[Governor vessel moxibustion combined with warming acupuncture for knee osteoarthritis with yang deficiency and cold congelation based on the supporting-yang theory].

OBJECTIVE: To compare the clinical efficacy between governor vessel moxibustion combined with warming acupuncture based on the supporting-yang theory and simple warming acupuncture for knee osteoarthritis (KOA) with yang deficiency and cold congelation. METHODS: A total of 64 patients with KOA of yang deficiency and cold congelation were randomized into a combination group (32 cases, 2 cases dropped off) and a warming acupuncture group (32 cases, 1 case dropped off). In the warming acupuncture group, warming acupuncture was applied at Zusanli (ST 36), Guanyuan (CV 4) and Dubi (ST 35), Neixiyan (EX-LE 4), etc. on the affected side, once a day. On the basis of the treatment in the warming acupuncture group, governor vessel moxibustion was applied in the combination group, once a week. The 14-day treatment was taken as one course, and totally 2 courses with 2-day interval were required in the two groups. The clinical symptom score, the visual analogue scale (VAS) score and the Western Ontario and McMaster Universities arthritis index (WOMAC) score were observed before treatment, after treatment and in the follow-up of 12 weeks after treatment; the volume of suprapatellar bursa effusion was detected before and after treatment; the clinical efficacy was evaluated after treatment and in the follow-up in the two groups. RESULTS: After treatment and in the follow-up, the scores of clinical symptom, VAS and WOMAC were decreased compared before treatment in both groups (P<0.05), and those in the combination group were lower than the warming acupuncture group (P<0.05). After treatment, the volume of suprapatellar bursa effusion was decreased compared before treatment in both groups (P<0.05). After treatment and in the follow-up, the total effective rates were 93.3% (28/30) and 86.7% (26/30) in the combination group, which were superior to 87.1% (27/31) and 74.2% (23/31) in the warming acupuncture group respectively (P<0.05). CONCLUSION: Governor vessel moxibustion combined with warming acupuncture can improve the clinical symptoms i.e. pain and dysfunction and reduce the volume of suprapatellar bursa effusion in KOA patients with yang deficiency and cold congelation, its short-term effect and long-term effect are both superior to simple warming acupuncture.

Effects and mechanisms of foliar application of silicon and selenium composite sols on diminishing cadmium and lead translocation and affiliated physiological and biochemical responses in hybrid rice (Oryza sativa L.) exposed to cadmium and lead.

Currently, exploring effective measures to reduce multiple toxic metals accumulation in rice grains is an urgent issue to be tackled. Pot experiments were thus conducted to explore the effects and mechanisms of foliar spraying with composite sols of silicon (Si) and selenium (Se) during tillering to booting stage on diminishing cadmium (Cd) and lead (Pb) translocation to rice grains and affiliated physiological and biochemical responses in rice seedlings grown in Cd + Pb-polluted soils (positive control). Results showed that Cd and Pb contents in leaves or grains were distinctly below the positive control by the sols. Compared to the positive control, transcriptions of Cd transporter-related genes including OsLCT1, OsCCX2, OsHMA2 and OsPCR1 genes in leaves, and OsLCT1, OsCCX2, TaCNR2 and OSPCR1 in peduncles were downregulated by the increasing sols. Meanwhile, Se-binding protein 1 was evidently upregulated, together to retard Cd and Pb translocation to rice grains. The sols not only upregulated transcriptions of Lhcb1, RbcL, and OsBTF3 genes and production of psbA, Lhcb1 and RbcL proteins, but also increased the chlorophylls contents and RuBP carboxylase activities in the leaves, improving photosynthesis. The sols restrained ROS production from NADPH oxidases, but activated glutathione peroxidase, alleviating oxidative stress and damage. Additionally, Se was significantly enriched and was existed as selenomethionine in the rice grains. However, Pb transporter-related genes remain to be specified. Thus, the composite sols have potential to reduce Cd and Pb accumulation, mitigate oxidative damage, and promote photosynthesis and organic Se enrichment in rice plants under Cd and Pb combined pollution.

Large-scale survey of a neglected agent of sparganosis Spirometra erinaceieuropaei (Cestoda: Diphyllobothriidae) in wild frogs in China.

