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Objective:To observe the effect of microRNA-217 (miR-217) on neuropathic pain and neuro-inflammation in rats with partial sciatic nerve ligation (pSNL).Methods:Twenty-four healthy adult male SD rats were randomly divided into three groups: pSNL group (partial sciatic nerve ligation), Sham group (Sham surgery), and normal group (no treatment) with 8 in each group. The mechanical withdrawal threshold (MWT) and paw withdrawal thermal latency (PWTL) were measured on the 1st, 2nd and 3rd day before surgery and on the 1st, 3rd, 7th and 14th day after surgery.The mRNA expression changes of miR-217 and TLR5 in the spinal cord of rats were detected by qRT-PCR. Additionally, 24 healthy adult male SD rats were randomly divided into three groups: pSNL- LV-miR-217 group (sciatic nerve part ligation + intrathecal injection of LV-miR-217), pSNL- LV-NC group (sciatic nerve part ligation + intrathecal injection of LV-NC), and Sham operation group with 8 in each group. MWT and PWTL were measured on the first, second and third day before surgery and on the 1st, 3rd, 7th, 14th day after surgery.Results:The results of pain behavior showed that the MWT of SNL group decreased significantly on the 7th day ((6.72±0.15) g, t=11.79, P<0.05) and the 14th day ((5.72±0.22) g, t=17.83, P<0.05), compared with sham group (7th d (13.58±0.30) g, 14th d (14.54±0.51)g). PWTL of pSNL group was (6.78±0.21) s and (5.99±0.22)s respectively on the 7th and 14th day after operation, which was lower than those of Sham group ((14.86±0.35) s, (14.50±0.51) s), the difference was statistically significant ( t=4.86, 7.13, both P<0.05). The qPCR experiment showed that the expression of miR-217 was down-regulated and the expression of TLR5 was up-regulated in the spinal cord of rats in the pSNL group, showing significant differences compared with the Sham group ( t=8.94, P<0.01; t= 7.54, P<0.01). The MWT of the pSNL- LV-miR-217 group were significantly increased on the 7th and 14th day after the intraoperative injection of miR-217, showing a significant difference compared with the pSNL-LV-NC group (7th d: t=14.79, P<0.01; 14th d: t=15.83, P<0.01). The PWTL of the pSNL-LV-miR-217 group were significantly increased(7th d: t=14.95, P<0.01; 14th d: t=16.27, P<0.01). qPCR assay showed that compared with the pSNL-LV-NC group, the expression of miR-217 in the pSNL-LV-miR-217 group was significantly increased ( t=7.49, P<0.01), and the expression of TLR5, COX-2( t=8.82, P<0.05), IL-6( t=6.42, P<0.05) and TNF-α( t=7.46, P<0.05) was significantly decreased. Western blot assay showed that the protein expressions of TLR5, COX-2, IL-6 and TNF-α in pSNL-LV-miR-217 group were significantly decreased on the 7th day after the operation. Conclusions:Intrathecal injection of miR-217 can significantly alleviate sciatic nerve ligation neuropathic pain and may inhibit neuroinflammation by regulating TLR5.
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Ophiocordyceps sinensis is a valuable traditional Chinese medicine. Due to resource shortage, expensive price and huge market demand, there are many adulterants of O. sinensis in markets. Therefore, it is necessary to establish a rapid and effective method for distinguishing O. sinensis. Based on the species-specific PCR of O. sinensis, this study developed a detection kit by optimizing the components and evaluated the specificity, detection limit, repeatability and shelf life of the kit. The results showed that when the quality of O. sinensis accounted for more than 1/200 of that mixture, it could be detected successfully. Moreover, only O. sinensis could be amplified and glowed bright green fluorescence under ultraviolet light. The kit was still in effect when it was placed at 37 ℃ for three days, which indicated that it was stable and effective for one year stored in 4 ℃. The kit in the same batch under different operation conditions, and in different batch under the same operation conditions gave the same result and accuracy, which showed good repeatability of the kit. It is simple, rapid and accurate to distinguish O. sinensis from its adulterants using the kit, and lays the foundation for commercialization of traditional Chinese medicine fast detection kit.
