OBJECTIVE:To establish a method for the content determination of verbascoside in Naosaitong honeyed pill. METHODS:HPLC was performed on the column of Zorbax ODX with mobile phase of acetonitrile-0.1% phosphoric acid(16∶84, V/V)at a flow rate of 1.0 ml/min,the detection wavelength was 334 nm,column temperature was 30 ℃,and the injection volume was 10 μl. RESULTS:The linear range of verbascoside was 0.200-1.000 μg(r=0.999 8);RSDs of precision,stability and repro-ducibility tests were lower than 1%;recovery was 95.16%-99.78%(RSD=1.80,n=6). CONCLUSIONS:The method is simple and accurate,and suitable for the content determination of verbascoside in Naosaitong honeyed pill.

Objective To investigate the expression of CD133 mRNA in tissues of gastric cancer,and explore its relationship with clinicopathological features. Methods The tissues of gastric cancer and normal tissues adjacent to gastric cancer were obtained from 31 patients.The expression of CD133 mRNA was detected by semi-quantitative RT-PCR,and its relationship with clinicopathological features such as sex,age,tumor diameter,infiltration depth,TNM staging,tumor differentiation,lymph node metastasis and Ki-67 proliferation index was analysed. Results The relative gray scale values of CD133 mRNA in tissues of gastric cancer and normal tissues adjacent to gastric cancer were 0.378 3±0.141 1 and 0.038 1 ±0.091 9,respectively(P=0.000).The relative gray scale values of CD133 mRNA in tissues of gastric cancer with tumor diameter>5 cm were significantly higher than those with tumor diameter≤5 cm[(0.439 3±0.148 4)vs(0.334 3±0.121 2)](P=0.041),and those in tissues with lymph node metastasis were significantly higher than those without lymph node metastasis[(0.426 6±0.132 0)vs(0.239 5±0.030 9)](P=0.004).The rate of lymph node metastasis and the number of metastatic lymph nodes were positively related to relative gray scale values of CD133 mRNA(r=0.466,P=0.008;r=0.464,P=0.009).The relative gray scale values of CD133 mRNA in those with low expression of Ki-67 were significantly higher than those with high expression of Ki-67[(0.436 4±0.139 8)vs(0.316 4±0.117 4)](P=0.02),and expression of Ki-67 were negatively related to relative gray scale values of CD133 mRNA(r=-0.461,P=0.009).Conclusion The expression of CD133 mRNA in tissues of gastric cancer was associated with the rate of lymph node metastasis,number of metastatic lymph nodes and expression of Ki-67,which reflect the status of lymph node metastasis and proliferation of gastric cancer.

OBJECTIVETo determine the physical and magnetic properties of superparamagnetic iron oxide (SPIO) nanoparticle prepared in our laboratory and evaluate its possibility for use as contrast agents in magnetic resonance imaging (MRI).

METHODSThe SPIO nanoparticle was obtained by means of classical coprecipitation in dextran solution and its size determined by electron microscopy and photon-correlation spectroscopy. The iron content was determined by phenanthroline photometry, and T(2) values as well as relaxivity evaluated with a clinical MR system at 1.5T.

RESULTSDextran-coated magnetite particles with a hydrodynamic diameter of 85.9 nm were prepared. The iron core size was 15 nm and the formation of Fe(3)O(4) crystal in SPIO nanoparticles was confirmed by X-ray diffraction (XRD) analysis. These particles possessed some characteristics of superparamagnetic and show a smaller spin-spin relaxation, with relaxivity and saturation magnetization of 0.1567 mmol(-1)/ms(-1) and 80 emu/g Fe, respectively.

CONCLUSIONSA stable SPIO nanoparticle with a dextran coating have been developed, and in vitro evaluation of its physical and magnetic properties suggests its potential for use as the contrast agent in MRI.