BACKGROUND: In China, frogs play an understudied role in the spread of human sparganosis (caused by the larval form of Spirometra). However, our knowledge about the prevalence of sparganum infection in frogs remains fragmented, and the taxonomic identification of the parasite is still controversial. METHODOLOGY/PRINCIPAL FINDINGS: The prevalence of sparganum infection in wild frogs was surveyed at 145 geographical locations from 28 of the 34 provinces/autonomous regions/municipalities in China for six years. The collected sparganum isolates from the different locations were subjected to molecular identification by a multiplex PCR assay and then were analysed with clustering analysis. In the survey, sparganum infection was found in 8 out of 13 of the collected frog species, and the most frequently infected species was Pelophylax nigromaculatus (the infection rate was up to 14.07%). Infected frogs were found in 80 of the 145 surveyed locations. The sparganum infection rates in the wild frogs in several regions of China were still high (above 10%), especially in South and Southwest China. A total of 72 spargana were selected for molecular identification, and the clustering analysis showed that sequences from the Chinese isolates were very similar to those identified as from Spirometra erinaceieuropaei. However, the taxonomy of the genus remains confused and further analysis is required. CONCLUSIONS: Eating wild frogs is associated with considerable health risks in China. Several traditional Chinese folk remedies may increase the risk of infection. The sparganum isolates in China are most likely from S. erinaceieuropaei, but new studies, especially comprehensive morphological analyses, are needed in the future.

[Effects of germplasm and host tree trunk on endophytic fungal communities in epiphytic Dendrobium catenatum].

In order to investigate the effects of germplasm and host tree trunk on endophytic fungal communities in epiphytic Dendrobium catenatum, a total of 3 835 isolates were recovered from roots, stems and leaves of four D. catenatum germplasms attached to one kind of host tree trunk and one germplasm attached to four kinds of epiphyte-host tree trunks. A total of 152 taxa were identified and classified based on the fungal cultural characteristics and phylogenetic analyses of ITS sequences. The taxa were assigned to 60 genera, 35 families, 21 orders and 5 classes of 2 phyla. The results indicated that D. catenatum cultivated in stereo cultivation harbor variety of fungi. The dominant fungal groups were different between Lin'an and Yiwu. Moreover, several groups showed geographical specificity, such as Arthrinium, Coniochaeta, Fusarium, Neofusicoccum and Zopfiella only dominating in Panshan of Lin'an, while Alternaria, Bjerkandera, Cercophora, Nigrospora and Trichoderma only dominating in Shangxi of Yiwu. There was no significant difference in diversity or species richness of endophytic fungi neither among germplasm nor host tree trunk. However, the richness and diversity indices exhibited a strong dependence on tissue type (P<0.05). The germplasm and host tree trunk impact the distribution patterns of endophytic fungi less than tissue type. Nevertheless, the relative frequencies of the dominant fungal groups were different among germplasms or host tree trunk types. Furthermore, there were some fungal species specific to certain germplasm or host tree trunk. This might be due to the distinctions in growth traits and chemical compositions of D. catenatum owning to the differences in D. catenatumgenetic background and microenvironment of host tree. Most of fungal taxa exhibit tissue specificity or preference. These results provide the basis for the study on the relationship between endophytic fungi and D. catenatum in stereo cultivation mode.

[Relationship between artesunate influence on the process of TGF-beta1 induced alveolar epithelial cells transform into mesenchymal cells and on idiopathic pulmonary fibrosis].

This study is to investigate the effect of artesunate on transforming growth factor-beta1 (TGF-beta1) induced epithelial-mesenchymal transition (EMT) and its possible mechanism. After the in vitro cultured RLE-6TN cells were treated with TGF-beta1 then artesunate intervened on it, after 24 h, expression of the markers of mesenchymal cell was assayed using Western blotting and real-time PCR analysis. Western blotting was also used to detect the effect of TGF-beta1 on the Smad3 and Smad7 expressions of RLE-6TN cells. Morphological alterations were examined by phase-contrast microscope, and ultrastructure changes by electron microscope. Incubation of RLE-6TN cells with TGF-beta1 resulted in the up-regulation of the expression of the mesenchymal cell markers, after artesunate intervened on it, resulted in the down-regulation of the expression. Meanwhile, incubation with artesunate intervened on RLE-6TN cells could lead to the apparent down-regulation of the expression of Smad3 and up-regulation of Samd7 and the transition of RLE-6TN cells to mesenchymal-like by TGF-beta1 induction, after artesunate intervened on it, RLE-6TN cells to epithelial-like. TGF-beta1 induced epithelial-mesenchymal transition process; artesunate can inhibit TGF-beta1-induced epithelial-mesenchymal transition process, the possible mechanism is up-regulation of the expression of Smad7 and down-regulation of the expression of Smad3, meanwhile inhibits phosphorylation of Smad3.

