification. During the stable stratification period, the maximum concentrations of TN, TP, permanganate index, TOC, Fe, and Mn in the water body were as high as 3.32, 0.177, 5.21, 3.01, 0.21, 0.235 mg·L -1 , respectively, under the dual effects of exogenous pollution and bottom anaerobic release. The maximum concentrations of TN, TP, Fe, and Mn exceeded the Class Ⅲ water limit of Surface Water Environmental Quality Standards (GB 3838-2002). The higher nutrient levels promoted large-scale reproduction of algae. The density of algae in the upper water reached 2.18×10 8 cells·L -1 , and the dominant algae species were Microcystis aeruginosa and Aphanizomenon flosaquae , which pose a significant threat to water quality. Therefore, the primary goal for water pollution control in Lijiahe Reservoir is to take effective measures to control algae blooms, and at the same time, reduce the input of upstream pollutants.

The present study was completed at the Lijiahe Reservoir in Xi'an, northwest China. Based on persistent monitoring of the physical and chemical indexes, in addition to phytoplankton activity, the correlation between phytoplankton distribution and dissolved oxygen depletion in metalimnion were analyzed during the thermal stratification periods. The results showed that:①During the summer stratification (from June to September), an obviously steady structure of dissolved oxygen, pH, as well as conductivity, was formed in a water column with RWCS/H indexes between 2 and 10 m -1 . ②The algal bloom was induced by the intense stratification with maximum biomass concentration of 2.95×10 8 cells·L -1 and chl.a of 46.42 mg·m -3 . With a sharp density gradient due to thermal stratification, the decayed algae cells accumulated above the metalimnion and largely consumed the dissolved oxygen, which affected the pH, conductance, as well as the permanganate index. ③Decomposition of high-density algae was, therefore, the main cause of metalimnion hypoxia during the summer stratification periods.

Aim: To clone and prokaryotically express the EGF and SP domain proteins of human mannan-binding lectin associated serine protease-2 (MASP-2), and prepare their polyclonal antibodies and analyze their immunogenicity., Methods: The cDNA of human MASP-2 was transcripted reversely from total RNA extracted from human fetal liver tissue, and then EGF and SP domain genes of MASP-2 were amplified from the cDNA by PCR. The EGF and SP fragment domain genes were cloned into pGEX-6P-2 expression vector and induced by IPTG to express GST fusion protein. The BALB/c mice were immunized with the purified fusion proteins to generate polyclonal antibodies. The specificity of the polyclonal antibodies was analysed with Western blotting, and the titer was determined by indirect ELISA., Results: We successfully constructed the recombinant pGEX-6P-2-EGF and pGEX-6P-2-SP and induced the expressions of the GST-EGF and GST-SP fusion proteins. The specificity of the polyclonal antibodies was higher, and had no cross reaction with other proteins. Indirect ELISA showed that the titer of anti-EGF antibody was more than 1:32 000 and anti-SP antibody was more than 1:40 000., Conclusion: The polyclonal antibodies of the EGF and SP domain proteins we successfully obtained were of higher specificity and titer, which provides the basis for further research on MASP-2.

Animal model of Pneumocystis carinii pneumonia (PCP) was established for acquiring lung tissue infected with P. carinii. After DNA from rat lungs was extracted, nuclear ribosome small subunit 18s rDNA of P. carinii was amplified by loop-mediated isothermal amplification method at 63 degrees C for 60 min. The product was digested by restriction enzyme Apal I. The results showed that 18s ribosome DNA (rDNA) of P. carinii was cloned into vector pGEX6p2, and the positive clones screened. Therefore, the loop-mediated isothermal amplification has been established for detecting P. carinii.

