Objective The key points in the epithelial-mesenchymal transition ( EMT) procedure include the downregula-tion of epithelial protein (E cadherin) and the upregulation of cell activity and cell matrix generation .The aim of this study was to es-tablish a method for primary culture and identification of mouse renal tubular epithelial cells and to explore whether the activation of C3aR can induce epithelial-to-mesenchymal transition in mouse primary renal epithelial cells . Methods Murine renal tubular seg-ments were used for primary cell culture .Immunocytochemistry and immunofluorescence staining were used to identify the renal tubular epithelial cells.The experiment groups included control group , five different concentrations of C3aR agonist groups (0.1, 1, 100, 500, and 2000 ng/mL), and three different time-point groups.The mRNA levels of E-cadherin,α-smooth muscle actin (SMA) and colla-gen I in renal tubular epithelial cells were detected by Real-time PCR; the protein of E-cadherin, α-SMA were detected by Western blot.The cytoskeleton of epithelial cells was observed by phalloidin staining . Results Compared with the control group , the protein expression of E-cadherin deceased (0.950±0.901 vs 0.650±0.221) and the expression of α-SMA (1.380±0.062 vs 1.600±0.103) and collagen I increased in C3aR agonist group (500 ng/mL, after 48 hours) (P<0.05).In addition, the association between these changes and C3aR agonists was presented in a dose-and time-dependent man-ner, respectively.The cytoskeleton staining showed that treatment of renal tubular epithelial cells with C 3aR agonists induced the formation of actin stress fibers in a time-dependent manner . Conclusion The method for primary culture and identification of mouse renal tubular epithelial cells were successfully established .The activation of C3aR could induce epithelial-to-mesenchymal transition in mouse primary renal epithelial cells , which plays an essential role in the de-velopment of renal fibrosis .Moreover , this study indicated that C 3aR may become a new therapeutic target in kidney diseases .

[Abstract ] Objective Evidence from previous studies indicated that over-activation of C3a/C3aR axis existed in the renal tubular epithelial cells of patients with renal diseases including diabetic nephropathy .However , the pathological significance of over-ac-tivating C3a/C3aR axis still remains to be elucidated .In this study, we constructed a renal tubular epithelial cell line over-expressing C3a in a secretory manner in order to provide a cell model to investigate the pathological significance of over -activating C3a/C3aR axis under various pathological scenes . Methods We designed a synthesized C3a secretory expression unit and cloned it into the multi-clonal site of lentivirus expression vector pLenti 6.3-MCS-IRES2-EGFP.After identification by sequencing , recombinant lentivirus was packaged by using pLenti 6.3-C3a-IRES2-EGFP and packaging plas-mid in 293T cells.Then, the recombinant lentivirus was used to in-fect HK2, a cell line of human renal tubular epithelial cells .After screening in medium with blasticidin , blasticidin resistant cell clones were obtained .Real-time PCR and ELISA method were applied to analyze the expression and secretion of stable transfected cells cloned C3a and identify renal tubular epithelial cell lines with stable over-activating C3a. Results ①C3a secretory expression unit was suc-cessfully synthesized and correctly cloned into the multi-clonal site of pLenti6.3-IRES2-EGFP; ②C3a secrectory expression recombi-nant lentivirus LV-C3a was successfully packaged with a high titer of 5 ×108/mL;③HK2 Cell clones resistant for blasticidin were ob-tained;according to the analysis of Real-time PCR and ELISA, the C3a mRNA level in HK2-C3a cell lines was significantly higher than that of HK2 cells(1.0 ±0.5 vs 1321.0 ±18.0, P<0.01) and the secreted C3a level increased significantly ([0.3 ±0.2]ng/mL vs [249.0 ±37.0] ng/mL, P<0.01). Conclusion The present study successfully constructed C 3a secretory expression vector pLenti6.3-C3a-IRES2-EGFP and C3a over-expression renal tubular epithelial cell line HK 2-C3a, which is very useful in further study of the function and significance of C 3a/C3aR axis not only in renal tubular epithelial cells but also in other cell types .

