Objective To construct recombinant adenovirns containing small interfering RNA (siRNA) targeting human VEGF-C mRNA,then to study the inhibitory effects of adenovirus-mediated VEGF-C siRNA on growth and lymphangiogenesis of human colon cancer in BABL/c nude mice model.Methods In vitro:the specific siRNA sequence targeting human VEGF-C mRNA was selected.The homologous double-strand DNA was designed and synthesized.After such DNA was inserted into pDC316-EGFP-U6 by BamHI and HindⅢ,pDC316-VEGF-C siRNA-EGFP-U6 was obtained,then it was co-transfected into 293 cells with the hone plasmid pBHGF35.After generated by homologous recombination,Ad5F35-VEGF-C siRNA-EGFP-U6 was obtained,and it was packaged and amplified in 293 cells.The human colon cancer model was established in nude mice by hypo-injection LoVo cells.They were divided randomly into four groups (n=6):adenovirus,virus without target gene,single siRNA and PBS control group.Injecting intervention intra-tumorly in each groups,calculating the tumors volume and drawing the growth curve,calculating micro-vascular density (MVD) and micro-lymphatic density (LVD) by staining respectively of the lymphatic and vascular with anti-LYVE-1 monoclonal antibody and anti-CD<,34> monoclonal antibody were completed.Results Adenovirus group tumor sizes were smaller than other groups.The LVD were 8.47±2.1 and 17.35±4.7 (P<0.05) in adenovirns group and the PBS control group.The MVD were 22.65±6.04 and 23.19±7.63 (P>0.05) in adenovims group and the PBS control group respectively.Conclusion The adenovirus-mediated VEGF-C siRNA can significantly inhibit the growth and lymphangiogenesis of human colon cancer in nude mice model,hut have no effect on microangium vessels growth.

Objective To observe the outcomes of one-stage posterior short-level pedicle screw fixation combined with anterior fixation of severe thoracolumbar fractures.Methods A retrospective case series study was performed on 21 patients with severe thoracolumbar fractures stabilized by posterior short-level pedicle fixation combined with anterior internal fixation at one stage from January 2012 to December 2014.There were 16 males and 5 females,at age of 17 and 64 years [(38.7 ± 11.4) years].The involved segments included T11 in 2 patients,T12 in 5,Lt in 6 and L2 in 8.For AO fracture classification,type A fractures were seen in 4 patients,type B in 7 and type C in 10.Thoracolumbar injury classification and severity score (TLICS) was (8.12 ± 0.87) points (range,7-10 points).Frankel neurological performance scale was Grade B in 8 patients,Grade C in 11 and Grade D in 2.Operation time,blood loss,nerve function,kyphosis correction and complications were reported.Results Operation time was (234.5 ±57.3)min (range,180-360 min),and blood loss was (387.4 ± 124.4) ml (range,260-950 ml).Time of follow-up was (19.8 ± 3.5)months (range,14-25 months).Nerve function of 18 patients was improved by at least one Frankel scale.Cobb angle was (4.1 ±5.3)° at postoperative 3 days and (4.0 ± 4.9)°at the final follow-up,showing significant differences from that before operation [(-9.3 ± 4.2) °] (P ＜ 0.05).While the difference of Cobb angle did not differ significantly at postoperative 3 days and at final follow-up.No cerebrospinal fluid leakage,vascular injury,incision infection or nerve function deterioration occurred.Conclusion One-stage posterior short-level pediele screw fixation combined with anterior decompression and bone graft fixation is characterized by short operation time,few blood loss,good correction of traumatic kyphosis and good neurological recovery,indicating a good surgical choice for severe thoracolumbar fractures.

