Objective To established a method for the determination of puerarin. Method The sample solutions were pretreated by solid phase extraction (SPE): the specimens were eluted and the cartridges saturated gradually by them in the SPE, but the strongly absorbed impurities were held by the cartridges all the same, and then the effluents were measured by HPLC. Results The linear range of puerarin was 0.205～ 2.464? g, r=0.9999. The average recovery of puerarin in Xintong Oral Liquid, Radix Purerariae and Ganmao Zhike Capsule were 99.5 % , 99.9 % and 98.5 % , RSD were 1.67 % , 1.45 % and 0.95 % (n=5) respectively. Conclusion The method is simple and accurate and can be used for the determination of puerarin in the Chinese patent medicines containing Radix Purerariae. 

AIM: To investigate the effects of myocardial remodeling of aged left atrium (LA) on atrial ar-rhythmogenesis in rabbits .METHODS:The male New Zealand rabbits were divided into young LA and aged LA groups . By observing the changes of monophasic action potential ( MAP) and burst-pacing in LA of the rabbits in vivo, the main cardioelectrophysiological parameters such as resting membrane potential ( RMP) , action potential amplitude ( APA) , ma-ximum rise veloctiy of action potential (dv/dtmax), plateau potential and action potential duration at 30%, 50%and 90%( APD30 , APD50 and APD90 ) , as well as the inducibility and duration of atrial arrhythmias were recorded .L-type calcium current (ICa,L) was analyzed via whole-cell patch-clamp technique in enzymatically dissociated single rabbit LA myocytes . The myocardial collagen content was quantified after Masson staining , and the ultrastructure of the LA cells was observed under scanning electron microscope .The expression of Cav 1.2 in LA tissue of the 2 groups was detected by Western blot . RESULTS:Compared with young LA group , dv/dtmax and plateau potential were significantly decreased , APD30 and APD50 were shortened, and APD90 was notably prolongated in aged LA group (P<0.01).The inducibility or duration of atrial arrhythmias was severely increased or prolongated in aged LA group (P<0.01).With voltage clamp model, the den-sity of ICa,L in aged LA group was significantly decreased , and current-voltage curve was notably moved upward compared with young LA group.When the clamp potential was +20 mV, the density of ICa,L was notably modified from (11.72 ± 1.39) pA/pF in young LA group to (6.08 ±0.98) pA/pF in aged LA group ( P<0.01).Compared with young LA group, the protein level of collagen was significantly increased (P<0.01), and the arrange of atrial myocytes was irregular in LA of rabbits in aged LA group .The atrial myocytes of the LA wall in aged LA group exhibited abnormal ultrastructural alterations , such as karyopyknosis , irregular and swelling mitochondria with the presence of vacuoles , and mild and severe sarcomere degeneration .Compared with those in LA tissues of young rabbits , the expression levels of Cav 1.2 in the LA tis-sues of aged rabbits were severely reduced (P<0.01), and had a significant positive correlation with the reduction of ICa,L (r=0.83, P<0.01).CONCLUSION:The pathophysiological characteristics of aged LA are significantly altered , and might contribute to vulnerability and susceptibility of occurrence of atrial fibillation in aged rabbits .The mechanisms might completely attribute to the notable reduction of ICa,L , abnormal alterations of ultrastructures and obvious decrease in the ex-pression of Cav1.2 in the aged LA of aged rabbits .

Objective To study the effectsof rehabilitative training on neural function and the expression of glial fibrillary acidic protein (GFAP) and ionized calcium binding adaptor molecule-1 (Iba-1) in rats after traumatic brain injury.Methods A left hemisphere traumatic brain injury model was established in ninety Sprague-Dawley rats.They were then randomly divided into a rehabilitation training group,an immobilization group and a free-running group,with 30 rats in each group.Another thirty rats received sham injury as the shamoperated group.Beginning 4 days post-operation the rats of the rehabilitation training group were given balancing,rotating and walking exercises three times daily,15 min/time,6 d/week.The immobilization group was fixed in mesh cages.The rats of the free-running and sham-operated groups were reared in normal cages without any special training exercise.The rats of all 4 groups were given neural and motor function tests to assess the functional outcome.Immunofluorescence staining was used to evaluate the expressions of GFAP (the marker of astrocytes) and Iba-1 (the marker of microglia) in the cortex close to the iujured region at 3 days,1 week,2 weeks,3 weeks and 4 weeks after injury.Results The average neural and motor function test scores in the rehabilitation training group were significantly better than those in the immobilization and free-running groups at the 2nd week and thereafter.The average scores in the free-running group were significantly better than those in the immobilization group at the 4th week after injury.The immunofluorescence staining showed that the expression of GFAP was lowest in the rehabilitation group at the 2nd week and thereafter.Iba-1 expression was significantly lower only at the 3rd week and beyond.Conclusion Rehabilitative training can improve nerve function recovery in rats after traumatic brain injury,and the functional enhancement may be partially attributed to the downregulation of activated astrocytes and microglia.