Humans ; Iron ; chemistry ; Magnetic Resonance Imaging ; methods ; Nanoparticles ; chemistry ; Oxides ; chemical synthesis ; chemistry ; X-Ray Diffraction

Objective:To identify and analyze different proteins expression in the periosteum of congenital pseudarthrosis of the tibia (CPT) using tandem mass tags (TMT) proteomics.Methods:The samples were divided into three groups, namely CPT with neurofibromatosis type 1 (NF1) group (NF1-CPT group), CPT without NF1 group (nonNF1-CPT group) and control group (patients with open tibial fracture). A fold change ≥1.5 or ≤0.66 and P-value <0.05 was regarded as the threshold to screen differentially expressed proteins (DEPs). Subsequently, bioinformatics resources such as online tools DAVID and STRING were used to conduct GO annotation, KEGG pathways enrichment and protein-protein interaction (PPI) network with DEPs. Results:A total of 347 proteins differentially expressed in NF1-CPT group, 212 of which were up-regulated and 135 down-regulated. We identified 467 DEPs in nonNF1-CPT group, including 281 up-regulated and 186 down-regulated. Among of them, NF1-CPT group and nonNF1-CPT group shared 231 DEPs, except for HLA-DRB1 which increased in NF1-CPT group but decreased in nonNF1-CPT group. The remaining 230 DEPs showed the same expression trend in the two positive groups, including 117 up-regulated and 113 down-regulated. In particular, a total of 116 proteins were altered only in NF1-CPT group, including 94 up-regulated and 22 down-regulated. However, there were 236 proteins altered only in nonNF1-CPT group, including 164 up-regulated and 72 down-regulated. The results indicated that the pathogenesis of NF1-CPT was similar as nonNF1-CPT largely with a few differences. Finally, compared with nonNF1-CPT, there were 47 proteins changed 1.5-fold and P-value <0.05 in NF1-CPT group. Conclusion:The proteins expression in the periosteum of CPT is different from that of normal tibia. The expression of periosteal protein is also different between NF1-CPT and nonNF1-CPT. The present study will deepen our understanding of the pathogenesis of CPT in the protein level.

Objective:To evaluate the effect of age on mood of patients during induction of general anesthesia with remimazolam.Methods:Eighty American Society of Anesthesiologists physical status Ⅰ or Ⅱpatients of both sexes, undergoing elective surgery with general anesthesia requiring tracheal intubation, aged 5-80 yr, with body mass index 18-30 kg/m 2, were divided into 4 groups ( n=20 each) according to different ages: juvenile group (R 1 group, 5-17 yr), youth group (R 2 group, 18-44 yr), middle-aged group (R 3 group, 45-59 yr) and elderly group (R 4 group, 60-80 yr). Remimazolam was intravenously injected in a loading dose of 0.1 mg/kg for induction of general anesthesia, followed by an infusion of 0.1 mg·kg -1·h -1 for maintenance of general anesthesia, and the other anesthetics for induction and maintenance of anesthesia were all the same in each group.The changes in emotion and brain wave within 3 min after remimazolam injection were recorded.The brain waves and Modified Observer′s Assessment of Alertness and Sedation (MOAA/S) scale score were recorded at 1, 2 and 3 min after administration.Adverse effects during anesthesia induction were also recorded. Results:Compared with group R 1, the incidence of happy mood was significantly decreased, the grade of mood was decreased, and the dream Likert score and MOAA/S score were decreased in R 3 and R 4 groups, and the dream Likert score and MOAA/S score were decreased in group R 2, and the incidence of hypoxemia during induction of general anesthesia was increased in group R 4( P<0.01). Compared with group R 2, the incidence of happy mood was significantly decreased in R 3 and R 4 groups, and MOAA/S score was significantly decreased in group R 4 ( P<0.05). Compared with R 1 and R 2 groups, the proportion of beta bands was significantly decreased, and the proportion of theta and delta bands was increased in R 3 and R 4 groups ( P<0.01). Compared with group R 3, the proportion of beta band was significantly decreased, and the proportion of delta band was increased in group R 4 ( P<0.01). Conclusion:Remimazolam 0.1 mg/kg used for induction of general anesthesia can cause happy mood, accompanied by enhanced brain electrical signal activity of beta band, especially in the patients under 45 yr of age.