To study the protective effect and the mechanism of asiatic acid (AA) from Potentilla chinensis on alcohol hepatic injury in rats. Male Wistar rats were randomly divided into six groups: the normal control group, the AA control group (8 mg · kg(-1) AA), the model group (5.0-9.0 g · kg(-1) alcohol) and high, medium and low-dose AA-treated groups (alcohol + 8, 4, 2 mg · kg(-1) AA). Each group was orally administered with the corresponding drugs once a day for 24 weeks. Approximately 1. 5 hours after the final administration, all rats were killed, and their blood samples and hepatic tissues were collected. The AST and ALT in rat serum and the contents of MPO, TNF-α, IL-1β, SOD, GSH-Px, GSH-Rd and MDA in hepatic tissues were detected. The expressions of NF-κB, TLR4, CD14, MyD88, TRIF and protein expression in hepatic tissues were measured by western blot. The pathological changes in liver tissues were observed by histological examination. The results showed that compared with the model group, the AA-treated groups showed significant decreases in serum ALT, AST and MDA and increases in the activities of SOD, GSH-Px, GSH-Rd and MPO. Moreover, AA markedly inhibited the expressions of TNF-α, IL-1β, TLR4, CD14, MyD88 and NF-κB. The histological examination showed alleviated hepatic issue ijury to varying degrees. In short, asiatic acid (AA) from P. chinensis could protect alcohol-induced hepatic injury in rats. Its mechanism may be related to the inhibition of NF-κB inactivation and the reduction of inflammatory response.
Animals ; Liver ; drug effects ; pathology ; Liver Diseases, Alcoholic ; prevention & control ; Male ; NF-kappa B ; physiology ; Pentacyclic Triterpenes ; pharmacology ; Potentilla ; chemistry ; Protective Agents ; pharmacology ; Rats ; Rats, Wistar ; Toll-Like Receptor 4 ; antagonists & inhibitors

OBJECTIVEAfter performed symptom-limited maximum cardiopulmonary exercise testing (CPET) before and after acute alkalized blood, we repeated CPET with pure oxygen.

METHODSFive volunteers, 3hr after alkalizing blood room air CPET, re-performed CPET inhaling from Douglas bag connected with pure oxygen tank. We compared with those of room air CPETs before and after alkalized blood.

RESULTSAfter alkalized blood oxygen CPET had a similar response pattern as those of CPETs before and after blood alkalization. During the CPET, all breath frequency, minute ventilation and tidal volume at each stage were similar to those of CPETs before and after alkalized blood (P > 0.05),except there was a lower peak tidal volume than those of both CPETs and a slightly higher resting minute ventilation only than CPET after alkalized blood (P > 0.05). After alkalized blood, oxygen CPET, all PaO2 and SaO2 and most Hb were lower than those of both CPETs (P < 0.05). The pHa and [HCO3-]a were higher than those of CPET before alkalized blood (P < 0.05); but were not CPET after alkalized blood (P > 0.05). PaCO2 was similar to that of CPET before alkalized blood (P > 0.05), but was lower than that of CPET after alkalized blood at resting and warm-up (P < 0.05); then was similar to both CPETs at anaerobic threshold (P > 0.05); but was higher at peak exercise higher than those of both CPETs (P < 0.01). Oxygen increased 2,3 volunteers' workload and time at AT and peak exercises.

CONCLUSIONRespiratory response pattern to oxygen CPET after alkalized blood is similar to those of both CPETs before and after alkalized blood. The CPET response is dominantly depended upon metabolic rate, but not levels of pHa, PaCO2 and PaO2.

Blood Gas Analysis ; Exercise Test ; Humans ; Oxygen ; Respiratory Physiological Phenomena

Objective: To evaluate the effect of penehyclidine hydrochloride (PHC) on Toll-like receptor 4 (TLR4)/nuclear factor kappa B (NF-κB) signaling pathway in non-ventilated lung injury in the patients undergoing radical operation for lung cancer. Methods: A total of 100 patients, aged 40-64 yr, with body mass index 18-27 kg/m², of American Society of Anesthesiology physical status Ⅱ or Ⅲ, undergoing radical operation for lung cancer, were divided into 2 groups (n=50 each) according to the random number table method: control group (group C) and PHC group.PHC 0.01 mg/kg was intravenously injected at 10 min before anesthesia induction in group PHC, while the equal volume of normal saline was given instead in group C. The peripheral tissues of the removed lung tissues were obtained for determination of wet/dry weight ratio (W/D ratio). The pathological changes and ultrastructure of lung tissues were observed under light microscope, and lung injury was assessed and scored.The expression of TLR4 and NF-κB protein and mRNA was detected by Western blot and real-time polymerase chain reaction, respectively.Before administration (T₀), at the onset of one-lung ventilation (T₁), at 60 min of one-lung ventilation (T₂), immediately after the end of one-lung ventilation (T₃), at the end of operation (T₄) and at 24 h after operation (T₅), blood samples were collected from the internal jugular vein for determination of serum tumor necrosis factor-alpha, interleukin-6 (IL-6) and IL-8 concentrations by enzyme-linked immunosorbent assay. Results: Compared with group C, the W/D ratio and lung injury scores were significantly decreased, the expression of TLR4 and NF-κВ protein and mRNA was down-regulated, and the concentrations of tumor necrosis factor-alpha, IL-6 and IL-8 were decreased at T₂-T₅ in group PHC (P<0.05). The pathological changes and damage to ultrastructure of lung tissues were significantly attenuated in group PHC as compared with group C. Conclusion The mechanism by which PHC attenuates non-ventilated lung injury is related to blocking TLR4/NF-κВ signaling pathway and reducing inflammatory responses in the patients undergoing radical operation for lung cancer.