Objective To investigate the impact of transvaginal natural orifice transluminal endo-scopic surgery(NOTES)-assisted laparoscopic nephrectomy on female sexual function and quality of life . Methods This was a prospective study on the change of female sexual function and quality of life of female patients who underwent transvaginal NOTES-assisted laparoscopic nephrectomy from May .2011 to Nov. 2012.A total of 42 cases were included in this study (28 of them with severe hydronephrosis , non-functio-ning kidney , 11 with pyelonephrosis , 1 with renal tuberculosis , 1 with duplex kidney complicated with hy-dronephrosis, and 1 with renal angiomyolipoma ).The mean age was 36.9±5.3 (26-45) years, and the mean body mass index was 21.7±2.6 (14.7 to 27.1) kg/m2.Twenty-four cases were operated on the left side, 18 cases on the right .The female sexual function and quality of life were assessed before and 4 months, 7 months and 1 year after surgery using the Female Sexual Function Index (FSFI) questionnaire and the MOS 36-item Short-Form Health Survey (SF-36), respectively. Results The mean FSFI of 42 cases preoperatively and 4 months, 7 months and 1 yr postoperatively were 27.74 ±4.34, 27.19 ±4.49, 28.54±4.23, and 28.68 ±4.19, respectively.There was no statistically significant difference among them (F=1.111, P=0.346).Compared with that of preoperation , the physical function, vitality, metal health, body pain, and general health of the patients were improved , but the role-physical, role-emotion and social function were not improved at postoperative month 4 and month 7 (P<0.05).Each item of SF-36 was im-proved after postoperative 1 year ( P<0.05) . Conclusions Transvaginal NOTES-assisted laparoscopic ne-phrectomy does not cause negative effect on the female sexual function .The quality of life can be improved after operation .The physical function is improved at early stage , and the psychological function as well .

This study aimed to identify the related substances of lorazepam tablets by liquid chromatography mass spectrometry (LC-MS). To separate the related substances of lorazepam tablets, gradient elution was performed using acetonitrile and 0.1% acetic acid -20 mmol/L of ammonium acetate as mobile phase on Inert Sustain C18 (250 mm × 4.6 mm, 5 μm).The accurate mass and elemental composition of the parent ions and their product ions of related substances were determined by electrospray-ionization quadrupole time-of-flight high resolution mass spectrometry (ESI-Q-TOF/MS).The structures of the related substances were identified by spectral analysis. Under the established analytical condition, lorazepam and its related substances were adequately separated, and 22 major related substances with content greater than 0.1% were detected and identified by hyphenated techniques in lorazepam tablets and their stressed samples.Among them, 5 were the impurities listed in the USP or EP, and the others were unknown related substances identified for the first time in this paper.The LC-MS technique can effectively separate and identify the related substances of lorazepam tablets, which provides some reference for quality control.

Cinnabaris(α-HgS) is a mineral traditional Chinese material medica, as a tranquilizer and sedative, which is widely used in combination with herbs for the treatment of children high fever and convulsion.However, a large amount of mercury in Cinnabaris poses a potential risk to the immature central nervous system of children and probably causes severe memory disorders.Inthisstudy,three groups of juvenile rats were given low, medium, and high doses of Cinnabaris by oral gavage once a day for 14 continuous weeks, respectively.The blood mercury concentrations of the rats at different growth phases were monitored by atomic fluorescence spectrometry.The brain structural and functional changes related to the memory functions were investigated through HE staining and Morris water-maze test. Correlation analysis was conducted to clarify the dose- mercury exposure-toxic effect relationship of Cinnabaris and memory disorders.It was found thatthe blood mercury levels increased in both time- and dose-dependent manner.After the 14-week continuous administration of Cinnabaris, the pathological lesions in hippocampal neurons of rats in the high dose group were observed including pyknosis and disordered cell arrangement.In the Morris water-maze test, compared with the control group, rats in the high dose group exhibited the significantly prolonged latency to find the platform and the target quadrant, and the time spent in the target quadrant was obviously shortened. Thus, the significant correlations were established between Cinnabaris dose and mercury exposure,mercury exposure and memory disorders, respectively. In conclusion, the long-term and overdose administration of Cinnabaris in juvenile rats can increase the in-vivo mercury level, destroy the normal hippocampal morphological structure, and lead to memory disorders. This study provided scientific references for the potential mercury poisoning risks pharmacovigilance of Cinnabaris-containing paediatric formulations.

To establish a method for the determination of formaldehyde and glyoxal simultaneously in varenicline tartrate active pharmaceutical ingredient (API) and its intermediate, formaldehyde and glyoxal were derivatized by 2, 4-dinitrophenylhydrazine (2,4-DNPH) to improve the HPLC retention and UV detection sensitivity. Separation was performed on a C8 (150 mm × 4.6 mm, 5 μm) column by linear gradient elution using acetonitrile and water as the mobile phase; the detective wavelength was set at 380 nm.Formaldehyde and glyoxal were quantitatively determined by an external reference method.Linear calibration was established for both formaldehyde and glyoxal in the range from 0.094 to 1.88 μg/mL.The detection and the quantification limits were 0.047 μg/mL (19 μg/g) and 0.094 μg/mL (38 μg/g), respectively.The recoveries were (95.0±1.1)% and (99.4 ± 2.6)% for formaldehyde and glyoxal, respectively.This method has been fully validated to be applicable to quantitative analysis of trace amount of formaldehyde and glyoxal in varenicline tartrate API and its intermediate.Test results demonstrated that the contents of both formaldehyde and glyoxal met the permitted daily exposure (PDE) limits for the finished products of varenicline tartrate API as well as its intermediate, though the glyoxal contents in the crude intermediates were likely to exceed the limit.The established method is valuable for the manufacturing process and quality control of varenicline tartrate.