Objective Our previous study showed that the expression of C 3aR was increased in renal tubular epithelial cells (RTEC).However, the role of C3aR in diabetic nephropathy remains unknown, and so does the exact physiological and pathological signifi-cance of C3aR in relevant renal tissues.In the present study, we investigated the physiological and pathological significance of C3aR in RTECs using a lentivirus expression vector and anRTEC strain overexpressing C3aR. Methods Based on the sequence of C3aR mRNA, the human C3aR gene was synthesized and cloned into the multi-clonal site of the lentivirus expression vector pLenti6.3-MCS-IRES2-EGFP to construct a C3aR expression vector pLenti6.3-C3aR-IRES2-EGFP.After identification by sequencing, the recombinant lentivirus expressing C3aR was packaged by cotransfecting293T cellswith the recombinant lentivirusexpression vector and packaged plasmid .Then,the recombinant lentivirus was used to infect the human RTECline HK 2.After screening in the medium with blasticidin, blasticidinr-esistant cell clones were obtained, followed by identification ofthe human RTECline stably overexpressing C 3aR by real-time PCR and immunochemical staining. Results TheC3aR expression vector pLenti6.3-C3aR-IRES2-EGFP was successfully constructed and the sequence was proved to be correct. C3aR expression recombinant lentivirus was successfully packaged with a titer of 5×108/mL.Blasticidin-resistant HK2 cell clones were ob-tained and the expression of HK2-C3aR mRNA was significantly higherin the HK2-C3aR cells than in the non-transfected HK2 cells (2.33± 0.45 sv 1.00±0.09, P<0.05). Conclusion We successfully constructed a C3aR expression lentivirus vector pLenti6.3-C3aR-IRES 2-EGFP and a C3aR overexpression renal tubular epithelial cell strain HK2-C3aR, which is very contributive to further studies of the roles of C3aR in renal tubular epithelial cells and other types of cells.

OBJECTIVE To investigate the accumulated effect and the mechanism of repeated sc administration of acrylamide (AM) on learning and memory after 28 d,and the effect of AM on the amplitude of population spike(PS) potential after a single sc administration of AM. METHODS Female Wistar rats were sc adminstered with AM 10, 20 and 40 mg · kg-1 once a day for 28 d, and weighted every week. The ability of study and memory was evaluated, The morris water maze was used from 22nd to 28th day,followed by step down test on the 29th and 30th day. The escape latent period and the number of errors in those two days were recorded. Rats from normal control group and AM 40 mg·kg-1 group were taken to have their N-methyl-D-aspartate receptor (NR) and calmodulin-dependent protein kinaseⅡ (CaMKⅡ) protein levels detected by Western blotting. Additionally, some other female Wistar rats were sc administered with a single dose of AM 40 mg · kg-1 ,before the changes in PS potential amplitude induced by high frequency stimulation were recorded by long-term potential (LTP). RESULTS Compared with normal control group, the relative body mass gain was significantly decreased in AM exposure groups(P<0.01). Additionally, the escape latency period was significantly increased in AM 20 and 40 mg·kg-1 groups compared with normal control group, while the crossing frequency was not significantly different betmeen these four groups. Compared with the first day of step down test, the number of errors was significantly decreased(P<0.01) and the escape latency period was significantly extended(P<0.01) in normal control group on the 2nd day. However, the number of errors and the escape latency period did not significantly change in the AM groups between the two days. The results of Western blotting showed that the protein expressions and phosphorylation of NR2A and NR2B, as well as the phosphorylation of CaMKⅡ in AM 40 mg · kg-1 group were significantly increased compared with normal control group. LTP result showed that AM 40 mg·kg-1 significantly inhibited the amplitude of PS potential after a single percutaneous administration. CONCLUSION AM can inhibit the PS amplitude by inhibiting the release of glutamate, increasing the expressions and activities of NR,and inhibiting PS potential, thus affecting the hippocampal synaptic plasticity, and the function of learning and memory.