Objective:To explore the anatomical and clinical effects of lateral-perineal approach in treating the fracture of inferior ramus of pubis-ischium ramus.Methods:The lateral approach of the perineum was simulated on 10 side of 5 intact wet adult cadavers to determine the surface symbols of incision design and to expose the operative field of the approach and to observe the anatomical characteristics of the perineal branch of the posterior femoral cutaneous nerve and the incision of the surgical approach. Five points were selected at the incision of the approach. The distance (L 1-L 5) between each point and the body surface projection of the perineal branch of the posterior femoral cutaneous nerve was measured. A total of 11 patients, including 5 males and 6 females with an average age of 41.55±14.32 years, ranging from 18 to 62 years, were treated by this approach in clinical practice. All patients had a reduction and fixation to the fracture of inferior ramus of pubis-ischium ramus. The operation duration, incision length, intraoperative blood loss and surgical complications were recorded. The quality of fracture reduction was evaluated according to Matta radiographic criteria. The strength of the adductor was measured. The pelvic function was evaluated according to Majeed Pelvic Score at the last follow-up. Results:Anatomical studies shown that the line between the two points. One point was 4 cm lateral to the level of the apex of the pubic arch. Another point was 4 cm from the ischial tubercle on the line from the ischial tuberosity to the point that 4 cm lateral to the level of the apex of the pubic arch was the axis of the approach lateral of the perineum. Anatomical studies showed that the lateral-perineal approach could expose the range from pubic symphysis to sciatic tuberculum. The distance between the points selected at the incision of the approach and the body surface projection of the perineal branch of the posterior femoral cutaneous nerve: L 1 was 19.40±1.17 mm, ranging from 18 to 21 mm; L 2 was 16.60±2.76 mm, ranging from 10 to 20 mm; L 3 was 18.30±1.89 mm, ranging from 16 to 21 mm; L 4 was 19.20±1.93 mm, ranging from 16 to 22 mm; L 5 was 14.70±1.83 mm, ranging from 13 to 18 mm. All patients were followed up for 17.91±4.09 months, ranging from 13 to 26 months. The incision length was 8.18±0.98 cm, ranging from 7 to 10 cm. The operation duration was 59.64±12.17 min, ranging from 43 to 85 min. The intraoperative blood loss was 100 ml, ranging from 50 to 130 ml. All incisions were healed in all patients. The fractures were healed in 13.36±2.06 weeks, ranging from 10 to 16 weeks. According to Matta radiographic criteria, the quality of fracture reduction was excellent in 6 cases, good in 4 cases and fair in 1 case. At the last follow-up, the adductor muscle strength reached grade 4 in 4 patients and grade 5 in 7 patients. Furthermore, according to the Majeed Pelvic Score, the score of every patient was 86.55±9.59, ranging from 66 to 100, and 8 cases were excellent, 3 cases were good at the last follow-up. The heterotopic ossification occurred in 2 patients, the pain during intercourse occurred in 2 patients. No patient had sensory disturbance or pain in the perineal area. Conclusion:A certain safe distance is between the lateral to the perineum and the perineal branch of the posterior femoral cutaneous nerve with limited risk of injuring posterior femoral cutaneous nerve via the lateral approach of the perineum. The advantages in treating the fracture of inferior ramus of pubis-ischium ramus by this approach have concealed incision, short operation duration and less bleeding with satisfied short-term clinical effects.