Objective To describe the initial clinical experience of transvaginal NOTES-assisted laparoscopic nephrectomy. Methods From May to June 2010, 4 female patients with non-functioning kidney and 1 female patient with renal tuberculosis underwent transvaginal NOTES-assisted laparoscopic nephrectomy. The median age was 41 years (range 36 to 63). Three lesions were on the right side, and 2 on the left. After induction of general anesthesia, the patients were positioned in lithotomy with ipsilateral lumbar at 60° angle to the operating table. A 5 mm Trocar and a 10 mm Trocar were placed through the umbilicus, and a 10 mm Trocar for 30° laparoscope was placed through the posterior vaginal fornix under direct vision. The kidneys were put into a home-made bag and removed via the incision of posterior vaginal fornix after there were completely resected. Results The 5 procedures were successfully finished. The median operative time was 190 (range 150 to 260)min. The median estimated blood loss was 185 (range 150 to 210) ml. There were no intra-operative or postoperative complications. The patients resumed ambulation 1 day after surgery, and resumed nutrition 2 days after surgery. The drainages were removed on postoperactive day 3. The patients were discharged on postoperative day 7. The incision in vagina healed well. Conclusions Transvaginal NOTES-assisted laparoscopic nephrectomy is feasible and reproducible. This novel technique may provide additional benefits related to postoperative pain, recovery, and cosmetic results even when compared to today's minimally invasive procedures.

Objective:To investigate the effect of astragaloside IV (AS-IV) regulating the signal axis of stromal cell-derived factor-1α (SDF-1α)/CXC chemokine receptor 4 (CXCR4) on the mobilization of bone marrow endothelial progenitor cells (EPCs) to peripheral blood in diabetes skin ulcer (DSU) rats.Methods:Twenty four SPF grade male Sprague Dawley (SD) rats were selected to make the model of type 2 diabetes rats by intraperitoneal injection of 30 mg/kg 1% (plastid ratio) streptozotocin, and then round full-thickness skin with a diameter of 2 cm was cut on both sides of the waist and back to make the skin ulcer model of diabetes rats. After that, they were randomly divided into AS-IV group (50 mg/kg AS-IV), blocker group (50 mg/kg AS-IV+ 5 mg/kg AMD3100) and model group. At the same time, a blank group ( n=8) was set up, The drug was administered via intraperitoneal injection, and the model group and blank group were treated with 0.9% NaCl of equal volume. On the 10th day, peripheral blood, femoral bone marrow, and wound neovascularization tissues of rats were collected. The number of EPCs in peripheral blood of each group of rats was measured by flow cytometry, and the protein expression of SDF-1α and CXCR4 in peripheral blood, femoral bone marrow, and wound neovascularization tissues of rats was detected by enzyme-linked immunosorbent assay (ELISA); At the same time, the wound healing rates of each group were tested. Results:On the 10th and 21st day after modeling, the wound healing rate of each group of rats was compared. The blank group healed the fastest, while the model group healed the slowest. The AS-IV group had better healing than the model group and the blocker group, and the difference was statistically significant (all P<0.05). On the 10th day after modeling, the positive rates of peripheral blood EPCs in the white group, AS-IV group, and blocker group were significantly higher than those in the model group (all P<0.05), while the positive rates of peripheral blood EPCs in the blocker group were significantly lower than those in the AS-IV group (all P<0.05). On the 10th day after modeling, the protein expression of SDF-1α and CXCR4 in the wound, serum, and bone marrow of the model group was the lowest, while the protein expression in the blank group was the highest (all P<0.05). The protein expression of SDF-1α and CXCR4 in the wound, serum, bone marrow of the AS-IV group was significantly higher than that of the blocker group and model group, and the difference was statistically significant (all P<0.05). Conclusions:Astragaloside IV can promote the mobilization and migration of endothelial progenitor cells from bone marrow to peripheral blood in diabetes ulcer rats by regulating SDF-1α/CXCR4 signal axis, and can participate in angiogenesis of diabetes ulcer wounds as seed cells to promote the healing of diabetes skin ulcers.

Obesity related glomerulopathy (ORG), as a disease with an increasing incidence of metabolic kidney injury, has become a new hot spot in today's society. A variety of factors are involved in the occurrence and development of ORG. Among them, the ectopic deposition of kidney lipids is not only a significant feature of ORG, but also a key link to promote the progress of ORG. This article reviews the related mechanisms of lipid deposition in ORG and the treatment of ORG with lipid deposition as the target.