Da Chaihu decoction is a classic prescription for the treatment of cholecystitis that is widely used in clinical practice, and has a definite curative effect. However, due to its diverse components and complex functions, the traditional indexes fail to capture its overall efficacy. Therefore, this study analyzed and predicted the quality markers (Q-markers) of Da Chaihu decoction based on specific chromatogram and network pharmacology to provide a reference for the comprehensive control of the quality. The study obtained 35 potential practical components of Da Chaihu decoction through virtual screening. The specific chromatogram of 15 batches of Da Chaihu decoction was established by HPLC-DAD with neohesperidin as a reference. Compared with the chromatographic peaks and the reference substance, the chemical components were assigned to predict the nine components of albiflorin, paeoniflorin, naringin, hesperidin, neohesperidin, baicalin, wogonoside, saikosaponin b₂, saikosaponin b₁ as Q-markers of Da Chaihu decoction. Finally, the network of the "components-key targets-signal pathways-biological processes" was constructed by network pharmacology to explore the mechanism of Da Chaihu decoction in treating cholecystitis to clarify the accuracy of Q-markers. The results indicated that potential Q-markers could act on multiple targets to regulate inflammatory and metabolism, and then combine to treat cholecystitis. Q-markers could combine with the pharmacologic action of Da Chaihu decoction, which could elucidate the overall efficacy of Da Chaihu decoction. This study explored the Q-markers of Da Chaihu decoction combined with the specific chromatogram and network pharmacology, which provided a basis for the quality control and evaluation of Da Chaihu decoction.

OBJECTIVE: To express and purify the antigenic peptide of adeno-associated virus (AAV) capsid conserved regions in prokaryotic cells and prepare its rabbit polyclonal antibody. METHODS: The DNA sequence encoding the conserved regions of AAV capsid protein was synthesized and cloned into the vector pET30a to obtain the plasmid pET30a-AAV-CR for prokaryotic expression and purification of the conserved peptides. Coomassie blue staining and Western blotting were used to identify the AAV conserved peptides. Japanese big ear white rabbits were immunized with AAV conserved region protein to prepare polyclonal antibody, with the rabbits injected with PBS as the control group. The antibody titer was determined with ELISA, and the performance of the antibody for recognizing capsid protein sequences of AAV1-AAV10 was assessed with Western blotting and immunofluorescence assay. RESULTS: The plasmid pET30a-AAV-CR was successfully constructed, and a recombinant protein with a relative molecular mass of 17000 was obtained. The purified protein induced the production of antibodies against the conserved regions of AAV capsid in rabbits, and the titer of the purified antibodies reached 1:320 000. The antibodies were capable of recognizing a wide range of capsid protein sequences of AAV1-AAV10. CONCLUSION We successfully obtained the polyclonal antibodies against AAV capsid conserved region protein from rabbits, which facilitate future studies of AAV vector development and the biological functions of AAV.
Animals ; Antibodies ; Capsid ; Capsid Proteins/genetics* ; Dependovirus/genetics* ; Prokaryotic Cells ; Rabbits ; Recombinant Proteins/genetics*

Objective To discuss the effects of polybrominated diphenyl ethers （PBDEs） exposure in e-waste dismantling region on the human body and provide data support for the identification of environmental health damage to residents in the e-waste dismantling region. Methods Adults in an e-waste dismantling region （exposed group, 54 participants） and a control region （control group, 58 participants） were selected, questionnaires were carried out and blood and urine samples were collected. Blood PBDEs, blood lipids, blood routine, blood lead, urine cadmium, urine chromium and urine nickel were detected. T-test was utilized to compare the differences of PBDEs between the two groups. Multivariate analysis were utilized to compare the differences between the two groups in blood routine indexes. Linear regression was used to analyze the relationship between PBDEs and blood routine. Results Exposure levels of PBDEs were significantly higher in the exposed group （240.00 ng/g, adjusted mass fraction of blood lipids, thereafter） than in the control group （93.00 ng/g, P<0.05）. There was no statistical significance in the differences in most blood routine indexes of the two groups （ P>0.05）, and their reference values were all within normal ranges. Mean platelet volume, plateletcrit, basophils percentage, absolute value of basophils, and mean corpuscular hemoglobin concentration were higher in the exposed group than in the control group （P<0.05）. Platelet distribution widths were lower in the exposed group than in the control group and below the normal reference range （P<0.05）. Conclusion PBDEs exposure in e-waste dismantling region tend to change platelet morphology, the number of basophils, and mean corpuscular hemoglobin concentration, and may pose potential health hazards to local residents.
Adult ; China ; Electronic Waste/analysis* ; Environmental Monitoring ; Halogenated Diphenyl Ethers/toxicity* ; Human Body ; Humans