Objective To evaluate the use of preoperative MSCT in patients with clinically suspected colorectal disease after incomplete colonoscopy Methods After incomplete colonoscopy, 31 patients underwent MSCT before and after intravenous injection of iodinated contrast agent CT virtual colonography (CTVC), multiplanar reformation (MPR), shaded surface display (SSD), and Raysum images were obtained by using 4 different software in workstation The results of MSCT were compared with the findings of colonoscopy, intraoperative colon palpation, and postoperative pathology Results After incomplete colonoscopy, MSCT was successfully performed in all 31 cases Postoperative pathology revealed 22 colorectal cancers (one synchronous cancers, two with polyps) and 9 benign disease 22 colonic carcinomas were all correctly diagnosed by MSCT (including synchronous disease), 2 benign diseases were misdiagnosed as malignancy, and one polyp was missed Conclusion MSCT is a feasible and useful method for evaluating the entire colon before surgery in patients with occlusive disease

Objective:To investigate the effects of pre-pregnancy health examination combined with standardized health care during pregnancy on the pregnancy outcome at advanced maternal age.Methods:A total of 300 pregnant women who visited the Perinatal Clinic of the First Affiliated Hospital, School of Medicine, Shihezi University, China who received treatment between September 2018 and September 2019 were included in this study. They were divided into three groups according to pre-pregnancy health examination and heath care during pregnancy. In the observation group ( n = 78), pregnant women received pre-pregnancy health examination and health care during pregnancy on time and regularly. In the control group 1 ( n = 110): pregnant women did not receive pre-pregnancy health examination or regular health care during pregnancy. In the control group 2 ( n = 112), pregnant women received pre-pregnancy health examination or regular heath care during pregnancy. The pregnancy outcome was compared among three groups. The pre-pregnancy health examination and regular health care during pregnancy in women at advanced maternal age were correlated with gestational hypertension, gestational diabetes mellitus, premature delivery and other pregnancy risks. Results:There were significant differences in premature delivery (20.5%, 9.8%, 2.7%), gestational hypertension (23.1%, 12.5%, 3.6%), gestational diabetes mellitus (19.2%, 8.9%, 2.7%), premature rupture of membrane (17.9%, 10.7%, 3.6%), postpartum hemorrhage (12.8%, 8.0%, 1.8%), gestational anemia (19.2%, 8.0%, 2.7%) and placental abruption (7.7%, 8.0%, 0.9%) among control group 1, control group 2, and observation group ( χ2 = 16.047, 16.376, 14.724, 10.489, 8.781, 15.380, 6.744, all P < 0.05). The rate of cesarean section rate in the control group 1, control group 2, and observation group was statistically significant (70.5%, 44.6% and 27.3%, χ2 = 34.468, P < 0.05). There were significant differences in the incidence rates of macrosomia (15.4%, 8.9%, 2.7%), low birth weight infants (11.5%, 5.4%, 0.0%), stillbirth or late-term abortion (10.3%, 4.7%, 0.9%), neonatal asphyxia (16.7%, 7.1%, 1.8%) among the control group 1, control group 2 and observation group ( χ2 = 9.654, 12.840, 8.979, 14.284, all P < 0.05). Conclusion:The risk of adverse pregnancy outcomes is increased at advanced maternal age. Pre-pregnancy health examination combined with standardized health care during pregnancy can improve the pregnancy outcomes at advanced maternal age.

ObjectiveTo evaluate the significance of the detection of serum C-reactive protein (CRP) in elderly patients with diabetes mellitus and cerebral infarction.MethodsSeventy-two patients with diabetes mellitus and cerebral infarction (diabetic cerebral infarction group),66 patients with cerebral infarction and without diabetes mellitus (non-diabetic cerebral infarction group) and 60 healthy persons (control group) from October 2008 to January 2011 were selected.The level of serum CRP was detceted.ResultsThe level of CRP was ( 3.81 ± 2.23 ) mg/L in diabetic cerebral infarction group,( 2.48 ± 2.24 ) mg/L in non-diabetic cerebral infarction group and (0.68±0.16) mg/L in control group.The levels of CRP in diabetic cerebral infarction group and non-diabetic cerebral infarction group were significantly higher than that in control group (P ＜ 0.05).The level of CRP in diabetic cerebral infarction group was obviously higher than that in non-diabetic cerebral infarction group(P ＜0.05).CRP abnormal rate was 70.8％(51/72) in diabetic cerebral infarction group and 43.9％ (29/66) in non-diabetic cerebral infarction group,which had statistical significance (P ＜ 0.05).ConcluslonCRP has important predictive value to the occurrence and development of diabetic cerebral infarction.

