AIM: To established an HPLC/MS/MS method for the study of pharmcokinetics of PMEA-Na (the mono-sodium salts of 9-[2-(phosphonomethoxy) ethyl] adenine) in beagle dogs. METHODS: PMEA-Na and internal standard 9-(3-phosphony-methoxypropyl) adenine were isolated from plasma by protein precipitation with methanol, and then analyzed adopting multiple reaction monitoring (MRM) mode. Using Xterra MS column, the mobile phases consisted of methanol:water:formic acid (25:75:0.5) at a flow rate of 0.25 ml·min-1. Beagle dogs received the intravenous dosage of PMEA-Na at 1.0, 3.0 and 6.0 mg·kg-1. Pharmacokinetic parameters were obtained from concentration-time curves by non-linear least-squares regression using the program DAS. RESULTS: The linear calibration curve was obtained in the concentration range of 0.02 to 20 mg·L-1 (r=0.999), and the limit of quantitition was 20 μg·L-1. The within-day and internal-day precisions (RSD) were less than 6.5% and 10.8%, respectively. The accuracy was 97.1%～107.3%. After a single dose studies in dogs the AUC were 2.3±0.5, 8.2±1.3 and 18.5±1.3 mg·L-1·h; the t1/2 were 3.9±1.8, 8.4±1.5 and 8.9±0.6 h; the CL were 0.44±0.09, 0.35±0.05 and 0.31±0.03 ml·h-1·kg-1 at the dose level of 1.0, 3.0 and 6.0 mg·kg-1 respectively. CONCLUSION: The analytical method is sensitive and specific for investigation the pharmacokintics of PMEA-Na in beagle dogs.

Aim To investigate the effects of(S,S)ZX-5 and(R,R)ZX-5 on eNOS mRNA and protein levels.Methods Cell culture was used for the experiments in vitro.The effects of eNOS mRNA were investigated by RT-PCR.The effects of eNOS protein levels were investigated by Western blot.Results(S,S)ZX-5 in concentration of 30 mg?L-1 up-regulated eNOS expressions at protein and mRNA levels significantly in cultured ECV304(P

Objective To discuss the treatment efficacy of the femoral-head osteonecrosis by umbrella strut bone grafting and bone marrow mesenchymal stem cells. Methods The femoral-head osteonecrosis models on one side were established in forty-two goats using liquid nitrogen, which were randomly classified into three groups after 8 weeks. Group A were conducted umbrella strut bone grafting and bone marrow mesenchymal stem cells transplantation, group B were conducted simple unit side muscle bone flap grafting, and group C were conducted umbrella strut bone grafting and Cancellous bone grafting transplantation. The health side of three groups was treated as self-control. The radiological and histology examination were made at 3,6treated lateral femoral-head shape restored,cystic cnlow-density area disappeared, the original collapse had repaired, and no further collapse of the femoral-head shaped for both group A and B. Bone fusion was good and bone column shadow had been vague for group A, while the bone fusion healed somewhat less and bone column shadow still clear for group B. Group C had irregular contour of femoral head, flat collapse, uneven density, and fracture cracks could be seen at weight-bearing area of femoral head, obviously was the poorest bone had different states: mature and arranged ruled for group A, childish and irregular arrangement for group B, thinning, sparse and fracture for group C. The percentage of empty lacunae counts decreased, while the percentage of trabecular bone area fraction increased for group A. In contrast with group B, and C, the difference was statistically significant (P ＜ 0.05), but the difference was no longer statistically significant when contrast with healthy sides (P ＞ 0.05). 6 months later, the radiological and histology examination showed that:group A and group B had been no significant difference comparing to normal femoral heads, while the femoral head of group C collapsed and disappeared completely. Group A and group B were no difference comparing the healthy side (P ＞ 0.05)in the area fraction of trabecualr bone and percentage of empty lacunae counts, neither it was between group A and group B(P ＞ 0. 05). Conclusion The treatment efficacy of the femoral-head osteonecrosis by umbrella strut bone grafting and bone marrow mesenchymal stem cells was good and was better than cancellous bone grafting transplantation and simple unit side muscle bone flap grafting.

Azone ( 1桪odeylazacyclohenptan-2-one) is one of the best skin penetration enhancers. It can increase the permeability of skin for some hydrophobic compounds, so that it is possible to be admi-nistrated through skin. The effects of Azone on permeability of skin of a-Asarone, Nicorandil and Nitroglycerin were studied by flow through diffusion cell with radioactivity tracer.The results showed that the different concentrations of Azone gave different enhancement. The penetration of the 3 drugs was enhanced about 1-fold by