Objective To evaluate the clinical value of maternal serum 25(OH)D level and bilateral uterine artery S/D mean in early prediction of pre-eclampsia(PE). Methods Sixty normal pregnancy women(normal group),40 mild preeclampsia women(MPE group)and 60 severe preeclampsia women(SPE group)who were examined in Changzhou First People′s Hospital and Changzhou Maternal and Child Health Care Hospital between January 2016 and June 2018 were included. The mean value of S/D of bilateral uterine artery was measured from 15th to 20th weeks in all the 3 groups,and serum 25(OH)D level of the mother was measured at 24th week. Meanwhile, the ROC curves of S/D mean value,serum 25(OH)D level and combined detection were drawn to compare the area under each curve(AUC),and the diagnostic efficiency of S/D mean value,serum 25(OH)D level and combined detection PE were also calculated. Results The incidence of adverse pregnancy outcome in SPE group was significantly higher than that in MPE group,and that in MPE group was significantly higher than that in normal group(P < 0.05). The mean value of S/D of bilateral uterine artery in SPE group was(4.09 ± 0.62),which was higher than that in MPE group(3.26 + 0.55)and in normal group(2.62 ± 0.51),while the mean value of S/D in MPE group was significantly higher than that in normal group and the difference was statistically significant(P < 0.05). The level of serum 25(OH)D in SPE group was(32.44 ± 5.84),which was significantly lower than that in MPE group(37.15 ± 5.90)and in normal group(42.57 ± 7.44),while the serum 25(OH)D level in MPE group was significantly lower than that in normal group,and the difference was statistically significant(P < 0.05). The mean value of S/D of bilateral uterine artery in the pre-eclampsia group was negatively correlated with 25(OH)D level(r = -0.66,P < 0.01). The area under the ROC curve separately detected by S/D mean value and 25(OH)D level was 0.787 and 0.719 respectively,both of which were lower than that by jointly detection(0.908)and the difference was statistically significant(P < 0.001). Conclusion Both the mean S/D value of bilateral uterine arteries and serum 25(OH)D level can be used for the diagnosis and monitoring of pre-eclampsia,and the diagnostic efficacy of the combined detection is superior to that of single detection.

Objective To understand the situation of college students' knowledge, attitude and practice (KAP) of innovation and entrepreneurship activities and analyze their influencing factors, in order to provide a scientific basis for further promotion of innovation and entrepreneurship activities. Methods A self-administered questionnaire survey using cluster sampling method was conducted among 850 undergraduates in a medical university in May, 2018. The demographic characteristics and questions pertaining to the KAP of innovation and entrepreneurship activities were collected, The weighted scores of KAP were calculated using the Delphi method. Chi-square test, multivariate logistic regression and linear regression analysis were used to analyze the data. Results A total of 850 questionnaires were sent out and 800 valid questionnaires were collected. The average scores and pass rates for the knowledge, attitude and practice of the 800 college students were (0.54 ±0.14), 38.1％; (0.56 ±0.23), 51.5％; and (0.37 ±0.19), 12.6％, respectively. Knowledge: the pass rate for the concepts related to innovation and entrepreneurship was 70.5％, and the pass rate for the policies related to innovation and entrepreneurship was 17.2％. The pass rate of senior students and excellent students were higher than those of junior students and students of lower academic competence (P=0.000, P=0.004). Attitudes: the pass rate for attitude towards the relationship between innovation and entrepreneurship and personal development was 63.6％, the pass rate for attitudes towards participation in innovation and entrepreneurship was 49.8％, and the pass rate for attitudes towards the school continuing to carry out innovation and entrepreneurship related activities was 23.8％. Practice:the pass rates for participation in scientific research and academic activities and social practice , voluntary cultivation of innovation ability and self-employment were 8.5％, 62.9％, 11.9％and 27.3％, respectively. The pass rate of senior grades was higher than that of junior grades (P=0.001). Scores for knowledge and those of attitude are positively correlated. Scores for practice and those of knowledge and attitude are positively correlated. Conclusion The higher pass rates for medical college students' attitude toward innovation and entrepreneurship activities as compared to those for knowledge and practice indicates high eagerness toward participation in innovation and entrepreneurship activities; a high rate knowledge is conducive to the cultivation of attitudes and the improvement of participation in activities . Therefore , it is necessary to strengthen the efforts in carrying out innovation and entrepreneurship activities , improve publicity and education, and promote the awareness-raising of innovation and entrepreneurship.

