Protective Mechanism of Danggui Shaoyaosan on Podocytes of Nephrotic Syndrome Rats Based on AngⅡ-TRPC6 Pathway / 中国实验方剂学杂志

Objective:To explore the protective effect and the mechanism of Danggui Shaoyaosan（DSS） on angiotensin Ⅱ （AngⅡ）/transient receptor potential cation channel 6 （TRPC6） pathway in nephrotic syndrome （NS） rats. Method:In animal experiments， doxorubicin （4 mg·kg^-1 for the 1^st week and 2 mg·kg^-1 for the 2^nd week） was injected twice to the tail vein of rats to induce NS model in 160 rats， which were then randomly divided into model group （normal saline）， losartan group （30 mg·kg^-1·d^-1）， and low-（4.3 g·kg^-1·d^-1）， medium-（8.6 g·kg^-1·d^-1）， and high-dose （17.2 g·kg^-1·d^-1） DSS groups. Besides， a normal group was also set. After intervention for four weeks， ultrastructure changes of the kidney were identified by transmission electron microscopy （TEM）. The 24-hour urine protein was detected by kits. Radioimmunoassay was used to detect the content of AngⅡ and Calcineurin （CaN） in plasma. Western blot was used to detect the protein expression of TRPC6， angiotensin Ⅱ type 1 receptor （AT1R）， podocyte slit diaphragm-specific protein （Nephrin）， and cysteine-aspartic acid protease-3 （Caspase-3） in the renal cortex. Immunohistochemistry was used to detect the expression of TRPC6 and AT1R in the slit diaphragm. In cell experiments， AngⅡ stimulated MPC5 podocytes. The cells were randomly divided into a normal group， an AngⅡ group， an AngⅡ+SAR7334 （TRPC6-specific inhibitor） group， an AngⅡ+5%DSS group， an AngⅡ+10%DSS group， and an AngⅡ+15%DSS group. Western blot was used to detect the protein expression of TRPC6， AT1R， Nephrin， and Caspase-3 in podocytes. Result:Compared with the normal group， the model group showed increased 24-hour urine protein content （<italic>P</italic><0.01） and AngⅡ and CaN in plasma （<italic>P</italic><0.01）， incomplete glomerular structure， the extensive fusion of podocyte process with elevated fusion rate （<italic>P</italic><0.01）， increased expression distribution of AT1R and TRPC6 in the renal cortex， and up-regulated protein expression of AT1R， TRPC6， and Caspase-3 in renal tissues （<italic>P</italic><0.01）， and reduced Nephrin protein expression （<italic>P</italic><0.01）. Compared with model group， the losartan group and the high-dose DSS group exhibited decreased 24-hour urine protein content （<italic>P</italic><0.01） and the content of AngⅡ and CaN in plasma （<italic>P</italic><0.01）， improved glomerular structure， reduced fusion rate of podocyte process （<italic>P</italic><0.01）， diminished expression distribution of TRPC6 and AT1R in the renal cortex， declining protein expression of AT1R， TRPC6 and Caspase-3 in renal tissues （<italic>P</italic><0.01）， and elevated Nephrin protein expression （<italic>P</italic><0.01）. Additionally， compared with the normal podocytes， AngⅡ-stimulated podocytes showed increased protein expression of AT1R， TRPC6 and Caspase-3 （<italic>P</italic><0.01）， and decreased expression of Nephrin （<italic>P</italic><0.01）. Compared with the AngⅡ group， the AngⅡ+SAR7334 group displayed reduced protein expression of AT1R， TRPC6， and Caspase-3 （<italic>P</italic><0.01） and increased protein expression of Nephrin （<italic>P</italic><0.01）. Conclusion:DSS can improve the pathological characteristics of NS presumedly by inhibiting the interaction between AngⅡ and TRPC6 in podocytes and improving the structural integrity of podocytes to repair the damage of glomerular molecular barrier and slow down the progression of NS-induced proteinuria.

Evaluation of Rat Model with Spleen Deficiency and Dampness Stagnancy Based on Bioelectrical Impedance / 中国实验方剂学杂志

Objective: To evaluate the model with spleen deficiency and dampness stagnancy by bioelectrical impedance analysis (BIA) and traditional indicators. Method: The forty rats were divided into blank group and model group, with 20 rats in each group. The rats in the blank group were fed with normal feed, the rats in model group were prepared with the spleen deficiency and dampness stagnancy model for 14 days. Observe the general condition of the rats, measure the water content of the feces in the dry method, measure the water load index by weighing method, and detect the urinary D-xylose excretion total protein (TP), albumin (Alb) content, by enzyme-linked immunosorbent assay (ELISA). Western blot analysis of renal aquaporin 1 (AQP1) content, and the use of experimental animal body composition analyzer to determine the total water content (TBW), extracellular fluid (ECF), intracellular fluid (ICF), fat mass (FM), free fat mass (FFM) and body mass bioelectrical impedance index such as body mass index (BMI). Result: Compared with blank group, the rats in model group lost weight, gradually loose stools occasionally, the anus temperature was basically unchanged, body mass, D-xylose excretion, water load index, TP and Alb content decreased (PPPConclusion: Rats with spleen deficiency and dampness stagnancy induced a combination of factors such as diet and excessive fatigue. The bioelectrical impedance method can be more intuitive and comprehensive.