AIM To provide toxicokinetics data for toxicity studies of repeated doses of sodium 9-[2-(phosphonomethoxy) ethyl] adenine (PMEA-Na). METHODS The concentrations of PMEA-Na in plasma and urine were determined by HPLC/MS/MS method after single and multiple iv administrations in dogs. Data were executed by the statistical moment method to acquire the toxicokinetics parameters. Serum biochemical tests and histopathological examination were performed. RESULTS The system exposure of PMEA-Na in dogs was dose-dependent over the dose range of 1.0-6.0 mg·kg-1. The areas under the plasma concentration-time curve of PMEA-Na after single and multiple iv administrations at 1.0, 3.0 and 6.0 mg·kg-1 dosage were (2.3±0.5), (8.4±1.6), (17.5±3.7) and (5.0±0.4), (15.9±3.2), (30.3±4.7)mg·L-1·h, respectively. The urinary excretion of PMEA-Na in 72 h after iv administration was (87.0±4.8)% at the dose of 3.0 mg·kg-1. In 6.0 mg·kg-1 dose group, liver enzyme activity of glutamic-pyruvic transaminase and serum levels of total bilirubin, blood urea nitrogen, creatinine and triglycerides were all significantly elevated; glucose level significantly decreased comparing with the control group. Histopathological observation showed distinct pathological changes in liver and kidney tissues of 6.0 mg·kg-1 dose group. CONCLUSION There was evidence of toxicity after repeated-dose (14 d) of PMEA-Na in dogs and the major toxicity target organs were the kidney and liver.

Objective: To investigate the relationship between the melatonin and the development of testicle in rabbits. Methods: Thirty male rabbits (unreach puberty) were randomly divided into 2 groups:15 were actively immunized with melatonin and complete antigen, which was successfully synthesized through Mannich reaction by combining melatonin with BSA, the other 15 were taken as controls.The specific antibodies of rabbits were detected by ELISA and RIA.The serum testosterone(T) and luteinizing hormone(LH) were detected by RIA, the mass of testicle or epididymis and the number of sperm were measured. Results: The serum T and LH concentration in experiment group were significantly lower than that in the control group.The mass of testicle or epididymis and the number of sperm in experiment group were also significantly lower than that in the control group. Conclusion: The results suggest that the immunization of melatonin can block the development of testicle in male rabbits.

Underactive bladder (UAB) belongs to the category of "urinary retention" and "urinary incontinence" in Traditional Chinese Medicine (TCM). Its pathogenesis is based on spleen qi deficiency and kidney deficiency, while dampness heat and blood stasis are the symptoms, and the nature of the disease is basedon the deficiency in nature and excess in superficiality and simultaneous occurrence of deficiency and excess syndrome. TCM treatment of UAB is mainly based on spleen qi deficiency syndrome, kidney deficiency syndrome, water and heat syndrome, deficiency and blood stasis syndrome to be treated with oral taken of Chinese materia medica, or the use of acupuncture, moxibustion, acupoint application and other external treatments, or the combination of acupuncture and medicine, TCM combined with rehabilitation training and other comprehensive treatments. All methods can improve the urodynamic indexes of patients, relieve symptoms and improve the quality of life, with certain efficacy.

OBJECTIVETo establish a method for detecting plasma concentration of corn polysaccharide iron complex (CPIC) and investigate its absorption, distribution and elimination in rats.

METHODSUsing radioactivity isotope tracing method, we detected the radioactivity of (59)Fe-CPIC in the plasma of rats at different time points by gavages of 3 doses (28.0, 14.0, and 7.0 mg/kg) (59)Fe-CPIC in SD rats. The pharmacokinetic parameters was obtained using DAS 2.0 program for analysis of tissue distribution and elimination of (59)Fe-CPIC.

RESULTSThe standard curve was linear within the range of 0.14-141 µg/ml (r=0.9999, n=5). The average recovery was 95% with a relative standard deviation no more than 15%. The pharmacokinetic parameters at 3 doses obtained, namely t1/2 and AUC (0-), were 214∓104, 231∓110, and 181∓81 min, and 1986.3∓513.3, 737.0∓467.0, and 315.1∓226.1 mg·min-1·L(-)1, respectively. (59)Fe-CPIC were detected in all the 13 tissues types examined and high radioactivity intensity was found in the gastrointestinal tract, hematogenic organs and other organs rich in blood. (59)Fe-CPIC was eliminated after intragastric administration primarily via the feces in rats.

CONCLUSIONThe method we established is easy and specific, and the pharmacokinetic parameters of (59)Fe-CPIC fit the two- compartment open model.

Administration, Oral ; Animals ; Area Under Curve ; Coordination Complexes ; administration & dosage ; pharmacokinetics ; urine ; Feces ; chemistry ; Female ; Intestinal Absorption ; Iron ; administration & dosage ; pharmacokinetics ; urine ; Iron Radioisotopes ; Male ; Polysaccharides ; administration & dosage ; isolation & purification ; pharmacokinetics ; urine ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Tissue Distribution ; Zea mays ; chemistry

