Objective To evaluate the influential factors of iron acquisition during Francisella tularensis LVS infection of mouse macrophages.Methods F.tularensis LVS expressing green fluorescent protein was used to infect murine macrophage J774A.1 cells.Transferrin receptor 1(Tfr1)was detected with mono-antibody and visualized with a goat-anti mouse IgG conjugated to Alexa 594.The expression profile of 5 iron metabolism related genes of J774A.1 murine macrophages uninfected or infected with F.tularensis LVS was determined with real-time PCR.Immunoblot analysis was used to compare the Tfr1 expression of live Francisella infected macrophage with dead bacteria.Tfr1 knock-off in J774A.1 cells was performed with siRNA.The transfected cells were infected with Francisella for immunoblotting and microscopy and infection assay.Results It was revealed that the live vaccine strain of F.tularensis induced the expression of Tfr1 in host macrophages.Gene expression analysis indicated that F.tularensis LVS drove an active iron acquisition program with induction of Tfr1 and iron regulatory proteins(Irp1 and Irp2).It was shown by Western-blotting that the siRNA-Tfrc-1 could knock off about 75% of Tfr1 in J774A.1 cells.It was determined by infection assay that,Tfr1 was knocked off,the bacteria number at 1h infection with Francisella was not different from that of control(F=1.06,P=0.326 5),while it was decreased significantly after 24h of infection(F=24.12,P=0.000 6).Conclusions It is demonstrated that upregulation of the Tfr1 may be mediated by post-transcriptional regulation during early infection,but sustained later through increased expression of Irp 1 and Irp 2.Increased expression of Tfr1 expands the intracellular iron pool through transferrin-mediated delivery and may thus be readily available for uptaking by Francisella.Knocking off the expression of Tfr1 does not affect bacterial invasion.Francisella,however,may fail to proliferate in macrophages in which the expression of transferrin receptor has been suppressed.

Based on the ongoing health reform and Wenzhou's economic and social developments,this article made a complete analysis on the policy packages initiated by Wenzhou government in August 2012,in an effort to encourage and involve private capital to launch medical institutions.These policies and measures released take into account the policy and legal environment for private capital in medical sector in China,and target the demands of deepening health reform and shortage of health development funding.Such efforts of Wenzhou are designed as breakthroughs in terms of market access,doctors mobility,return on investment,health insurance and fiscal policy.

In order to establish a method for the determination of the sterols of the oil in the freeze-dried acai (Euterpe oleracea Mart.) and to evaluate its antioxidant activities, a saponification/extraction procedure and high performance liquid chromatography (HPLC) analysis method were developed and validated for the analysis of phytosterols in PEE (Petroleum ether extract). Separation was achieved on a Purosper STAR LP C18 column with a binary, gradient solvent system of acetonitrile and isopropanol. Evaporative light scattering detection (ELSD) was used to quantify β-sitosterol and the total sterols. Peak identification was verified by retention times and spikes with external standards. Standard curves were constructed (r = 0.999 2) to allow for sample quantification. Recovery of the saponification and extraction was demonstrated via analysis of spiked samples. The highest content of total sterols is β-sitosterol. The antioxidant activities of the extracts were evaluated using the total oxyradical scavenging capacity assay (TOSC assay). The result showed that the PEE exhibited significant antioxidant properties, sample concentration and the antioxidant capacity had a certain relevance.
Antioxidants ; analysis ; Arecaceae ; chemistry ; Chromatography, High Pressure Liquid ; Drugs, Chinese Herbal ; analysis ; Phytosterols ; analysis ; Sitosterols ; analysis