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AIM:To detect the expression of miRNA-363 and SOX4 in osteosarcoma tissues and to investigate the effect of miRNA-363 on the viability and apoptosis of human osteosarcoma cell line MG-63.METHODS: Real-time PCR was used to detect the expression level and the relationship of miRNA-363 and SOX4 mRNA in the osteosarcoma tis-sues and the corresponding paratumorous tissues collected from 63 patients.The expression levels of miRNA-363 and SOX4 in osteosarcoma cell line MG-63 after transfected with miRNA-363 mimics were measured .The cell viability was measured by CCK-8 assay.Flow cytometry was used to monitor the changes of cell cycle and apoptosis .The changes of SOX4 and miRNA-363 expression levels in the MG-63 cells after transfection with SOX4 siRNA or pcDNA/SOX4 was detect by real-time PCR.RESULTS:The expressed level of miRNA-363 was lower , and the expression level of SOX 4 was higher in the osteosarcoma tissues than those in the adjacent normal tissues .A significantly negative correlation between the expression levels of miRNA-363 and SOX4 was observed .The expression of miRNA-363 in the MG-63 cells after transfection with miRNA-363 mimics was significantly up-regulated, while the expression of SOX4 in the MG-63 cells was significantly down-regulated , with significant difference as compared with the cells transfected with miRNA-NC and control cells .The viability of MG-63 cells was inhibited, the cell cycle was arrested in G0/G1 phase, and the cell apoptosis was increased by transfec-tion with miRNA-363 mimics.The relative protein expression levels of SOX 4 in SOX4 siRNA group and pcNDA/SOX4 group were significantly different from those in negative control group , but the relative expression levels of miRNA-363 had no significant difference .Over-expression of SOX4 restored the viability of the MG-63 cells reduced by miR-363.CON-CLUSION:The expression level of miRNA-363 is low in human osteosarcoma tissue .miRNA-363 may inhibits the viabili-ty of osteosarcoma cell line MG-63 and promotes cell apoptosis in vitro via inhibiting the SOX4 expression.
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Objective To establish a small intestinal organoid culture system as an in vitro study model of intesti-nal epithelial cells, and to explore the relevant pathological detection techniques and provide a convenient platform for in vitro study of various intestinal diseases. Methods The mouse intestinal epithelium was isolated and cultured into or-ganoids to simulate the growth and development of intestinal epithelium in vitro. The proliferation and differentiation signals were detected by immunohistochemistry and three-dimensional immunofluorescence technique. Results The culture system of the mouse small intestine epithelium was established. Immunohistochemical staining and three-dimensional immunofluo-rescence technique were successfully used to detect the growth and development of small intestinal organoids. Conclusions The successfully established mouse small intestinal organoid culture system and application of immunoassay technology will gradually become a most favorable technical means for studies of various intestinal diseases.
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Objective To establish a small intestinal organoid culture system as an in vitro study model of intesti-nal epithelial cells, and to explore the relevant pathological detection techniques and provide a convenient platform for in vitro study of various intestinal diseases. Methods The mouse intestinal epithelium was isolated and cultured into or-ganoids to simulate the growth and development of intestinal epithelium in vitro. The proliferation and differentiation signals were detected by immunohistochemistry and three-dimensional immunofluorescence technique. Results The culture system of the mouse small intestine epithelium was established. Immunohistochemical staining and three-dimensional immunofluo-rescence technique were successfully used to detect the growth and development of small intestinal organoids. Conclusions The successfully established mouse small intestinal organoid culture system and application of immunoassay technology will gradually become a most favorable technical means for studies of various intestinal diseases.