The water of "J" lake has been seriously eutrophied; concentration of total nitrogen (TN), total phosphorus (TP) and chlorophyll a were all far above the 3rd level of the National Standard of Ground Water of China. The concentration of microcystin (MCYST) of the water at one site (M) was 1865 μg/l. There were 2.36μg MCYST-LR per mg dry waterbloom powder.

Objective To evaluate the efficacy and safety of recombinant adenovirus-p53(rAdp53) injection combined with radiotherapy and hyperthermia in the treatment of unresectable advanced soft tissue sarcoma.Methods In this retrospective study, we evaluated 76 patients with unresectable advanced primary or recurrent soft tissue sarcoma treated in our hospital from November 2005 to November 2012.These patients received radiotherapy and hyperthermia with rAdp53(p53 group, n=41) or without rAdp53(control group, n=35).rAdp53((1-2)×1012viral particles each time, once a week, 8 times on average) was injected into the tumor or infused into the pelvic cavity.Radiotherapy (2 Gy each time, 5 times a week) was performed for the planning target volume at 56.3±5.3 Gy in the p53 group and 58.1±4.2 Gy in the control group, with no significant difference between the two groups (P>0.05).Superficial or deep thermotherapy was employed 8 times on average (twice a week).Clinical features, response rate, time to progression (TTP), overall survival (OS), and adverse events were compared between the two groups (P>0.05).The Kaplan-Meier method was used to calculate OS;the log-rank test was used for survival difference analysis and univariate prognostic analysis;the chi-square test was used for comparison of categorical data.Results At 2 months after treatment, the p53 group had significantly increased response rate (partial response+ complete response+ stable disease)(85% vs.54%, P=0.003) and local control rate (49% vs.23%, P=0.020) as well as prolonged TTP (12 months vs.5 months, P=0.010) and OS (48 months vs.31 months, P=0.049), as compared with the control group.No adverse events caused by radiotherapy and hyperthermia except transient fever were seen in the two groups.Conclusions Concurrent radiotherapy and hyperthermia combined with rAdp53 injection is effective and safe for patients with advanced soft tissue sarcoma.

AIMTo study the pharmacokinetics and bioequivalence of acipimox sustained-release tablets (SRT) after a single and multiple oral dose in healthy dogs.

METHODSThe plasma concentrations of of SRT and reference capsules with a single and multiple oral doses.

RESULTSThe drug concentration-time profiles fitted to a noncompartment model. After a single dose administration of sustained-release tablets and capsules, the pharmacokinetic parameters were as follows: AUC were (158 +/- 30) and (147 +/- 37) microg x h x mL(-1); Tmax were (4.3 +/- 0.8) and (2.6 +/- 1.3) h; Cmax were (29 +/- 6) and (42 +/- 10) microg x mL(-1); T(1/2) were (2.3 +/- 0.7) and (1.60 +/- 0.10) h; MRT were (6.0 +/- 0.8) and (3.9 +/- 0.7) h, respectively. The relative bioavailability of the sustained-release tablet was (108 +/- 16) %. After a multiple oral administration of sustained-release tablets and capsules, the pharmacokinetic parameters were as follows: AUC were (209 +/- 23) and (195 +/- 26) microg x h x mL(-1); Tmax were (6.3 +/- 0.8) and (3.4 +/- 1.5) h; Cmax were (27 +/- 4) and (36 +/- 5) microg x mL(-1); Cmmin were (2.2 +/- 1.0) and (0.20 +/- 0.20) microg x mL(-1); Cav were (8.7 +/- 1.0) and (8.1 +/- 1.1) micro x mL(-1); FI were (293 +/- 73) % and (448 +/- 91) % , respectively. The relative bioavailability of the sustained-release tablet was (114 +/- 19) %.