Effect of Danggui Shaoyao San Drug-containing Serum on Expression of p-MLCⅡ and MLCⅡ Protein in HSC-T6 Cells Induced by ET-1 / 中国实验方剂学杂志

Objective:To investigate the effect of endothelin-1 (ET-1) on the expression of phosphorylated myosin light chain Ⅱ(p-MLCⅡ)and myosin light chain Ⅱ(MLCⅡ)protein in rat hepatic stellate cells HSC-T6 and explore the intervention effect of Danggui Shaoyao San(DSS)drug-containing serum. Method:After HSC-T6 cells were seeded, DMEM and blank rat serum with final concentrations of 2.5%, 5%, 10%, 15% and 20% were added to each well. The viability of HSC-T6 cells was determined by methyl thiazolyl tetrazolium(MTT) assay to screen the suitable serum concentration range. The cells were divided into blank serum control group (5%, 10%, 15%) and DSS drug-containing serum group (5%, 10%, 15%). ELISA was used to detect the content of ET-1 in cell culture supernatant under basic state. The cells were divided into blank serum control group (10%), DSS drug-containing serum low (5%), medium (10%) and high dose (15%) groups. Real-time fluorescent quantitative polymerase chain reaction (Real-time PCR) was used to detect the level of ET-1 mRNA in cell culture supernatant under basic state. The cells were divided into blank serum control group (10%), model group (10%), DSS drug-containing serum low (5%), medium (10%), high dose (15%) groups and Y-27632 inhibitor group (100 μmol·L-1). Except the blank serum control group, the other groups all received 10 nmol·L-1 ET-1 to induce HSC-T6 cells. Western blot was used to detect the expression of p-MLCⅡ and MLCⅡ in HSC-T6 cells induced by ET-1. Result:Serum concentrations of 5%, 10% and 15% were used as drug-containing serum concentrations. As compared with the blank serum control group, the DSS drug-containing serum group significantly reduced the relative content of ET-1 and ET-1 mRNA in the basic state (PPPPPConclusion:DSS drug-containing serum may down-regulate the expression of p-MLCⅡ and MLCⅡ by down-regulating the content of ET-1 and inhibiting the autocrine of ET-1.

A preliminary study on origin of ligustrazine in Chuanxiong Rhizoma based on endogenetic Bacillus subtilis / 中国中药杂志

Ligustrazine is an important active ingredient of the traditional Chinese medicine Chuanxiong Rhizoma, but its content is a controversial topic. The endophytes of medicinal plants have the ability to produce the same active substances as the host, so this report focused on the endophytic Bacillus subtilis, to study the origin of ligustrazine in Chuanxiong Rhizoma preliminarily by inoculating the isolated endophytic B. subtilis to the Chuanxiong Rhizoma medium for solid state fermentation. Tissue grinding method was used to isolate the endogenetic B. subtilis. The morphological features, conventional physiological and biochemical reactions and 16S rRNA molecular techniques were combined to identify the endogenetic strains. Then, the strains that grew well in the medicinal matrix of Chuanxiong Rhizoma were screened out for further fermentation studies. The solid-state fermentation was performed at 37 °C for 30 d using Chuanxiong Rhizoma fermentation medium (40 g Chuanxiong Rhizoma powder, 100 mL sterile water, 121 °C, sterilization for 25 minutes). UPLC was used to detect the contents of ligustrazine, acetoin in the Chuanxiong Rhizoma fermentation medium and Chuanxiong Rhizoma. All the five strains were Gram-positive and had spores. Phylogenetic analysis of the 16S rRNA sequence showed that the endophytes were B. subtilis. The results of UPLC showed that ligustrazine was detected in the Chuanxiong Rhizoma fermentation medium inoculated with endogenetic B. subtilis LB3, LB3-2-1, LB4, LB5 and LB6-2, while not detected neither in blank Chuanxiong Rhizoma fermentation medium nor in Chuanxiong Rhizoma. This study showed that the endogenetic B. subtilis of Ligusticum chuanxiong Hort. can make use of Chuanxiong Rhizoma fermentation medium to produce ligustrazine. Endogenetic B. subtilis has a certain correlation with the accumulation of ligustrazine in Rhizoma Chuanxiong. We speculate that the ligustrazine may be derived from the catabolism of endogenetic B. subtilis in Ligusticum chuanxiong.

