Effects of Rehmanniae Radix and Rehmanniae Radix Praeparata on proteomics and autophagy in mice with type 2 diabetes mellitus induced by high-fat diet coupled with streptozotocin / 中国中药杂志

To compare the pancreatic proteomics and autophagy between Rehmanniae Radix-and Rehmanniae Radix Praeparata-treated mice with type 2 diabetes mellitus(T2DM). The T2DM mouse model was established by high-fat diet coupled with streptozotocin(STZ, intraperitoneal injection, 100 mg·kg~(-1), once a day for three consecutive days). The mice were then randomly assigned into a control group, low-(5 g·kg~(-1)) and high-dose(15 g·kg~(-1)) Rehmanniae Radix groups, low-(150 mg·kg~(-1)) and high-dose(300 mg·kg~(-1)) catalpol groups, low-(5 g·kg~(-1)) and high-dose(15 g·kg~(-1)) Rehmanniae Radix Praeparata groups, low-(150 mg·kg~(-1)) and high-dose(300 mg·kg~(-1)) 5-hydroxymethyl furfuraldehyde(5-HMF) groups, and a metformin(250 mg·kg~(-1)) group. In addition, a normal group was also set and each group included 8 mice. The pancreas was collected after four weeks of administration and proteomics tools were employed to study the effects of Rehmanniae Radix and Rehmanniae Radix Praeparata on protein expression in the pancreas of T2DM mice. The expression levels of proteins involved in autophagy, inflammation, and oxidative stress response in the pancreatic tissues of T2DM mice were determined by western blotting, immunohistochemical assay, and transmission electron microscopy. The results showed that the differential proteins between the model group and Rehmanniae Radix/Rehmanniae Radix Prae-parata group were enriched in 7 KEGG pathways, such as autophagy-animal, which indicated that the 7 pathways may be associated with T2DM. Compared with the control group, drug administration significantly up-regulated the expression levels of beclin1 and phosphorylated mammalian target of rapamycin(p-mTOR)/mTOR and down-regulated those of the inflammation indicators, Toll-like receptor-4(TLR4) and Nod-like receptor protein 3(NLRP3), in the pancreas of T2DM mice, and Rehmanniae Radix showed better performance. In addition, the expression levels of inducible nitric oxide synthase(iNOS), nuclear factor erythroid 2-related factor 2(Nrf2), and heine oxygenase-1(HO-1) in the pancreas of T2DM mice were down-regulated after drug administration, and Rehmanniae Radix Praeparata demonstrated better performance. The results indicate that both Rehmanniae Radix and Rehmanniae Radix Praeparata can alleviate the inflammatory symptoms, reduce oxidative stress response, and increase the autophagy level in the pancreas of T2DM mice, while they exert the effect on different autophagy pathways.

Rehmanniae Radix and Rehmanniae Radix Praeparata improve diabetes induced by high-fat diet coupled with streptozotocin in mice through AMPK-mediated NF-κB/NLRP3 signaling pathway / 中国中药杂志

This study investigated the differential mechanisms of Rehmanniae Radix and Rehmanniae Radix Praeparata in improving diabetes in mice through AMPK-mediated NF-κB/NLRP3 signaling pathway. The diabetic mouse model was established with high-fat diet coupled with streptozotocin(STZ, intraperitoneal injection, 100 mg·kg~(-1), once a day for three consecutive days), after which the mice were randomly divided into model group, low-dose(5 g·kg~(-1)) and high-dose(15 g·kg~(-1)) Rehmanniae Radix groups, low-dose(5 g·kg~(-1)) and high-dose(15 g·kg~(-1)) Rehmanniae Radix Praeparata groups, catalpol group(250 mg·kg~(-1)), 5-hydroxymethylfurfural(5-HMF) group(250 mg·kg~(-1)), metformin group(250 mg·kg~(-1)), with the normal group also set. The organ indexes of heart,liver, spleen, lung, kidney and pancreas were calculated after four weeks of administration. The pathological changes and fibrosis of pancreas, kidney and liver in mice were observed by hematoxylin-eosin(HE) staining and Masson staining. Western blot was used to determine the expression levels of Toll-like receptor-4(TLR4), nuclear factor-κB(NF-κB), Nod-like receptor protein 3(NLRP3),interleukin-1β(IL-1β), adenosine monophosphate-activated protein kinase(AMPK), phosphorylated AMPK(p-AMPK) in the pancreas, kidney and liver of mice. Compared with the model group, the administration groups witnessed significant decrease in the liver,spleen, kidney, pancreas and fat indexes of diabetic mice, and there was no significant difference in heart and lung indexes. The pathological states and fibrosis of pancreatic, kidney and liver tissues were significantly improved after administration. Additionally, the expression levels of TLR4, NF-κB and NLRP3 in pancreas, kidney and liver of diabetic mice were significantly lowered. The expression levels of p-AMPK/AMPK were enhanced significantly in kidney and liver of mice in Rehmanniae Radix group while in pancreas, kidney and liver in Rehmanniae Radix Praeparata group. This suggests that Rehmanniae Radix and Rehmanniae Radix Praeparata differ in the mechanism of regulating energy metabolism of multiple organs and thereby exerting anti-inflammatory effects to alleviate symptoms of diabetic mice.