Objective:To develop a model of stellate ganglion block (SGB) in mice and investigate the effect of SGB on cerebral cortical blood flow.Methods:Thirty clean-grade healthy male C57BL/6 mice, aged 8-9 weeks, weighing 23-27 g, were divided into 5 groups ( n=6 each) using a random number table method: control group (group C), left SGB group (group L), left normal saline group (group SL), right SGB group (group R) and right normal saline group (group SR). Group C received no intervention.SGB was performed with 0.25% ropivacaine 0.08 ml via percutaneous posterior approach in L and R groups, while the equal volume of normal saline 0.08 ml was given instead at the location of left and right stellate ganglion in SL and SR groups, respectively.The cerebral cortical blood flow was determined using laser speckle contrast imaging system before SGB (T 0) and at 10, 30, 60, 90 and 120 min after SGB (T 1-5). Results:Mice developed ptosis on the block side, indicating that the model of SGB was successfully developed in L and R groups.There was no significant difference in cerebral cortical blood flow at each time point among C, SL and SR groups ( P>0.05), and cerebral cortical blood flow on the block side decreased at T 1, began to increase at T 2, peaked at T 3, and decreased at T 5 which was still higher than that at T 0 in group L and group R ( P<0.01). Compared with C and SL groups, the left cerebral cortical blood flow was significantly decreased at T 1, 5 and increased at T 2-4 in group L ( P<0.01). Compared with C and SR groups, the right cerebral cortical blood flow was significantly decreased at T 1, 5 and increased at T 2-4 in group R ( P<0.01). There were no significant differences in cerebral cortical blood flow at each time point between group C and group SL and between group C and group RL ( P>0.05). Conclusions:The mouse model of SGB via percutaneous posterior approach is successfully developed.Unilateral SGB can affect cerebral cortical blood flow on the block side, which shows a transitory decrease followed by a sustained significant increase.

Pancreatic neuroendocrine neoplasms (pNENs) are highly heterogeneous, and the management of pNENs patients can be intractable. To address this challenge, an expert committee was established on behalf of the Chinese Pancreatic Surgery Association, Chinese Society of Surgery, Chinese Medical Association, which consisted of surgical oncologists, gastroenterologists, medical oncologists, endocrinologists, radiologists, pathologists, and nuclear medicine specialists. By reviewing the important issues regarding the diagnosis and treatment of pNENs, the committee concluded evidence-based statements and recommendations in this article, in order to further improve the management of pNENs patients in China.

OBJECTIVE：To study the potential mechanism of yam protein （DOT） in the prevention and treatment of diabetes-induced erectile dysfunction （DIED）. METHODS ：DIED model was induced by high-glucose and high-fat diet and intraperitoneal injection of streptozotocin （40 mg/kg）. The experiment was set up in the normal control group （normal saline ）， model group （normal saline ），DOT low-dose ，medium-dose and high-dose groups （0.3，0.6，0.9 mg/kg），sildenafil group （positive control ，4.4 mg/kg），with 9 rats in each group. In the stage of successful establishment of diabetes model and initiation of inducing DIED ，rats in each group were given relevant solution intragastrically ，once a day ，for consecutive 11 weeks. Body weight，fasting plasma glucose （FPG），the times and rate of penile erection ，fasting insulin （FINS），insulin resistance index （IR），the contents of endothelial nitric oxide synthase （eNOS）and cyclic guanosine monophosphate （cGMP）in penile cavernous tissue were determined so as to evaluate the intervention effects of DOT on DIED model rats. High-glucose damaged mice cavernous endothelial cells （MCECs）model was induced by 30 mmol/L glucose for 48 h，and then give DOT 125，250，500 μg/mL. The cell viability was detected so as to evaluate the effects of DOT on high-glucose damaged MCECs model. RNA-Seq mail：xingxin0902@163.com technology was adopted to screen the differentially expressed genes between normal MCECs and high-glucose damaged MCECs，high-glucose damaged MCECs and MCECs treated with 250 μg/mL DOT. Gene ontology（GO）function enrichment analysis and KEGG pathway enrichme nt analysis were performed for differentially expressed genes. The common differentially expressed genes between 2 groups were analyzed ，and mRNA expressions of six key genes were validated. RESULTS ：Different doses of DOT could reverse the reduction of body weight ，the increase of FINS and IR ，the reduction of the times and rate of penile erection ，the decrease of eNOS and cGMP contents in penile cavernous tissue of DIED model rats ；above indexes of DIED model rats were reversed significantly after treated with high-dose of DOT（P＜0.05 or P＜0.01）. 125，250，500 μg/L DOT could significantly improve the activity of high-glucose damaged MCECs （P＜0.05 or P＜0.01）. RNA-Seq technology showed that compared with normal MCECs ，a total of 48 differentially expressed genes were found in high-glucose damaged MCECs. Compared with high-glucose damaged MCECs ，a total of 779 differentially expressed genes were found in MCECs treated with DOT. The differentially expressed genes of 2 groups were mainly cellular process in biological process annotation ，cellular part in cell component annotation and binding molecular function in molecular function annotation ，which were mainly enriched in extracellular matrix receptor interaction pathway ，mismatch repair pathway ， phosphatidylinositol 3 kinase-protein kinase B （PI3K-Akt）signal pathway and so on. Among differentially expressed genes of 2 groups，13 common differentially expressed genes such as Aldh1a1，Abcc5，Tac1 were found. DOT could significantly reverse the expression of the above common differentially expressed genes in high-glucose damaged MCECs. After validation ，DOT could significantly reverse the mRNA expression of TGF-β3，Txnip，Aldh1a1，Loxl1，Mt1 and Mt2 in high-glucose damaged MCECs. CONCLUSIONS：DOT could improve the symptom of DIED model rats ，the mechanism of which may be related to biological pathway of inhibiting fibrosis and reducing oxidative stress ，so as to improve the endothelial function of cavernous body.