[Dendrobium officinale stereoscopic cultivation method].

The study is aimed to make the most of available space of Dendrobium officinale cultivation facility, reveal the yield and functional components variation of stereoscopic cultivated D. officinale, and improve quality, yield and efficiency. The agronomic traits and yield variation of stereoscopic cultivated D. officinale were studied by operating field experiment. The content of polysaccharide and extractum were determined by using phenol-sulfuric acid method and 2010 edition of "Chinese Pharmacopoeia" Appendix X A. The results showed that the land utilization of stereoscopic cultivated D. officinale increased 2.74 times, the stems, leaves and their total fresh or dry weight in unit area of stereoscopic cultivated D. officinale were all heavier than those of the ground cultivated ones. There was no significant difference in polysaccharide content between stereoscopic cultivation and ground cultivation. But the extractum content and total content of polysaccharide and extractum were significantly higher than those of the ground cultivated ones. In additional, the polysaccharide content and total content of polysaccharide and extractum from the top two levels of stereoscopic culture matrix were significantly higher than that of the ones from the other levels and ground cultivation. Steroscopic cultivation can effectively improves the utilization of space and yield, while the total content of polysaccharides and extractum were significantly higher than that of the ground cultivated ones. The significant difference in Dendrobium polysaccharides among the plants from different height of stereo- scopic culture matrix may be associated with light factor.

Relationship between artesunate influence on the process of TGF-beta1 induced alveolar epithelial cells transform into mesenchymal cells and on idiopathic pulmonary fibrosis / 药学学报

This study is to investigate the effect of artesunate on transforming growth factor-beta1 (TGF-beta1) induced epithelial-mesenchymal transition (EMT) and its possible mechanism. After the in vitro cultured RLE-6TN cells were treated with TGF-beta1 then artesunate intervened on it, after 24 h, expression of the markers of mesenchymal cell was assayed using Western blotting and real-time PCR analysis. Western blotting was also used to detect the effect of TGF-beta1 on the Smad3 and Smad7 expressions of RLE-6TN cells. Morphological alterations were examined by phase-contrast microscope, and ultrastructure changes by electron microscope. Incubation of RLE-6TN cells with TGF-beta1 resulted in the up-regulation of the expression of the mesenchymal cell markers, after artesunate intervened on it, resulted in the down-regulation of the expression. Meanwhile, incubation with artesunate intervened on RLE-6TN cells could lead to the apparent down-regulation of the expression of Smad3 and up-regulation of Samd7 and the transition of RLE-6TN cells to mesenchymal-like by TGF-beta1 induction, after artesunate intervened on it, RLE-6TN cells to epithelial-like. TGF-beta1 induced epithelial-mesenchymal transition process; artesunate can inhibit TGF-beta1-induced epithelial-mesenchymal transition process, the possible mechanism is up-regulation of the expression of Smad7 and down-regulation of the expression of Smad3, meanwhile inhibits phosphorylation of Smad3.

[Post-stroke shoulder-hand syndrome treated with acupuncture and rehabilitation: a randomized controlled trial].