BACKGROUND: Surgical guides designed based on a three-dimensional cone-beam CT (CBCT) model have been reported. However, CBCT cannot remodel fine soft tissue such as gums, and it can only be used to design a simple dental retainer with relatively poor stability. OBJECTIVE: To establish a high-precision three-dimensional (3D) integrated maxillodental model by matching CBCT model with 3D digital maxillodental model using 3D automatic registration method, based on which, we designed and manufactured individualized miniscrew surgical guides. METHODS: CBCT maxillodental models and laser-scanned dentition models obtained from six malocclusion cases were matched and overlapped using the 3D automatic registration method to fabricate the 3D integrated maxillodental model. Then, we accurately positioned and virtually implanted a micro-implant into the 3D integrated maxillodental model. Subsequently we prepared a high-precision individualized resin surgical guide by rapid prototyping technology. The inner diameter of the guide track was detected by a vernier caliper. Patients tried on the resin surgical guide, and then occlusion condition, guide seating and retention were detected. RESULTS AND CONCLUSION: Due to the high-precision registration of the model, all the resin surgical guide plates were suitable. The plate retention was enhanced after tooth clinching, and all the patients felt comfort when wearing the surgical guide plate, with no compression or other discomforts. The inner diameter of the guide track was (1.79±0.23) mm, and the measurement error was not statistically significant (P >0.05). These findings demonstrate that the high-precision surgical guide plate based on the high-precision 3D integrated model can provide the foundation for further investigations on the clinical application of surgical guides.

Objective To investigate the clinical factors influencing the treatment efficacy of microsurgery on patients with cerebral arteriovenous malformations (AVMs). Methods One hundred and eleven patients,admitted to our hospitals from January 1,2003 to December 31,2009 and underwent microsurgical cerebral AVMs resection were chosen in our study; their clinical data were retrospectively analyzed.The functional neurological status was classified with modified Rankin scale 7 d and 6 months after surgery. Binary logistic regression models were applied to test the effect of patient gender, age,intra-cerebral hemorrhage,and the size,deep venous drainage pattern and location of AVMs on long-term and short-term treatment efficacy. Results Twenty-nine patients (26.1％) showed new neurological deficits 7 d after the surgery.Univariate analysis and multivariate analysis demonstrated that deep venous drainage, eloquent location and large size of AVMs, high Spetzler-Martin grading aneurysms and no intra-cerebral hemorrhage were significantly associated with new early neurological deficits (P＜0.05),while gender and age of the patients had no significant association with new early neurological deficits (P＞0.05).The scores of neurological status scale in patients with Spetzler-Martin grading Ⅰ,Ⅱ and Ⅲ were significantly different between 7 d and 6 months after the surgery (P＜0.05). Conclusion AVMs patients having deep venous drainage,eloquent location and large size of AVMs,and having aneurysm and intra-cerebral hemorrhage may be the risk factors of new early neurological deficit after microsurgical resection; analyzing the Spetzler-Martin grading before surgery can help to determine the treatment options.

OBJECTIVETo establish a method suitable to determine the purgative biopotency of rhubarb and construct a new quality evaluation pattern of rhubarb.

METHODA series of factors such as observation index (mass of feces in 10 hours), animal strain (ICR mice), sex (male) and the dose of diphenoxylate complex (50 mg x kg(-1)) was investigated and fixed. The purgative biopotency as well as anthraquinone determination was used to evaluate the quality of different rhubarb samples.

RESULTThere wasn't a good linear relationship between the purgative biopotency and content of anthraquinone. The quality difference of rhubarb samples could be well characterized by combination of purgative biopotency determination and anthraquinone determination.

CONCLUSIONThe purgative biopotency determination can be used in quality control and evaluation of rhubarb.

Animals ; Anthraquinones ; analysis ; Biological Assay ; Cathartics ; pharmacology ; Dose-Response Relationship, Drug ; Drugs, Chinese Herbal ; chemistry ; pharmacology ; Female ; Linear Models ; Male ; Mice ; Quality Control ; Reproducibility of Results ; Rheum ; chemistry ; Sex Factors ; Time Factors

The essential issues of bioassay methods for quality control of traditional Chinese medicines (TCM) were investigated and discussed through the instantiation of developing the bioassay methods for laxative drugs. For the relatively broad variation of the quality of TCM, which might be influenced much by many factors, the parallel lines model of quantitative response is preferred to control the quality of TCM for its relatively high accuracy. The parallel model of quantal response is alternative while the sample can not meet the reliability standard for quantitative response model. According to the requirement of homogeneity between reference and sample for bioassay, the extract from referenced crude meterial is suitable to be used as reference substance after standardizing and defining by chemical substances, and to give reference to the establishment and reproducibility of authorized standard substance. The results of determination of the purgative biopotency of different species of rhubarb and the compound preparations showed that the bioassay methods and self-made reference substance established in this study could be used to control the quality of laxative medicines.
Animals ; Anthraquinones ; isolation & purification ; pharmacology ; Biological Assay ; methods ; Drugs, Chinese Herbal ; isolation & purification ; pharmacology ; Female ; Laxatives ; isolation & purification ; pharmacology ; Linear Models ; Male ; Medicine, Chinese Traditional ; Mice ; Quality Control ; Random Allocation ; Rheum ; chemistry

OBJECTIVETo evaluate the safety and efficacy of individual cylindrical abdominoperineal resection (CAPR) for locally advanced low rectal cancer.