Clinical study on the changes of the tumor target volume and organs at risk in helical tomotherapy for nasopharyngeal carcinoma / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Helical tomotherapy (HT) is a new image-guided intensity-modulated radiation therapy (IMRT). The aim of this study was to evaluate the changes in the target volume and organs at risk (OARs) of patients with nasopharyngeal carcinoma (NPC) during helical tomotherapy.</p><p><b>METHODS</b>Forty-three patients with NPC and treated via HT from March 2008 to January 2010 were reviewed retrospectively. Repeated CT scanning and plan adaptation were conducted at the 20th fraction during radiotherapy. The volumetric differences between the two scans were evaluated for nasopharyngeal tumor and retro-pharyngeal lymph nodes (GTVnx), neck lymph nodes (GTVnd), and parotid glands, as well as the axial diameter of the head.</p><p><b>RESULTS</b>The median interval between the two scans was 25 days (23 - 28 days). The volumetric decrease in GTVnx was 30.1% (median, 29.8%) and in GTVnd 41.6% (median, 45.9%). The variation in the GTVnd volume was correlated with the weight loss of the patient. The volume of the left parotid gland decreased by 35.5% (median, 33.4%) and of the right parotid glands decreased by 36.8% (median, 33.5%). The axial diameter of the head decreased by 9.39% (median, 9.1%).</p><p><b>CONCLUSIONS</b>The target volume and OARs of patients with NPC varied considerably during HT. These changes may have potential dosimetric effects on the target volume and/or OARs and influence the clinical outcome. Repeated CT scanning and replanning during the HT for NPC patients with a large target volume or an obvious weight loss are recommended.</p>

Dosimetric comparison between helical tomotherapy and intensity-modulated radiation therapy plans for non-small cell lung cancer / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Helical tomotherapy (HT) is a new image-guided intensity-modulated radiation therapy (IMRT) technique. It is reported that HT plan for non-small-cell lung cancer (NSCLC) can give better dose uniformity, dose gradients, and protection for the lung than IMRT plan. We compared the dosimetric characteristics of HT for NSCLC with those of conventional IMRT to observe the superiority of HT.</p><p><b>METHODS</b>There was a comparative case series comprising 10 patients with NSCLC. Computed tomographic (CT) images of delineated targets were transferred to the PrecisePlan planning system (IMRT) and Tomo planning system (HT). The prescription doses were 70 Gy/33F for the gross tumor volume (GTV) and the visible lymph nodes (GTVnd), and 60 Gy/33F for the clinical target volume (CTV) and the clinical target volume of the visible lymph nodes (CTVnd). The dose restrictions for organs at risk were as follows: the maximum dose to spinal cord ≤ 45 Gy, V20 to the total lungs < 30%, V50 to the heart < 50%, and V55 to the esophagus < 50%. Both plans were evaluated by means of the dose coverage of the targets, dose-volume histograms (DVHs), and other dosimetric indices.</p><p><b>RESULTS</b>The dose coverage, conformity, and homogeneity of the targets' volumes were found to be satisfactory in both plans, but the homogeneity of the HT plan was better than that of IMRT. The high-dose radiation volume (V20-V30) to the lung and the mean lung dose (MLD) decreased (P < 0.05), but the low-dose radiation volume (V5-V10) increased slightly in the HT plan (P > 0.05). The maximum doses to the spinal cord, heart, esophagus and trachea in the HT plan were lower than those in the IMRT plan, but the differences were not statistically significant.</p><p><b>CONCLUSIONS</b>The HT plan provids better dose uniformity, dose gradients, and protection for the organs at risk. It can reduce the high-dose radiation volume for lung and the MLD, but may deliver a larger lung volume of low-dose radiation.</p>

Clinical observation on treatment of post-craniocerebral traumatic mental disorder by integrative medicine / 中国结合医学杂志

<p><b>OBJECTIVE</b>To observe the clinical efficacy of integrative medical therapy in treating post-craniocerebral traumatic mental disorder (PCT-MD).</p><p><b>METHODS</b>Sixty patients with confirmed diagnosis of PCT-MD were randomly assigned to the treated group and the control group equally. All were treated by conventional comprehensive Western medicine, but to patients in the treated group, modified Xufu Zhuyu Decoction (XFZY) was additionally given and the therapeutic course for both groups was 20 days. Changes in mental symptoms were observed and recorded on the 10th and 20th day and clinical efficacy as well as cranial CT image was estimated after termination of the treatment.</p><p><b>RESULTS</b>The clinical effective rate in the treated group and the control group was 96.67% and 83.30% respectively. Comparison between them showed significant difference (P<0.05). Significant differences were also shown in the comparisons between the two groups in improving mental symptoms, either on the 10th or on the 20th day (P<0.05 and P<0.01 respectively), and in post-treatment cranial CT image (P<0.05).</p><p><b>CONCLUSION</b>Better efficacy could be obtained by integrative medical therapy in treating PCT-MD.</p>

Verification of accuracy of multileaf collimator leaf position using a two-dimensional ion chamber array / 中华放射肿瘤学杂志

Objective To design a new method to verify the position of multileaf collimator(MLC)leaf using a two-dimensional ion chamber array(2D-array).Methods 2D-array of PTW T10018 Seven29~(TM) was used to calibrate the accuracy of MLC leaf position of Elekta Precise accelerator.The edge function of the leaf position of MLC was measured and used as the reference value.The precision of MLC leaf was then evaluated through comparing the measured and reference values.Results The accuracy of MLC leaf position was found within?0.1 mm.Conclusion This method of verifying the accuracy of multileaf collimator leaf position is easy,simple and reliable