OBJECTIVE: To assess the clinical efficacy on post-stroke shoulder-hand syndrome (SHS) treated with acupuncture and rehabilitation and the impacts on patients' nailfold microcirculation. METHODS: One hundred and twenty patients were randomized into an acupuncture rehabilitation group and a simple rehabilitation group, 60 cases in each one. In the simple rehabilitation group, OT (comprehensive rehabilitation therapy) training was adopted. In the acupuncture rehabilitation group, on the basis of the treatment as the simple rehabilitation, acu puncture was added at Taiyuan (LU 9), Zusanli (ST 36), Xuanzhong (GB 39), Waiguan (TE 5), Shousanli (LI 10), Quchi (LI 11) and Jianyu (LI 15). Acupuncture was given once a day, 7 days made one session. Totally, 4 sessions of treatment were required. Fugl-Meyer score, upper limb pain score, the score of nerve function defect and the items of nailfold microcirculation of patients were assessed in the the two groups before and after treatment. The efficacy was compared between the two groups. RESULTS: (1) The upper limb pain, the systematic motor function of the upper limbs, the nerve function defect, nailfold microcirculation and clinical symptoms were all improved after treatment in either the acupuncture rehabilitation group or the simple rehabilitation group as compared with those before treatment, indicating the significant difference (P<0. 05, P<0. 01). (2) The im provements in the upper limb pain (0. 90+/-0.71 vs 1. 80+/-0. 66), the systematic motor function of the upper limbs (42. 43 13. 57 vs 29. 98+/-15. 11), the nerve function defect (8. 60+/-11. 61 vs 13. 0+/-1. 74), nailfold microcirculation (total score 3. 18+/-1.32 vs 4.34+/-1.23) and clinical symptoms in the acupuncture rehabilitation group after treatment were different significantly as compared with those in the simple rehabilitation group (PO0. 05,P-O. 01), and the results in the acupuncture rehabilitation group were superior to the simple rehabilitation group. (3) In the acupuncture rehabilitation group, the markedly effective rate was 50. 0% (30/60) and the total effective rate was 93.3% (56/60), which was better than 16.7% (10/60) and 63. 3% (38/60) respectively in the simple rehabilitation group (all P<0. 05). CONCLUSION: Both the combined therapy of acupuncture and rehabilitation and the simple rehabilitation training are effective in the treatment of post-stroke SHS, and promote the status of nailfold microcirculation, the efficacy of the combined therapy is better than that of the latter.

[Establishment of high-throughput drug screening cell models based on JAK-STAT signal pathway].

AIM: To discover new drugs which may be applied to diseases of the immune system, hemogenesis system diseases and tumors, several high-throughput drug screening cell models based on JAK-STAT signal pathway have been established. METHODS: Four repeats of STAT DNA binding conserved sequences were synthesized, subcloned into pGL-Luc reporter vector and stably transfected into cell lines in vitro. Cell clones with high copy numbers of STAT binding sites and reporter genes were chosen as high-throughput drug screening cell models. The cell models were tested with known anti-allergic drugs and anti-tumor drugs by determining luciferase activity. The reaction was performed in 96 well micro-plates with a final volume of 50 microL. RESULTS: The cell models by performing rapid fluorescence assay were shown to be highly sensitive and stable after testing with cytokine and drugs. The modification of the expression plasmid simplified this method and made it more practical. It also provided good linear correlation, wide range of assay, highly sensitive and good reproducibility. CONCLUSION: The method can be performed by high-throughput drug screening for effective extraction of Chinese traditional herbs.

Establishment of high-throughput drug screening cell models based on JAK-STAT signal pathway / 药学学报

<p><b>AIM</b>To discover new drugs which may be applied to diseases of the immune system, hemogenesis system diseases and tumors, several high-throughput drug screening cell models based on JAK-STAT signal pathway have been established.</p><p><b>METHODS</b>Four repeats of STAT DNA binding conserved sequences were synthesized, subcloned into pGL-Luc reporter vector and stably transfected into cell lines in vitro. Cell clones with high copy numbers of STAT binding sites and reporter genes were chosen as high-throughput drug screening cell models. The cell models were tested with known anti-allergic drugs and anti-tumor drugs by determining luciferase activity. The reaction was performed in 96 well micro-plates with a final volume of 50 microL.</p><p><b>RESULTS</b>The cell models by performing rapid fluorescence assay were shown to be highly sensitive and stable after testing with cytokine and drugs. The modification of the expression plasmid simplified this method and made it more practical. It also provided good linear correlation, wide range of assay, highly sensitive and good reproducibility.</p><p><b>CONCLUSION</b>The method can be performed by high-throughput drug screening for effective extraction of Chinese traditional herbs.</p>