The expression and distribution of the imprinted paternally expressed gene 10 (PEG10) in decidual tissues of 36 patients with inevitable abortion and 36 normal pregnant (NP) women were examined by RT-PCR, immunohistochemistry, in situ hybridization and Western blot. Decidual PEG10 mRNA expression was significantly lower in the patients than that in the NP group by RT-PCR (P<0.05). The characteristic PEG10 expression was confirmed by in situ hybridization. As shown by Western blot and immunohistochemistry, PEG10 protein expression was consistent with the mRNA expression pattern. The results showed that maintaining sufficient levels of PEG10 may be important during early pregnancy. Lower levels of PEG10 might be one of reasons that underlie inevitable abortion.

Purpose: The purpose of this study is to develop a novel root canal filling sealers based on calcium phosphate cement (CPC), and to evaluate its physical-chemical properties and in vitro antibacterial activity on the predominant bacteria infecting root canal., Methods: The fluidity and the setting time of the sealer were tested according to ISO 6876:2001(E) standards. The crystal size of the final product was determined. Its opacification with different composition were measured. The in vitro antibacterial property of the sealer was tested according to the Antimicrobial Susceptibility Testing of Anaerobes recommended by National Committee for Clinical Laboratory Standards (NCCLs). The involved bacteria included Actinomyces naeslundii(A. naeslundii), Peptostreptococcus anaerobius (P. anaerobius), Porphyromonas gingivalis (P. gingivalis), Porphyromonas endodpntalis (P. endodpntalis) and Fusobacterium nucleatum (F. nucleatum). Twenty single-rooted human extracted teeth were selected to evaluate the sealing ability using dye microleakage technology. Dye penetration was measured and the results were statistically analyzed using SPSS12.0 software package., Results: The new root canal filling sealer was primarily composed of hydroxyapatite in 279 nm after setting. Its liquidity was suitable, the operating time was over 30 minutes, and the controlled setting time was (1.0+/-0.5) hours. The opacification was acceptable. MIC

Objective: To study the effect of Jingjielianqiao Decoction in promoting leg ulcer in rabbits., Methods: Nine adult male New Zealand albino rabbits with chronic leg ulcers were randomized into 3 groups, namely group A treated with Jingjielianqiao Decoction, group B with Shengjiyuhong Decoction, and group C with normal saline. Gross observation of the wounds was carried out regularly for evaluating the changes in the ulcerous area, depth and wound surface excretion. After 3 weeks of treatment, the tissues on the edge of the ulcer were sampled and prepared for routine pathological examination, electron microscopy and immunohistochemistry for vascular endothelial growth factor (VEGF) and CD34. The number of blood vessels and their areas were also recorded., Results: The wounds showed no significant differences between the 3 groups by gross observation during the treatment, but after completion of the 3-week treatment, routine pathological examination and electron microscopy revealed significant differences between the groups. Immunohistochemistry for VEGF and CD34 yielded comparable results between groups A and B (positive control), but showed significant differences between group C and the other two groups (P<0.01)., Conclusion: Jingjielianqiao Decoction and Shengjiyuhong Decoction can obviously promote the healing of leg ulcer in rabbits.

Objective: To survey the role of hepatocyte growth factor activator inhibitor type 1 (HAI-1) in early embryo development, we study the HA-1 gene expressing in villous tissue after embryo implantation., Methods: Fifty villous tissue samples from pregnant 6 w to 10 w were divided into five groups and each group included ten samples. The HAI-1 gene and protein expressing in the villous tissue were detected by RT-PCR and immunohistochemistry., Results: The expression of HAI-1 had a gradual increase with the pregnancy weeks added. The HAl-1 gene expression was very low at pregnant 6 w(A(260/280) 0.3142), From pregnant 7 w to 9 w, the expression was increased gradually (A(260/280) 0.3476 in 7 w and 0.3910 in 8 w) and reached the pinnacle in 9 w (A(260/280) 0.4876). And then the pinnacle value lasted until 10 w (A(260/280) 0.4909). The proportion of HAI-1 mRNA/beta-actin mRNA between 9 w and 10 w was not significantly different (P > or = 0.05). But among the other 3 groups, the proportion was significantly different (P < 0.01). The proportion of 9 w and 10 w was significantly different from the other 3 groups (P < 0.01). The results of HAI-1 protein expression showed the similar conclusion by immunohistochemical analysis., Conclusion: The HAI-1 gene may play an important role in early embryo development in human.