[Objective]To explore the evaluation value of ultrasomics based on contrast-enhanced ultrasound (CEUS)imaging in the therapy response of microRNA-122(miR-122)in hepatocellular carcinoma(HCC).[Method]Mice bearing subcutaneous HCC xenografts were injected intratumorally with microRNA-122 mimics(miR-122 mimics) and negative control mimics(NC mimics)in treatment group(n=6)and control group(n=6),respectively. The injec-tions were performed every 3 days for five times.Before each injection,two-dimension ultrasound(2D-US)imaging was performed.At 24 h after the last injection,2D-US and CEUS images of tumors were acquired,and then mice scarified for tumor miR-122 expression analysis by qRT-PCR.To evaluate the therapy response by RECIST,tumor volumes were mea-sured based on each 2D-US image. To analyze the tumor perfusion by mRECIST,perfusion parameters(maximum of intensity,rise time,time to peak,mean transit time,quality of fit)were analyzed off-line based on dynamic CEUS videos using SonoLiver?software. For ultrasomics,CEUS images at 10,30,60,90 second were used for features extraction, respectively. The corresponding ultrasomics formulas were built to evaluate the therapy response for miR-122.[Result]The tumors treated with miR-122 mimics resulted in a(763±60)folds increase in miR-122 levels compared to the tumors in control group(P<0.05).Effectively therapeutic response evaluated by tumor sizes change was detected after the third injection(P<0.05).For assessment using mRECIST,all the parameters of treatment group did not show significant difference from the ones of control group(P>0.05).Analysis using ultrasomics fail to detect different features of the static images of CEUS at 10 s,and models can be successfully built based on the rest of the three phases of CEUS images.The ultrasomics Scores between control group and treatment group were statistically different(P<0.05).The ultrasomics score at 30s were significantly lower than those at 60 s and 90 s,while there was no statistical difference between scores at 60 s and 90 s.[Conclusion]Ultrasomics analysis based on CEUS imaging is a useful method in evaluating the therapy response of miR-122 in HCC,and showed greater value than dynamic perfusion parameter.

Objective:To analyze whether the OAZI-1 (ornithine decarboxylase antizyme inhibitor-1) protein complex isolated from tumor cells could induce specific antitumor effects in the experiment mice .Methods:OAZI-1 protein complexes were isolated from B16-F1 melanoma cells by immune magnetic beads coated with OAZI-1 antibody and used as the vaccine to immune the C 57BL/6 mice.After immunization,the mice were inoculated subcutaneously with live B 16-F1 cells and then tumor formation and growth were ob-served.ELISA was used to determine the level of cytokine IFN-γin the serum of immunized mice.Lactate dehydrogenase assay (LDH) was performed to evaluate killing effect of spleen lymphocytes on B 16-F1 cells.The mice immunized by purified OAZI-1 from prokaryotic expression and PBS were used as controls in the animal experiment .Results: Compared with the control mice ,the spleen lymphocytes ( effector cells ) from the mice inoculated with OAZI-1 protein complexes had stronger killing ability on B 16-F1 cells (target cells).At three different effector:target ratio (10:1,50:1,100:1),the killing ability of these spleen lymphocytes were 46.2%, 59.5%and 92.5% respectively,which was significantly higher than the spleen lymphocytes from the mice inoculated with purified AZIN-1 protein (36.1%,26.8% and 45.9%) or inoculated with PBS (24.6%,24.0% and 27.2%).In addition,the content of serum anti-tumor cytokine IFN-γwas also significantly higher in the mice inoculated with OAZI-1 protein complexes (538.3 pg/ml) than the mice inoculated with purified AZIN-1 ( 256.2 pg/ml ) or with PBS ( 131.0 pg/ml ) .When B16-F1 live cells were subcutaneously inoculated into the immunized mice described above ,the tumor formation rate was only 40%in the mice immunized with OAZI-1 protein complex ,but 100%in the mice immunized with PBS or purified OAZI-1.The growth of inoculated tumors in the mice immunized with OAZI-1 protein complex was also much slower than the control mice .Conclusion:The results in this study suggest that the OAZI-1 protein complex isolated from B 16-F1 tumor cells could contain some tumor antigens .When used as tumor vaccine to inoculate mice ,this complex can induce anti-tumor immune killing activity in experimental animals .