Objective To identify the metabolic levels in prefrontal lobe in patients with post-concussion syndrome by proton magnetic resonance spectroscopy (1H-MRS), and to explore the relationship between metabolic levels and executive function. Methods The study was conducted in 40 patients with post-concussion syndrome and 20 normal controls. 1H-MRS on prefrontal lobe was performed in patients and controls, the NAA, Cho and Cr were measured and the ratios of NAA/Cr, Cho/Cr and NAA/(Cho + Cr) were determined. They were also evaluated executive functions by verbal fluency test (animal), Wisconsin Card Sorting Test (WSCT) and Tower of Hanoi (TOH). Results Compared with normal controls, the patients with post-concussion syndrome were significantly lower NAA/Cr and NAA/(Cho+Cr) ratios in left prefrontal lobe (P < 0.05). The NAA/Cr ratio in left prefrontal was significantly positive correlated with total scores of verbal fluency (r = 0.66, P < 0.05), categories of WSCT (r = 0.54,P < 0.05) and total score of TOH(r = 0.58, P < 0.05). The NAA/Cr ratio was significantly negative correlated with total errors (r = -0.53, P < 0.05) and persistent errors (r = -0.47, P < 0.05) of WSCT and mean executive time of TOH(r = -0.67, P < 0.05). Conclusions The metabolic levels of NAA in left prefrontal lobe in patients with post-concussion syndrome is significantly decreased , it is one cause of impaired executive functions.

Objective To compare the quality of prepared slices of Herba Ephedrae from different producing areas and to establish reference quality criterion of Herba Ephedrae.Methods Determined the content of Ephedrine and Pseudoephedrine in Herba Ephedra Prepared Herbal by HPLC.According to Chinese Pharmacopeia of 2000-year edition,extract,ash,water,impurity and foreign substance were detected; accelerated stability test was performed according to experience.Results The content of Ephedrine in Herba Ephedra is 0.995 %～1.589 %,honey-fried HerbaEphedra is 0.855 %～1.557 %;the content of Pseudoephedrine in Herba Ephedra is 0.560 %～2.087 %,honey-fried Herba Ephedra is 0.508 %～1.902 %; water-soluble extract was 8.83 %～18.30 %and 14.81%～27.45 %, alcohol-soluble extract was 7.74 %～18.83 %and 14.15 %～27.34 %, the total ash content was 6.49 %～10.29 %and 6.34 %～10.24 %and acid-insoluble ash was 0.19 %～0.42 %and 0.18 %～0.42 %in Herba Ephedrae and honey-prepared Herba Ephedrae respectively.The average water content was 8.10 %(s=0.3961)and 4.02 %(s=0.4674)and average content of impurity and foreign substance was 2.02 %(s=0.1954)and 2.01 %(s=0.2209)in Herba Ephedrae and honey-prepared Herba Ephedrae respectively.Conclusion This research will provide a reference criterion for the quality control of prepared slices of Herba Ephedrae.

Objective To assess the clinical significance of reverse trendelenburg position in preventing postoperative nausea and vomiting(PONV)in thyroid surgery.Methods 110 patients with papillary thyroid carcinoma(PTC) admitted from Feb.2013 to Mar.2014 were prospectively divided into experimental group and the control group according to whether reserve trendelenburg position was adopted.Univariate and multivariate method were used to analyze relations between PONV and surgical position,gender,age,body mass index,hypertension,surgical time,ASA classification,anesthesia,and motion sickness.Results Of the 110 cases of PTC,the incidence of PONV was 16.28％ (7/43) in the experimental group while it was 37.31％ (25/67) in the control group.The difference had statistical significance(P ＜ 0.05).Statistical analysis showed that PONV was related to patients'gender,surgical position,surgical time,and motion sickness,while only surgical position and motion sickness was the independent risk factors.Conclusion Reverse trendelenburg position surgery can help to prevent PONV and promote recovery.