METHODSFrom June 2011 to February 2012, 11 patients with locally advanced low rectal cancer underwent individual CAPR. There were 7 male and 4 female patients, aged from 32 to 74 years with a median of 64 years. Forty-seven patients underwent classic CAPR from January 2008 to February 2012. Preoperative and postoperative parameters such as clinical information of patients, tissue morphometry and complications were compared.

RESULTSIn the individual surgical group, 6 patients were treated with one side levator ani muscle totally or partially reserved, 3 patients with sacrococcyx reserved, and 2 patients with dissection close to the anterior rectal wall. Compared with classical surgery, the individual surgical specimens of horizontal section area ((2197 ± 501) mm(2)) and intrinsic muscle layer outer area ((1722 ± 414) mm(2)) were small, but the difference was not statistically significant (P = 0.150 and 0.167). The operative time, intraoperative blood loss, circumferential resection margin, total cross sectional tissue area, cross sectional tissue area outside the muscularis propria and bowel perforation rate between the two groups were not significantly different. Individual CAPR showed less incidence of chronic perineal pain (2/11, χ(2) = 6.116, P = 0.013) and sexual dysfunction (2/9, χ(2) = 4.412, P = 0.036) compared with classic CAPR.

CONCLUSIONSIndividual CAPR has the potential to reduce the risk of chronic perineal pain and sexual dysfunction without influencing the radical effect when compare with classic CAPR for the treatment of low rectal cancer.

Adult ; Aged ; Female ; Humans ; Male ; Middle Aged ; Perineum ; surgery ; Postoperative Complications ; epidemiology ; Rectal Neoplasms ; pathology ; surgery ; Rectum ; surgery ; Survival Rate ; Treatment Outcome

OBJECTIVETo explore the impact of abnormal myoelectricity at gastroduodenal anastomosis on gastric emptying in rats.

METHODSRats were randomly divided into experimental group (n=16) and control group (n=16). Pylorectomy and end-to-end gastroduodenal anastomosis were performed in the experimental group and electrodes were implanted in the serosal surface adjacent to the anastomosis. Slow waves were recorded by the implanted electrode in vivo. Gastric emptying was examined by scintigraphy.

RESULTSAt the first week after surgery, antral slow-wave frequency was significantly lower in the experimental group (0.8±1.4 vs. 3.3±1.2, P<0.01), as was the duodenal slow-wave frequency (2.1±0.6 vs. 11.1±0.7, P<0.01). There was no consecutive slow-waves transduction across the pylorus or the anastomosis. Within 12-16 weeks after operation, antral slow-wave frequency in the experimental group and the control group were (8.7±0.6) cpm and (4.0±0.4) cpm, respectively (P<0.01), and duodenal slow-wave frequency were (11.1±0.8) cpm and (10.8±0.7) cpm, respectively (P>0.05). Retrograde and antegrade myoelectricity transduction through the anastomosis were detected. The mean semi-emptying time in the proximal stomach was 14.7 min in the experimental group and 13.6 min in the control group (P>0.05). Radionuclide retention rate was 25.4% in the experimental group and 39.4% in the control group (P>0.05). The mean semi-emptying time in the distal stomach was 25.3 min in the experimental group and 10.5 min in the control group (P<0.01). Radionuclide retention rate was 46.4% in the experimental group and 18.7% in the control group (P<0.01).

CONCLUSIONThe abnormal myoelectricity in the region of gastroduodenal stoma may delay liquid gastric emptying in pylorectomy rats.