Study on the Synthesis of Shikimic Acid Derivatives and Their Reversal Effects on Paclitaxel-resistant Human Breast Cancer Cells / 中国药房

OBJECTIVE：To struc turally modify shikimic acid ，and to investigate the reversal effects of its derivatives on paclitaxel-resistant human breast cancer cells MCF- 7/PTX. METHODS ：Using shikimic acid as the lead structure ，1-position carboxyl group was structurally modified to synthesize a series of shikimic acid derivatives through esterification ，amidation， hydrogenation and reduction ，etc. Using non-drug resistant cells MCF- 7 as reference ，MTT assay was used to screen derivatives with inhibitory activity as well as half-inhibitory concentration （IC50）and reversal index （RI）of derivatives to MCF- 7/PTX. With the drug resistance-related transgelin 2 as the target ，the molecular docking of the active derivatives with the drug resistance-related protein was carried out by using Glide 1.0 computer-aided design software. RESULTS ：Totally 15 derivatives were obtained （T1-T15）， of which T 4-T15 were obtained for the first time. MTT assay showed that （3R， 4S， 5R） -N-benzyl-3， 4， 5-trihydroxy-1-cyclohexene-1-formamide（T7），（3R，4S，5R）-N-（3，4，5-trihydroxy-1-cyclohexenylmethyl）-benzylamine（T14）， （3R，4S，5R）-3，4-O-isopropyl-5-O-acetyl-1-cyclohexene-1-methyl formate （T15）inhibited MCF- 7 and MCF- 7/PTX cells to a certain extent ；IC50 values of T 7，T14 and T 15 combined with pacliaxel to MCF- 7/PTX cells were significantly lower than that in negative control （Paclitaxel alone ）group（P＜0.05）. RIs of T 14 and T 15 were higher ，and RIs of the highest dose were 8.8 and 9.3， which were equivalent to positive control verapamil （10.8）. Th e results of molecular docking showed that the hydroxyl groups at positions 3，4 of T 7 could form multiple hydrogen bonds with ； Arg625 and Asp 627 in the catalytic region of transgelin 2. In addition to the hydrogen bond mentioned above at T 7，the mail：batistuta28@126.com secondary amine side chain at position 1 of T 14 could also form hydrogen bond with Glu 657 of transgelin 2. When the hydroxyl group on the T 15 mother nucleus was derived from the donor group ，the binding of the hydroxyl group to transgelin 2 was closer and the inhibition was enhanced. CONCLUSIONS ： The derivatives T 7，T14 and T 15 have certain reverse activity to paclitaxel-resistant human breast cancer cells. The polyhydroxy structure of the mother nucleus is the main structural region of the hydrogen bond between shikimic acid and its derivatives and transgelin 2. The derivation of its power supply group or the introduction of secondary amines and hydrophobic groups into the 1-carboxyl group of shikimic acid is benifit for enhancing the drug resistance reversal effect of derivative .

Associations between normal serum levels of uric acid and knee symptoms, joint structures in patients with knee osteoarthritis / 中华疾病控制杂志

Objective To investigate the associations between normal serum uric acid (UA) levels and knee symptoms and structural changes in patients with knee osteoarthritis (OA).Methods A total of 205 patients with knee OA were enrolled. According to the inclusion and exclusion criteria, 122 patients with OA were included in the study. The general condition of patients were investingated. The Western Ontario and McMaster Universities Osteoarthritis Index (WOMAC) score was used to assess severity of the knee joint symptoms. The Kellgren-Lawrence (K-L) grading system was used to assess radiological severity of the knee joint. Osirix software was used to assess changes of knee structure in magnetic resonance imaging (MRI). Peroxidase method was used to measure the serum levels of uric acid. Patients with normal range of uric acid were included in the study.Results The patients with lower levels of UA (split at the median level, 263 μmol/L) had a higher proportion of women, lower height, weight and lower proportions of K-L grade≥2. After adjustment for sex, age and body mass index, serum UA was negatively associated with total WOMAC scores (β=-16.15, P=0.018), WOMAC pain (β=-3.15, P=0.037), and WOMAC stiffness (β=-1.65, P=0.025). However, there were no significant associations between serum UA levels and knee joint structure changes.Conclusions In OA patients with normal serum UA, serum UA is negatively associated with the severity of joint symptoms, suggesting that UA within normal range may play a protective role in OA symptoms.