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In order to investigate the content and distribution of available element in the rhizonsphere soil of the growing areas of Salvia miltiorrhiza Bunge, the contents of available element (N,P,K,B,Cu,Zn,Fe,Mn) in 26 soil samples were tested and evaluated. The results showed that the contents of available P and Fe were very plentiful, available K, Cu and Zn were rich, available N and Mn were deficient, available B was extremely deficient in all growing areas of S. miltiorrhiza of eight provinces in China. The correlation analysis showed that the contents of eight kinds of available elements were varying degree correlation. The stepwise regression analysis between the contents of available elements of rhizonsphere soil and ten kinds of active ingredients of Danshen (Salviae Miltiorrhizae Radix et Rhizoma) were researched. The results showed that the rates of contribution of available N,B,Mn and Fe to quality of Danshen were relatively large and they were the significant factors, and the other factors did not show statistical significance. The recommended fertilizing strategies is that the usage of N,B and Mn fertilizers should be controlled according to different stages of growth of S. miltiorrhiza, and P fertilizer should be reduced in all growing areas of S. miltiorrhiza.
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To study the compatible mechanisms and compatible proportion of Shaoyao Gancao decoction, the intestinal absorption of main ingredients in Shaoyao Gancao decoction SG11 (Baishao-Zhigancao 1: 1) , SG31 (Baishao-Zhigancao 3: 1), Baishao water decoction S and Zhigancao (G) were investigated and compared using in vitro everted intestinal sac model and in situ single pass intestinal perfusion (SPIP) model. The concentration of paeoniflorin (PF), liquiritin (LQ) and mono-ammonium glycyrrhizinate (GL) in test samples and samples of intestinal sac and intestinal perfusion was determined by HPLC. The intestinal absorptive amount and absorption parameters were calculated. Results showed that in the everted intestinal sac model, three ingredients could be absorbed by duodenum, jejunum and ileum, and the absorption in the jejunum was best for all 3 ingredients. The absorption rate of three ingredients in SG11 was significantly higher than that in single decoction (P < 0.05), but had no significant difference compared with SG31. In SPIP model, the absorption rate constant K(a), the apparent absorption coefficient P(app) and the absorption rate of three ingredients in SG11 were significantly higher than those in single decoction. Parameters of PF and GL in SG11 were significantly higher than those in SG31, but had no differences of LQ. It proved that the compatibility of Baishao and Zhigancao could improve the intestinal absorption of PF, LQ and GL. The absorption of each ingredient in SG11 was better than that in SG31.
Subject(s)
Animals , Male , Rats , Chromatography, High Pressure Liquid , Drugs, Chinese Herbal , Pharmacokinetics , Intestinal Absorption , Intestines , Metabolism , Rats, Sprague-DawleyABSTRACT
BACKGROUND:The treatment of limbs’ open fracture with severe soft tissue injury is very complicated. There are many wound exudates, high frequency of dressing change, and high infection rate. How to deal with the contradiction between wound treatment and fracture fixation is the considerable problem in clinical therapy. ＜br> OBJECTIVE:To explore the effect of external fixator combined with vacuum sealing drainage on limbs’ open fracture with severe soft tissue injuries. ＜br> METHODS:A total of 34 patients with limbs’ open fracture with severe soft tissue injuries were enrol ed in the First Affiliated Hospital, Xinxiang Medical Col ege from December 2009 to December 2010. There were 20 males and 14 females, with an average age of 36.6 years old. There were 10 cases of upper limb fracture and 24 cases of lower limb fracture. After debridement, cutaneous deficiency accounted for 1%to 6%of surface area. On the basis of external fixation, the wound received vacuum sealing drainage (treatment group;n=17) and conventional dressing (control group;n=17). According to limb swel ing and granulation growth, delayed suture, skin graft or flap to cover the wound were used. After treatment, infection rate of the wound and growth of granulation were observed. In accordance with wound healing and growth of granulation, curative effects were evaluated. ＜br> RESULTS AND CONCLUSION:In the treatment group, there were 11 cases of healing, 5 cases of effective effects, and 1 case of ineffective effect, with a total effective rate of 94%and the infection rate of about 15%. In the control group, there were 7 cases of healing, 6 cases of effective effects, and 4 cases of ineffective effects, with a total effective rate of 76%and the infection rate of about 40%. Significant differences in healing rate, total effective rate and infection rate were detected between the two groups (P＜0.05). The healing time of wound was 8.66-16.23 days in the treatment group and 15.68-22.36 days in the control group. The healing time was significantly shorter in the treatment group than in the control group (P＜0.05). It is thus clear that vacuum sealing drainage has obvious advantages in repair of limbs’ open fracture with severe soft tissue injury compared with the conventional dressing.