Objective:To analyze the law of TCM syndrome differentiation and treatment for type 2 diabetic kidney disease (T2DKD) stage Ⅳ based on literature research.Methods:Literature on type 2 diabetic kidney disease stage Ⅳ was retrieved from CNKI, WanFang data, VIP and SinoMed database. The retrieval time was from the establishment of the databases to December 31, 2020. Data screening was conducted based on the inclusion and exclusion criteria prior to data entry in Microsoft Office Excel 365. Data mining and statistical analysis were performed by SPSS Statistics 23.0 and SPSS Modeler 18.1.Results:A total of 110 articles with 3 969 T2DKD stage Ⅳ cases, 111 prescriptions and 206 kinds of Chinese materia medica were included. Kidney and spleen were the main location of T2DKD stage Ⅳ. T2DKD stage Ⅳ based on TCM deficiency in nature syndrome was mainly based on qi and yin deficiency, and the most common excess in superficiality syndrome was blood stasis. The prescriptions commonly used included Liuwei Dihuang Decoction, Zhenwu Decoction, Buyang Huanwu Decoction, and Shenqi Dihuang Decoction etc. The classification of medication efficacy with the highest frequency was qi-tonifying herb, followed by blood-activating and stasis-resolving herb. Among them, Astragali Radix was the core Chinese materia medica in the prescription. The results of association rule obtained 54 association rules. Conclusions:The disease characteristics of T2DKD stage Ⅳ is simultaneous occurrence of deficiency and excess syndromes. The deficiency in nature is mainly characterized by deficiency of qi and yin, deficiency of spleen and kidney, deficiency of spleen-kidney yang, and excess in superficiality is mainly characterized by blood stasis, dampness and toxin. Tonifying qi and nourishing yin, activating blood circulation and dredging collaterals are the basic treatment methods, while strengthening spleen and kidney, dampness and detoxification should be emphasized. Astragali Radix, Angelicae Sinensis Radix, Salviae Miltiorrhizae Radix et Rhizoma, Poria, Dioscoreae Rhizoma, Corni Fructus, Rhei Radix et Rhizoma and Alismatis Rhizoma were the basic Chinese materia medica in this period, which reflects the idea of "treating qi, blood and water together".

Objective To investigate the effects of dexmedetomidine postconditioning on brain injury after cardiac arrest and resuscitation in a swine model.Methods Twenty-eight healthy male domestic pigs,weighing 36±2 kg,were randomized (random number) into 4 groups (n=7 each group):sham operation group (S group),cardiopulmonary resuscitation group (CPR group),low-dose dexmedetomidine postconditioning group (LDP group),and high-dose dexmedetomidine postconditioning group (HDP group).Animals in the S group only underwent the surgical preparation.In the other three groups,the experimental model was established by 8 mins of electrically induced ventricular fibrillation and then 5 mins of cardiopulmonary resuscitation.At 5 min after resuscitation,a loading dose of dexmedetomidine of 0.25 μg/kg was intravenously infused followed by continuous infusion at a rate of 0.25 μg/(kg·h) for 6 h in the LDP group,and a loading dose of dexmedetomidine of 0.5 μ.g/kg was infused followed by continuous infusion at a rate of 0.5 μg/(kg·h) for 6 h in the HDP group.The same amount of normal saline was administered in the S and CPR groups.At 1 h,3 h,6 h and 24 h after resuscitation,the levels of serum neuron specific enolase (NSE) and S100B protein were measured.At 24 h after resuscitation,neurologic deficit score (NSD) was evaluated.After that,the animals were euthanized and cerebral cortex was obtained for the determination of tumor necrosis factor-α (TNF-α),interleukin-6 (IL-6)and malondialdehyde (MDA) contents,superoxide dismutase (SOD) activity,cell apoptosis and caspase-3 expression.Results Compared with the S group,post-resuscitation neurologic dysfunction and brain injury were observed in the other three groups,which were indicated by significantly higher NDS and markedly greater levels of serum NSE and S 100B (all P＜0.05).Compared with the CPR group,the score of NDS at 24 h post-resuscitation were significantly lower and the levels of serum NSE and S100B at 6 h and 24 h post-resuscitation were significantly less in the LDP and HDP groups [NDS:194±26,103±16 vs 278±23 at 24 h;NSE (ng/mL):32.4±1.8,28.6±3.7 vs 36.2±2.8 at 6 h,39.9±4.2,35.1±1.5 vs 45.1±3.0 at 24 h;S100B (pg/mL):2 534±207,2 382±170 vs 2 825±113 at 6 h,3 719±164,3 246±176 vs 4 085±161 at 24 h,all P＜0.05].Compared with the LDP group,neurologic dysfunction and brain injury at 24 h postresuscitation were further significantly alleviated in the HDP group (all P＜0.05).Pathological analysis indicated that brain inflammation,oxidative stress and cell apoptosis were observed after resuscitation in the CPR,LDP and HDP groups.However,the contents of TNF-α,IL-6 and MDA were significantly lower while the activity of SOD was significantly higher,and cell apoptosis and caspase-3 expression were significantly reduced in the brain after resuscitation in the LDP and HDP groups compared with the CPR group (all P＜0.05).In addition,those pathological injuries mentioned above were further significantly alleviated in the brain after resuscitation in the HDP group compared to the LDP group (all P＜0.05).Conclusions Dexmedetomidine postconditioning significantly alleviated the severity of postresuscitation brain injury in a dose-dependent manner,in which the protection was produced possibly through reducing tissue inflammation,oxidative stress and cell apoptosis.