Effect of baicalein on pancreas insulin secretion in rats and its mechanism / 中国药理学通报

Aim To investigate the effect of baicalein on insulin secretion from rat islets and the underlying mechanism. Methods Pancreatic islets were obtained from the pancreas of male Wistar rats by collagenase P digestion and histopaque-1077 density gradient separa-tion. Single islet cells were dispersed from pancreatic islets by Dispase II digestion. Insulin secretion experi-ment was applied to observe insulin release after baica-lein stimulation. To study the potential mechanism, calcium imaging technique and patch-clamp experiment were applied to measure intracellular Ca2+ concentra-tion and voltage-dependent potassium channel currents (Kv). Results In 16. 7mmol·L-1 glucose, baica-lein accelerated insulin secretion in a dose-dependent manner. Baicalein promoted the intracellular Ca2+ con-centration. The patch-clamp experiment showed that baicalein inhibited Kv current in a dose-dependent manner. Conclusion Baicalein can increase the in-tracellular Ca2+ concentration by inhibiting Kv chan-nels and eventually promoting insulin secretion.

Ear reconstruction after rupture of postauricular expanded flap / 中华整形外科杂志

<p><b>OBJECTIVE</b>To summarize the principles of ear reconstruction after rupture of postauricular expanded flap.</p><p><b>METHODS</b>The rupture of the postauricular expanded flap was graded as I-IV degree. For flap rupture of I and II degree, the ear reconstruction was performed immediately with autogenous cartilage framework. For flap rupture of III degree, topical and systemic treatment should be done to relieve the redness and swelling. Then the ear reconstruction was performed. For flap rupture of IV degree, the expanded postauricular flap was smoothened. Ear reconstruction was performed 3 months later.</p><p><b>RESULTS</b>There were 67 cases with flap rupture, including 43 cases as I and II degree, 17 cases as III degree and 7 cases as IV degree. Ear reconstruction with autogenous cartilage framework was performed in 43 cases, with Medpor framework in 17 cases. Flap smoothing and secondary ear reconstruction with autogenous cartilage framework was performed in 7 cases. The results were satisfactory with complications in 4 cases (7%), which resolved after treatment. Primary healing was achieved in all the patients.</p><p><b>CONCLUSIONS</b>Good result could be achieved in ear reconstruction, even after flap rupture. Proper treatment should be adopted according to the rupture degree.</p>

Establishment of genotyping method for fetal ABO group from pregnant maternal peripheral blood / 中国实验血液学杂志

This study was aimed to establish a genotyping method to detect ABO group gene of fetus from peripheral blood of pregnant women for prenatal diagnosis of hemolytic disease of newborn (HDN) resulting from ABO blood group incompatibility. 4 pairs of primers were designed according to ABO blood group gene DNA and mRNA sequences. 20 plasma DNA samples from healthy donors were extracted and amplified to explore the best conditions for plasma DNA extraction and PCR amplification. The O group plasma DNA was mixed with A group or B group plasmas by the ratios of 1:1, 2:1, 4:1, 8:1, 10:1, 20:1, 40:1, 100:1 to simulate the status of mixed ABO gene from pregnant maternal blood and to establish the mixed blood group ABO genotyping technology. The pregnant maternal blood samples with more than 30 weeks of gestation were selected for detecting the fetal ABO blood group genotype. The blood samples should be taken as possible as after birth for identification of ABO blood group and evaluation of sensitivity and accuracy of fetal ABO blood group genotyping technology through peripheral blood of pregnant women. The results indicated that the minimal amount of template DNA from single blood plasma for accuracy identification was at least about 0.625 ng, the DNA amount extracted from 500 microl of plasma could meet the requirement for PCR amplification. When the proportion of O group plasma DNA in mixed plasma DNA was <or=10, the non-O group gene could be accurately detected. Among 14 peripheral blood samples from O-group pregnant women, the non-O group gene was amplified in 9 samples; the non-O group gene was not amplified in 5 samples. The identification of peripheral blood ABO group for 5 newborns using serologic method showed that the A group 3 cases, B group 2 cases, O group 1 case, which consisted with genotyping results with consistent rate 100%. It is concluded that in middle and late pregnancy the fetal ABO group gene can be detected accurately by means of established fetal ABO group gene extraction and typing technology, that provides some guidances for the prenatal diagnosis and prevention of HDN.