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<p><b>OBJECTIVE</b>To establish an HPLC fingerprint to evaluate the quality of Polygalae Radix, root xylem, and those collected in different growth ages or harvest time.</p><p><b>METHOD</b>Separation was performed at 30 °C on a Kromasil C18 column (4.6 mm x 250 mm, 5 μm); the mobile phases was acetonitrile and 0.05% H3PO4 water in the gradient elution; the flow rate was set at 1.0 mL · min(-1) and the detection wavelength at 314 nm; the quality discriminant analyses were accomplished by means of similarity analysis, cluster analysis, principal component analysis and neural network model.</p><p><b>RESULT</b>In 26 batches of Polygalae Radix, 24 batches fingerprint similarities were above 0.8. In 5 different growth or harvest time batches, 4 batches were above 0.8; in 8 batches root xylem samples, the similarities were all above 0.875. The similarity analysis was in accord with the quality discriminant analysis of cluster analysis, principal component analysis and neural network model.</p><p><b>CONCLUSION</b>Fingerprint combined with chemical pattern recognition technique can effectively evaluate the quality of Polygalae Radix. The active substance species are all similar in cultivated, wild, different growth or harvest time Polygalae Radix and polygala root xylem, but the chromatography peak areas are different. The effective material contents are similar between wild and cultivated Polygalae Radix, but each chromatographic peak area of the root xylem is much smaller than that of Polygalae Radix. The chemical substance accumulation mainly depends on harvest month, but little growth time in Polygalae Radix.</p>
Subject(s)
Chromatography, High Pressure Liquid , Methods , Drugs, Chinese Herbal , Chemistry , Plant Roots , Chemistry , Classification , Polygala , Chemistry , Classification , Quality ControlABSTRACT
Mantidis Oötheca is commonly used Chinese medicine. Because of the used medicinal part is oötheca and many mantis species can yield ootheca, it is not possible to identify its original animal accurately. There is no unanimous conclusion about the corresponding relationship between Mantis and Mantidis Oötheca (Sangpiaoxiao). This relationship is the basis of the Mantidis Oötheca research. Our study combined the methods of artificial incubation oötheca and capture the living mantis to identify the species of Mantis and Mantidis Oötheca. The results showed that the origin insects of Mantidis Oötheca was Tenodera, Hierodula and Statilia genus insects. This has laid a foundation for further study of Mantidis Oötheca.
Subject(s)
Animals , Mantodea , Chemistry , Classification , Medicine, Chinese Traditional , Reference Standards , Quality ControlABSTRACT
<p><b>OBJECTIVE</b>To clarify the commercial original species of hippocampus in the market.</p><p><b>METHOD</b>Field survey and interview were applied to the investigation.</p><p><b>RESULT</b>Present study identified the main commercial hippocampus of 13 species, including Pharmacopoeia contained four kinds of hippocampus (without Hippocampus japonicus) listed in Chinese Pharmacopoeia, and the identification method for the 13 kinds of commercially hippocampus was established.</p><p><b>CONCLUSION</b>The further research on hippocampus should be strengthened for the establishment of hippocampal quality control standard.</p>
Subject(s)
Animals , Medicine, Chinese Traditional , Economics , Reference Standards , Quality Control , Smegmamorpha , ClassificationABSTRACT
To analyze the content and type of soluble proteins in Cordyceps sinensis from different producing areas and processed with different methods with bradford method and 2-DE technology, in order to discover significant differences in soluble proteins in C. sinensis processed with different methods and from different producing areas. The preliminary study indicated that the content and diversity of soluble proteins were related to producing areas and processing methods to some extent.