Aim To investigate the influence of down-regulating adenosine A1 receptor and adenosine A2 A receptor gene expression on proliferation and activation of acetaldehyde-induced hepatic stellate cell-T6 cells through siRNA. Methods Alcoholic liver fibrosis in vitro model was constructed by inducing HSC-T6 cells with acetaldehyde. siRNA targeting A1R and A2AR were designed and synthesized according to its mRNA. The siRNA was transfected into rat HSC-T6 cells by li-posome LipofectamineTM 2000. HSC cell proliferation was measured by MTT. The mRNA levels of A1R, A2AR, α-SMA, Collagen I in the supernatant of the cell culture were measured by Quantitative Real-Time PCR. The protein levels of A1R, A2AR, α-SMA, Collagen I were measured by Western blot. Results A1 R and A2 AR siRNA effectively inhibited the cell proliferation, and they also significantly decreased the levels of A1R, A2AR,α-SMA, Collagen I, suggesting that A1 R and A2 AR might be potential target genes in the alcoholic liver fibrosis. Conclusions Silencing A1 R or A2 AR by RNAi can significantly inhibit the HSC proliferation, A1R and A2AR may be potential therapeutic target genes for alcoholic liver fibrosis.

AIM: To measure the macular thickness of normal young people by 3D 1000 optical coherence tomography (OCT) and study the repeatability of measuring results and the relationship between the thicknesses of macular and gender. At the same time, to compare our result with the data of other types of OCT, and to understand the consistency of the measuring results of macular thickness of different types of OCT.
METHODS: Totally 222 eyes in 111 young people were detected using 3D scan mode of Topcon 3D OCT 1000 (ver 2.4 ) . Twelve cases ( 24 eyes ) underwent repeatability check. We took transverse comparison between our measured results with other research's results.
RESULTS: There were 111 cases of young people, whose age were from 18-27 years old, all uncorrected and corrected visual acuity were≥1. 0, all intraocular pressure were <21mmHg. The average thickness of all macular region was 273. 32±17.08μm. Retinal volume of macular area was 7. 73 ± 0.37mm3 . Center thickness was 161 -264μm, and the average thickness was 200. 13±18. 81μm. Central macular thickness were 188 - 273μm, and the average thickness was 229. 00 ± 18. 20μm. The central macular thickness in men was significantly greater than that in women, and there was statistical difference. The results of repeated check of 12 cases ( 24 eyes ) in the macular area were no statistical difference except the outer ring of nasal quadrant, and the repeatability of average thickness in central macular thickness was better than in center thickness.
CONCLUSION:The repeatability of macular examination is good. The central macular thickness can be better repeated than the center thickness. The central macular thickness is 229. 00±18. 20μm in young people, according to the 3D 1000 OCT measurements. There are statistical difference of central macular thickness between different genders.

BACKGROUNDDiabetes has been associated with increased risk of fracture and impaired fracture healing. The aim of this study was to examine the influence of diabetes on perioperative complications, length of stay and ambulatory ability recovery in individuals with hip fracture, and to determine whether changes could be made to improve treatment outcome.

METHODSThe study included 707 hip fracture patients treated at Beijing Jishuitan Hospital between July 2009 and December 2010. The medical history and perioperative complications were compared between non-diabetic and diabetic groups. Length of stay, days awaiting surgery, and days of hospitalization after surgery were also analyzed. Ambulatory ability was compared at 1-year follow-up using the Chi-square test and Fisher's exact test. An independent Student's t-test was used to compare normally distributed continuous data.

RESULTSPatients with diabetes were more likely than non-diabetic patients to develop cardiac perioperative complications (8.9% vs. 3.0%, P = 0.021), urinary tract infections (12.0% vs. 2.8%, P < 0.001), and gastrointestinal symptoms (15.0% vs. 6.8%, P = 0.003). No difference in perioperative complications was observed between the groups. Days awaiting surgery and length of hospital stay were both longer in the diabetic group ((8.0 ± 5.1) vs. (6.2 ± 3.7) days and (16.5 ± 3.8) vs. (13.3 ± 3.8) days, P < 0.001, respectively). Before the occurrence of fracture, patients with diabetes were less likely to be ambulatory outdoors (71.9% vs. 85.9%, P < 0.001) and had more restricted walking ability. After at least 1-year follow-up, similar proportions of patients in the non-diabetic and diabetic groups (16.1% and 15.9%, respectively), who were able to ambulate outdoors before the fracture, became housebound till the final follow-up.

CONCLUSIONSDiabetics are at increased risk of specific complications and have a longer time to surgery and longer in-hospital stay, but generally have similar recovery to non-diabetics thereafter.

Aged ; Aged, 80 and over ; Chi-Square Distribution ; Diabetes Mellitus ; physiopathology ; Female ; Hip Fractures ; physiopathology ; Humans ; Male ; Middle Aged ; Perioperative Period ; Treatment Outcome