Objective: To study the pharmacodynamics of Jingjielianqiao decoction in promoting the healing of skin ulcer in mice., Methods: Mouse models of skin ulcer were established by inducing skin trauma and scald, respectively. The skin ulcer of the mice was treated with Jingjielianqiao decoction or recombinant human epithelial growth factor (rhEGF), and the ulcer area after treatment was compared between the two groups and with their respective self control. Inflammatory cell infiltration in the ulcer was observed under the microscope., Results: Compared with the self-control group, the skin ulcer area of mice treated with Jingjielianqiao decoction was reduced obviously (P<0.001), and the inflammatory cell infiltration also alleviated (P<0.001). Jingjielianqiao decoction produced significantly better therapeutic effect than rhEGF., Conclusion: Jingjielianqiao decoction can obviously improve the healing of skin ulcer in mice with better effect than rhEGF.

The nm23 gene was originally identified as a metastasis suppressor gene by differential hybridization of non-metastatic and metastatic clones of murine K-1735 melanoma cell lines. Subsequently, other highly homologous genes were characterized and shown to code for nucleoside diphosphate kinase (NDPK) isotypes. These enzymes have been evidenced in a wide variety of organisms. In human, up to eight distinct but highly related genes have been discovered. The nm23-H1 and nm23-H2 gene products share 88% identity and are about 95% and 98% identical to the murine nm23-M1 and nm23-M2 proteins respectively. A number of data indicate that nm23 family, initially documented as metastasis suppressors in some cancer types, are involved in the control of normal development, differentiation and apoptosis. The trophoblast cells of blastocyst and of the normal first-trimester placenta share some phenotypic similarities with malignant cells, such as rapid proliferation and ability to invade neighbouring tissues, including basement membrane, during the process of implantation and placental development, but do not have the ability for unlimited growth or metastasis, as few trophoblast cells can be identified in the deciduas, and trophoblasts rarely penetrate the maternal blood vessels. Although some data indicate that nm23 family play an important role in regulation of reproductive processes, little is know about its actual mechanism. In our studies, we performed RT-PCR, Western blot and immunohistochemical analyses of nm23-M1/NDPK A expression during the blastocyst adhesiveness in the mouse endometrium. The nm23-M1 mRNA expression was shown progressive increase in D5 mice endometrium by RT-PCR, which was consistent with the result obtained by Western blot and immunohistochemistry. It is suggested that nm23-M1/NDPK A was involved in the process of blastocyst implantation.

Leukemia inhibitory factor(LIF) is a glycoprotein with multiple activities, it expresses violently in most mice endometrium on 4-5 day of pregnancy,however the mechanism of LIF gene expression was not known completely. LIF gene expression was examined in endometrium of 168 mice and the concentrations of progesterone and estradiol were measured in their serum on 4-5 day. The expression of LIF gene was totally absent in 18 mice,and their concentrations of progesterone and estradiol in serum were significantly lower than the others. We discussed the relation between the expression of LIF gene with the concentration of two hormones. It was showed that the expression of LIF gene might be partly controlled by progesterone and estradiol.

To evaluate the relationship between habitual abortion of unknown etiology and expression of leukemia inhibitory factor (LIF) gene in the decidua,the expression of LIF gene in the decidua were detected in 32 cases of habitual abortion of unknown etiology (abortion group) and 36 cases of normal pregnancy (control group) by reverse transcription-polymerase chain reaction (RT-PCR). The results showed that the level of LIF mRNA expression of the decidua in abortion group was 0.512+/-0.219, significantly lower than those in the control group (1.314+/-0.457, P<0.01). The findings indicated that the low expression of LIF gene in abortion group might be one of the causes of habitual abortion. Analysis on the LIF gene expression of decidua can be used as a index for diagnosis of the habitual abortion.