Subject(s)
Cordyceps , Chemistry , Proteins , Chemistry , SolubilityABSTRACT
<p><b>OBJECTIVE</b>To understand the etiology of hand, foot and mouth disease (HFMD) in Guangzhou area in 2008.</p><p><b>METHOD</b>Totally 1023 clinical specimens were collected from pediatric patients suspected of HFMD in 2008. TaqMan real-time RT-PCR were used for detection of enterovirus 71 (EV71), Coxsackievirus A16 (CA16) and other enteroviruses. The specimens which were enterovirus positive by RT-PCR method with universal primer but EV71 and CA16 negative, were amplified and sequenced for 5'untranslated region.</p><p><b>RESULT</b>Enterovirus was identified from 434 of 1023 samples and detection rate of enterovirus was 42.42%; of the 434 samples, 276 were positive for EV71 (63.6%), 126 for CA16 (29%), 4 samples for enterovirus 84, 3 for Echovirus 11, 2 for Echovirus 9, 3 for Coxsackievirus B3, 4 for Coxsackievirus A10, 3 for Coxsackievirus A6, 6 for Coxsackievirus A12 or A5, and for 7 samples typing was difficult.</p><p><b>CONCLUSION</b>The major causative agents of HFMD in Guangzhou were EV71 and CA16 in 2008, and EV84, CA10, CA12, CA6, COSB3, ECHV11, ECHV9 were also the pathogens for smaller proportions of patients.</p>
Subject(s)
Child , Child, Preschool , Female , Humans , Infant , Male , China , Epidemiology , Coxsackievirus Infections , Epidemiology , DNA Primers , Enterovirus A, Human , Classification , Genetics , Hand, Foot and Mouth Disease , Epidemiology , Virology , RNA, Viral , Reverse Transcriptase Polymerase Chain ReactionABSTRACT
<p><b>OBJECTIVE</b>To analyze the change in nitration tyrosine, NO(2)(-)/NO(3)(-)level in induced sputum of silicosis patients and dust exposure workers and to evaluate the approach and feasibility of nitric oxide (NO) metabolites as early detection indicators of silicosis.</p><p><b>METHODS</b>Nitration tyrosine, NO(2)(-)/NO(3)(-)concentration in induced sputum of 80 dust exposure workers, 84 silicosis patients, 30 logistic personnel with no history of exposure to silica dust were determined and the relationship among Nitration tyrosine, NO(2)(-)/NO(3)(-)level and dust exposure years as well as pulmonary function tests were analyzed.</p><p><b>RESULTS</b>NO(2)(-)/NO(3)(-)level among exposed group [60.30 (46.58) micromol/l] was significantly higher than the control group [36.90 (22.28) micromol/l], (P < 0.05), and the level of NO(2)(-)/NO(3)(-)among the cases [79.65 (89.10) micromol/l] was significantly higher than exposed group as well as the control group (P < 0.05). Compared with control, the level of nitration tyrosine in exposed group [3.51 (0.46) nmol/l] and the cases [3.48 (0.49) nmol/l] was significantly higher (P < 0.05). NO(2)(-)/NO(3)(-)level and dust exposure years were positively correlated (r = 0.3733 and 0.3830 respectively P < 0.05); NO(2)(-)/NO(3)(-)level and pulmonary function tests (FVC%, FEV1.0%, PEF%, MEF25%, MEF50%) were negatively correlated (r = 0.1540, 0.1723, 0.1535, 0.1485, 0.1643 respectively, P < 0.05). There was no correlation between nitration tyrosine and dust exposure years (P > 0.05), no correlation between nitration tyrosine and pulmonary function test (P > 0.05).</p><p><b>CONCLUSION</b>The level of NO(2)(-)/NO(3)(-)level in induced sputum has a positive correlation with exposure to dust, suggesting that there will be a certain feasibility of the NO(2)(-)/NO(3)(-)as indicators of early detection of silicosis.</p>
Subject(s)
Adult , Humans , Middle Aged , Nitrates , Metabolism , Nitrites , Metabolism , Silicosis , Metabolism , Sputum , Metabolism , Tyrosine , MetabolismABSTRACT
A comprehensive analysis was conducted on resources composition of Chinese Herbs Sang (Morus sp.) in Sichuan using survey data and related literature. The original plants, germplasm collections, cultivation areas, main cultivated varieties and production sale of crude drugs of Sang in Sichuan were clearly expounded. Strategies for development and utilization of Sichuan mulberry resources were suggested.