Fabrication and application of three-dimensional autogenous cartilage framework in auricular reconstruction / 中华整形外科杂志

<p><b>OBJECTIVE</b>To explore the fabrication and application of three-dimensional autogenous cartilage framework in auricular reconstruction.</p><p><b>METHODS</b>The process of fabrication of three-dimensional cartilage framework consisted of cartilage harvesting, carving and assembling the cartilage. The rib cartilage was harvested separately. The three-dimensional framework was composed of three main parts:the helix, the base and the pad, at different layer. The framework was fabricated according to the development of rib cartilage and contour and height of the reconstructed ear.</p><p><b>RESULTS</b>From January 1992 to December 2006, 5,248 patients of microtia were treated with 6,252 autogenous cartilage frameworks.</p><p><b>CONCLUSIONS</b>The three-dimensional framework is easily manipulated. The reconstructed ears look natural and had an erect contour. This method can effectively use the cartilage.</p>

Application of quantitative tissue expansion in ear reconstruction / 中华整形外科杂志

<p><b>OBJECTIVE</b>To explore the application of tissue expander in ear reconstruction and to deal with the complications.</p><p><b>METHODS</b>50 ml kidney-shape tissue expanders were implanted subcutaneously in the mastoid area. The drainage tube was removed 3 days after operation. The suture was removed 10 days later. Since 7 days after operation, 5 ml NS was injected into the expander every time, three times a week. The total injection volume was about 60 ml. After that, the expander was maintained for one month.</p><p><b>RESULTS</b>From January 1992 to December 2006, 5,248 patients of microtia were treated with 6,252 expander. After the maintaining period, the expanded skin was thin and well-vascularized. The complication rate was 7.79%, including hematoma, malunion and infection.</p><p><b>CONCLUSIONS</b>The quantitative tissue expansion is easily manipulated with few complications. It can provide hairless, thin skin with reliable blood supply for ear reconstruction.</p>

Study of methylation of promoter of EYA1 gene in microtia / 中华整形外科杂志

<p><b>OBJECTIVE</b>To explore the methylation of CpG islands in promoter of eye absent gene 1 (EYA1) in microtia.</p><p><b>METHODS</b>The methylation of CpG islands in EYA1 gene in 64 microtias and 36 healthy controls were measured using the technique of matrix-assisted laser desorption/ionization-time of flight.</p><p><b>RESULTS</b>The methylation of CpG_Unit3 and CpG_Unit5 of EYA1 gene in microtia were 0.09258 +/- 0.033846 and 0.0922 +/- 0.02379, respectively, which were significantly lower than those in control.</p><p><b>CONCLUSIONS</b>Hypomethylation in microtia may be related to the pathogenesis of the disease.</p>

Ear reconstruction using soft tissue expander in the treatment of congenital microtia / 中华整形外科杂志

<p><b>OBJECTIVE</b>To explore surgical procedure of congenital microtia.</p><p><b>METHODS</b>Ear reconstruction was made using soft tissue skin expander and autogenous rib cartilage framework.</p><p><b>RESULTS</b>Long time follow-up showed that the flap of reconstructed ear was ruddy, soft, with normal sensory function; cartilage framework had no degeneration, absorption and deformation. In addition, the reconstructed ears were coincidence with the normal side on location, form and dimension.</p><p><b>CONCLUSIONS</b>It was a good method for congenital microtia to use soft tissue skin expander together with autogenous rib cartilage framework at present time.</p>

External ear reconstruction using expanded postauricular scar flap and Medpor framework / 中华整形外科杂志

<p><b>OBJECTIVE</b>To investigate the possibility of external ear reconstruction using expanded postauricular scar flap and Medpor framework in burn cases.</p><p><b>METHODS</b>External ear reconstruction using expanded postauricular scar flap in combination with Medpor framework was performed in 17 cases whose ear had burn injury.</p><p><b>RESULTS</b>Of the 17 cases, 15 cases achieved success; 2 cases experienced partial exposure of the framework due to inadequate wrapping of the subcutaneous fascia flap and later injury. The longest follow-up was three years, and the final result was satisfactory.</p><p><b>CONCLUSIONS</b>External ear reconstruction using expanded postauricular scar flap in combination with Medpor framework is a reliable method for adult (over 25 years) who has ear defect from burn injury.</p>