Animals ; Duodenum ; physiology ; surgery ; Gastric Emptying ; physiology ; Gastroenterostomy ; Male ; Myoelectric Complex, Migrating ; physiology ; Rats ; Rats, Sprague-Dawley ; Surgical Stomas ; physiology

Objective To investigate the value of introvoxel incoherent motion(IVIM)using 3.0 T MRI to evaluate response to concurrent chemoradiotherapy(CCRT)in patients with advanced uterine cervix cancer. Methods From July 2015 to December 2016,63 patients with advanced(≥ⅡB)cervical cancer diagnosed by clinical and imaging study, who had completed CCRT plan in Henan Cancer Hospital, were prospectively enrolled.Pelvic MRI protocol including T1WI,T2WI,IVIM and dynamic contrasted enhanced scans were performed in each patient before CCRT and 3 weeks after starting therapy(total dose of 30 Gy), and at the end of therapy (total dose of 90 Gy, 8 weeks after therapy). The mean values of ADC, true molecular diffusion coefficient(D),pseudodiffusion coefficient(D*)and perfusion fraction(f)in each tumor at pre-therapy, in the middle of therapy and post-therapy were measured and recorded as ADC-pre, D-pre, D*-pre,f-pre;ADC-mid,D-mid,D*-mid,f-mid and ADC-post,D-post,D*-post,f-post,respectively;the change rates of these parameters during and after therapy (recorded as ΔADC-mid, ΔD-mid, ΔD*-mid, Δf-mid;ΔADC-post, ΔD-post, ΔD*-post, Δf-post) were also calculated. Patients were classified into response group and non-response group,according to response evaluation criteria in solid tumors after CCRT.MRI imaging study was performed in each patient within 1 month after CCRT to follow up,and tumor regression rate was calculated.The Mann-Whitney U test was used to compare differences of parameters and their change rates between response group and non-response group. Spearman correlation analysis was performed to assess relationships between parameters, parameter change rates and tumor regression rate. Logistic regression model was applied to find potential ADC values for predicting therapeutic response. ROC was used to analyze efficacy of ADC values for evaluating therapeutic response in advanced uterine cervix cancer after CCRT. Results The mean value of tumor maximum diameter before and after therapy was (47.5 ± 12.9) and(12.8 ± 10.0)mm,tumor regression rate was(66.7 ± 33.6)%.Forty-eight patients were in the response group and 15 in the non-response group.The mean value of ADC-pre,D-pre,D*-pre and f-pre was 0.74(0.43, 1.14)×10-3,0.58(0.33,0.91)×10-3,12.12(2.30,21.4)×10-3mm2/s,9.65%(4.45%,13.89%),respectively.Tumor regression rate had positive correlation with ADC-pre and D-pre (r=0.773,0.840;P<0.05). Responders had increased ADC-pre, D-pre values than non-responders, which had statistically significant difference (P<0.05). Responders had increased ADC-mid, D-mid and f-mid values than non-responders, which had statistically significant difference (P<0.05), tumor regression rate had positive correlation with ADC-mid, D-mid and f-mid (r=0.808,0.834,0.563;P<0.05). Responders had increased ADC-post, D-post and f-post values than non-responders,which had statistically significant difference(P<0.05),tumor regression rate had positive correlation with ADC-post and D-post (r=0.799, 0.829;P<0.05).Tumor regression rate had positive correlation with ΔADC-mid,ΔD-mid,Δf-mid(r=0.526,0.573,0.454;P<0.05)and with ΔADC-post,ΔD-post, Δf-post (r=0.541, 0.555, 0.388;P<0.05). Responders had increased ΔADC-mid, ΔD-mid, Δf-mid and ΔADC-post, ΔD-post, Δf-post, which had statistically significant difference (P<0.05). Logistic regression analysis revealed only ADC-pre and D-post could be independent factors to predict therapeutic response in advanced uterine cervix cancer after CCRT,values of B,Wald,odds ratio and P was 22.488,8.431,1.429, 0.004 and 16.542,8.517,1.779,0.004.ROC analysis showed the area under the curve(AUC)of ADC-pre, D-pre,ΔADC-mid,ΔD-mid,Δf-mid,ΔADC-post,ΔD-post and Δf-post for predicting therapeutic response in advanced uterine cervix cancer after CCRT were 0.890,0.926,0.942,0.851,0.803,0.929,0.951 and 0.906, respectively. Conclusion The IVIM parameters before and during CCRT process and their changes are valuable for predicting and evaluating therapeutic response in advanced uterine cervix cancer after CCRT, with high clinical practice value.