Objective:To investigate the genetic safety of allogeneic bone marrow mesenchymal stem cells (BMSCs) transplantation in traumatic brain injury (TBI).Methods:(1) In vivo experiment: BMSCs from male SD rats were isolated and cultured. Moderate TBI models were prepared by implanting and fixing micro-drug injection cannula into the left ventricle of 12 female SD rats, and 3 d after that, striking the right cerebral cortex of the rats with pneumatic precision percussion device was performed. Four h, and 3, 6, 9, and 12 d after modeling, TBI rats were given a single/multiple BMSCs infusion (2.5×10 5/time, total volume 10 μL) by cannula; 48 and 72 h, and 10 and 14 d after modeling, brain tissues of TBI rats (3 at each time point) were prepared into paraffin specimens. Immunofluorescent staining was used to detect the microglia activation, and RNAscope ? technology was used to detect the co-localization of astrocytes, neurons, microglia and transplanted BMSCs to observe whether the allogeneic BMSCs were integrated with the host brain cells after transplantation into TBI host. (2) In vitro experiment: the frozen and revived microglial cell line BV2 was transfected with green fluorescent protein (GFP)-positive lentiviral particles, and then, BMSCs prelabeled with pHrodo RED probe and BV2 cells pretreated with lipopolysaccharide were co-cultured in a certain ratio (BV2:BMSCs=1:1, 1:2, 2:1); after 36 and 72 h of co-culture, the phagocytosis between the 2 kinds of cells was observed under confocal fluorescence inverted microscope to observe the specific action forms of microglia on BMSCs. Results:(1) In vivo experiment: 48 and 72 h, and 10 and 14 d after modeling, no colocalization of transplanted BMSCs with astrocytes or neurons was found in paraffin sections of brain tissue in TBI rats; however, 10 and 14 d after modeling, microglia in TBI rats were obviously activated and migrated to the left lateral ventricle and choroid plexus, and co-localization of microglia with transplanted BMSCs was observed. (2) In vitro experiment: phagocytosis occurred after co-culture of BV2 cells at different proportions with BMSCs for 36 and 72 h. Conclusion:After transplantation, allogeneic BMSCs do not integrate with astrocytes or neurons of the TBI host, but they could be phagocytosed by microglia, indicating that allogeneic BMSCs transplantation for TBI is genetically safe.