CONCLUSIONThe results of two one-side test from single dose administration shown that two preparations were bioequivalent. The Cmax of sustained-release tablet was lower than that of capsules, while the Tmax and MRT of sustained-release tablet were higher than that of capsule, which indicating a good retarding effect. The results from multiple dose administration also shown that two preparations were bioequivalent and the DF of sustained-release tablet was significant lower than that of capsule.

Administration, Oral ; Animals ; Area Under Curve ; Biological Availability ; Capsules ; Delayed-Action Preparations ; Dogs ; Dose-Response Relationship, Drug ; Hypolipidemic Agents ; administration & dosage ; pharmacokinetics ; Pyrazines ; administration & dosage ; pharmacokinetics ; Random Allocation ; Tablets ; Therapeutic Equivalency

The three-step dissolution experiment was established to investigate the in vitro release of budesonide colon-specific tablet and to elucidate the drug release mechanism by fitting to different mathematical models. The physiological parameters of stomach, small intestine and colon such as pH value, intestinal flora, specific organic enzyme, vermiculation and conveying time were mimicked to plot the in vitro dissolution, separately. Sample were taken at predetermined time intervals in 24 h and the accumulated drug releases were determined by using HPLC method. Drug release curves of the localization tablets were fitted to various mathematical models. It shows that no drug release was found in 2 h. About 5% release was determined after 6 h while 77.5% accumulated release was reached within 24 h. Drug release from the in house formulation fitted well into first-order model. The three-step dissolution method could be used to evaluate the colon-specific characteristics of budesonide colonic localization tablet. The drug release behavior of the localization tablet conforms to the drug release mechanisms of controlled porosity osmotic pump where osmotic pressure is the main driving force for controlled delivery of drugs.
Animals ; Anti-Inflammatory Agents ; administration & dosage ; pharmacokinetics ; Budesonide ; administration & dosage ; pharmacokinetics ; Colon ; metabolism ; Delayed-Action Preparations ; Drug Carriers ; Drug Compounding ; Drug Delivery Systems ; methods ; Excipients ; Hydrogen-Ion Concentration ; In Vitro Techniques ; Intestinal Secretions ; Models, Theoretical ; Rats ; Tablets

OBJECTIVEBasis on the dynamic changes of the ventilation and arterial blood gas parameters to symptom-limited maximum cardiopulmonary exercise testing (CPET), we further investigate the effect of alkalized blood by drinking 5% NaHCO3 on ventilation during exercise.

METHODSAfter drinking 5% NaHCO3 75 ml (3.75 g) every 5 min, total dosage of 0.3 g/Kg, 5 volunteers repeated CPET. All CPET and ABG data changes were analyzed and calculated. At the same time, CPET and ABG parameters after alkalized blood were compared with those before alkalized blood (control) used paired t test.

RESULTSAfter alkalized blood, CPET response patterns of parameters of ventilation, gas exchange and arterial blood gas were very similar (P > 0.05). All minute ventilation, tidal volume, respiratory rate, oxygen uptake and carbon dioxide elimination were gradually increased from resting stage (P < 0.05-0.001), according to the increase of power loading. During CPET after alkalized blood, ABG parameters were compared with those of control: hemoglobin concentrations were lower, CaCO2 and pHa were increased at all stages (P < 0.05). The PaCO2 increased trend was clear, however only significantly at warm-up from 42 to 45 mmHg (P < 0.05). Compared with those of control, only the minute ventilation was decreased from 13 to 11 L/min at resting (P < 0.05).

CONCLUSIONEven with higher mean CaCO2, PaCO2 and pHa, lower Hba and [H+]a, the CPET response patterns of ventilatory parameters after alkalized blood were similar.

Blood Gas Analysis ; Carbon Dioxide ; Exercise Test ; Humans ; Oxygen ; Oxygen Consumption ; Respiration ; Respiratory Physiological Phenomena ; Tidal Volume