Subject(s)
Biotechnology , China , Conservation of Natural Resources , Methods , Gardening , Methods , Morus , Plants, MedicinalABSTRACT
<p><b>OBJECTIVE</b>To investigate the correlation between entophytic fungal community, habitations and varieties of Ligusticum.</p><p><b>METHOD</b>The solidified potato dextrose agar (PDA) of plates was applied for the isolation of the endophytic fungi, and the identification was completed by spot-planting method.</p><p><b>RESULT</b>Fifty strains of the entophytic fungi were isolated from the rhizome of L. chuanxiong collected from 6 habitations. They were morphologically identified as belonging to 13 genera, 4 families, 3 orders and 1 class.</p><p><b>CONCLUSION</b>There were some differences at quantity, species and distributing of the entophytic fungi in different habitats and varieties of Ligusticum, which was suggested that entophytic fungal community is related with specific habitat.</p>
Subject(s)
Fungi , Classification , Ligusticum , Microbiology , Plants, Medicinal , MicrobiologyABSTRACT
<p><b>OBJECTIVE</b>To establish the HPLC fingerprint of Rhizoma Polygoni Cuspidati (Polygonum cuspidatum).</p><p><b>METHOD</b>The HPLC separation was carried with Diamonsil C18 column and eluted with a gradient from methanol and 0.1% phosphoric acid, the detection wavelength was at 230 nm and recording 70 min. The similarity of chromatograms was compared by mean of the software from Zhongnan University.</p><p><b>RESULT</b>The constituents of P. cuspidatum were well separated by HPLC, and the similarity was above 0.80.</p><p><b>CONCLUSION</b>The method can be used for the study of fingerprints of Rhizoma Polygoni Cuspidati.</p>
Subject(s)
Chromatography, High Pressure Liquid , Methods , Emodin , Fallopia japonica , Chemistry , Classification , Glucosides , Plants, Medicinal , Chemistry , Classification , Quality Control , Reproducibility of Results , Rhizome , Chemistry , Stilbenes , Technology, Pharmaceutical , MethodsABSTRACT
<p><b>OBJECTIVE</b>To study the phamacokinetics and relative bioavailability of combination Radix Angelicae Sinensis and Cortex Cinnamomi.</p><p><b>METHOD</b>The content of ferulic acid in plasma was determined directly by HPLC after oral administration of Cortex Cinnamomi in combination individually with Radix Angelicae Sinensis. The plasma concentration-time curve were plotted. The main pharmacokinetic parameters and relative bioavailability were obtained.</p><p><b>RESULT</b>The plasma concentration-time curve of fenalic aclid conformed to one-compartment model. The relative bioavailability of Cortex Cinnamomi combined with Radix Angelicae Sinensis were 226.75%.</p><p><b>CONCLUSION</b>By the statistical analysis, Cortex Cinnamomi combined with Radix Angelicae Sinensis can increase the relative bioavailability of ferulic acid.</p>