[Acupoint selection rules of neurogenic dysphagia treated with acupuncture and moxibustion in ancient times].

Acupoint selection rules of neurogenic dysphagia treated with acupuncture and moxibustion from pre-Qin to late Qing Dynasty in Chinese Medical Classics were analyzed based on data mining. The literature regarding acupuncture and moxibustion for neurogenic dysphagia was searched and screened according to the inclusion and exclusion criteria in Chinese Medical Classics (5th Edition), the prescriptions were extracted according to the principle of acupoint extraction.The SPSS 21.0 and Clementine 12.0 were used to perform the cluster analysis and association rule analysis.A total of 191 acupuncture and moxibustion prescriptions were screened and extracted，including 45 acupoints. The top 5 acupoints of acupuncture and moxibustion for neurogenic dysphagia in frequency were Jiache (ST 6), Dicang (ST 4), Lieque (LU 7), Lianquan (CV 23), Shuigou (GV 26). The most involved meridians were the stomach meridian, the governor vessel and the conception vessel. The main acupoints were distributed in the scalp face neck and upper limbs. The most frequently used specific acupoints was crossing points,next was eight confluence points. There were 11 acupoint combinations with strong association according to the association rule analysis, and the top one acupoint combination was Hegu (LI 4)-Jiache (ST 6). There were 9 acupoint cluster groups according to the cluster analysis. Hegu (LI 4), Jiache (ST 6), Dicang (ST 4), Shuigou (GV 26) and Chengjiang (CV 24) were core acupoints for neurogenic dysphagia treated with acupuncture and moxibustion in ancient times, besides，selecting proximal and distal acupoints and selecting acupoints according to symptoms were emphasized.

[Acupuncture for dysarthria: systematic review].

OBJECTIVE: To systematically evaluate the clinical efficacy of acupuncture for dysarthria, and to explore the rules of acupoints selection for dysarthria. METHODS: The clinical randomized control trial literature regarding acupuncture for dysarthria published before January of 2018 were searched in databases, including CNKI, Wanfang, VIP, CBM, PubMed, Ebsco, Science Direct and Cochrane Library. The information of included studies was extract and the quality was assessed. The Meta analysis was performed by using RevMan 5.3 software. The frequency of acupoints was calculated by using Excel software to analyzed the rules of acupoints selection. RESULTS: Totally 21 papers were included, involving 1651 patients. The pooled effects of clinical efficacy: heterogeneity test P =0.74, I 2=0%, OR =6.36, 95% CI: 4.55, 8.88, Z =10.84 (P<0.01), indicating the efficacy in the treatment group was significantly higher than that in the control group. The pooled effects of the symptom score in Frenchay scale: heterogeneity test P =0.56, I 2=0%, WMD =3.20, 95% CI: 1.38, 5.02, Z =3.45 (P<0.01), indicating the efficacy in the treatment group was significantly higher than that in the control group. The acupoints with frequency of more than 5 times were Fengchi (GB 20), Yuye (EX-HN 13), Jinjin (EX-HN 12), Lianquan (CV 23), Baihui (GV 20), tongue-three needles and Yamen (GV15). The meridians with frequency of more than 5 times were the extra channels, governor vessel, gallbladder channel, conception vessel and stomach channel. CONCLUSION: The clinical efficacy of acupuncture combined with speech training/regular treatment is significantly superior to that of control group (speech training, medication, regular treatment); acupuncture is safe and effective for dysarthria; the majority of selected acupoint is local acupoints around tongue, throat and neck, as well as extra points and empirical points. However, high-quality randomized controlled trials with large sample sizes are still needed to provide further evidence.

Membrane lipid composition regulates tubulin interaction with mitochondrial voltage-dependent anion channel.

Elucidating molecular mechanisms by which lipids regulate protein function within biological membranes is critical for understanding the many cellular processes. Recently, we have found that dimeric αß-tubulin, a subunit of microtubules, regulates mitochondrial respiration by blocking the voltage-dependent anion channel (VDAC) of mitochondrial outer membrane. Here, we show that the mechanism of VDAC blockage by tubulin involves tubulin interaction with the membrane as a critical step. The on-rate of the blockage varies up to 100-fold depending on the particular lipid composition used for bilayer formation in reconstitution experiments and increases with the increasing content of dioleoylphosphatidylethanolamine (DOPE) in dioleoylphosphatidylcholine (DOPC) bilayers. At physiologically low salt concentrations, the on-rate is decreased by the charged lipid. The off-rate of VDAC blockage by tubulin does not depend on the lipid composition. Using confocal fluorescence microscopy, we compared tubulin binding to the membranes of giant unilamellar vesicles (GUVs) made from DOPC and DOPC/DOPE mixtures. We found that detectable binding of the fluorescently labeled dimeric tubulin to GUV membranes requires the presence of DOPE. We propose that prior to the characteristic blockage of VDAC, tubulin first binds to the membrane in a lipid-dependent manner. We thus reveal a new potent regulatory role of the mitochondrial lipids in control of the mitochondrial outer membrane permeability and hence mitochondrial respiration through tuning VDAC sensitivity to blockage by tubulin. More generally, our findings give an example of the lipid-controlled protein-protein interaction where the choice of lipid species is able to change the equilibrium binding constant by orders of magnitude.

Bacteria-induced static batch fungal fermentation of the diterpenoid cyathin A(3), a small-molecule inducer of nerve growth factor.

Cyathin A(3), produced by the fungus Cyathus helenae, is a member of the cyathane family of diterpene natural products. While many of the cyathanes display antibacterial/antimicrobial activity or have cytotoxic activity against human cancer cell lines, their most exciting therapeutic potential is derived from their ability to induce nerve growth factor (NGF) release from glial cells, making the cyathanes attractive lead molecules for the development of neuroprotective therapeutics to prevent/treat Alzheimer's disease. To investigate if cyathin A(3) has NGF-inducing activity, we set out to obtain it using published C. helenae bench-scale fungal fermentations. However, to overcome nonproducing fermentations, we developed an alternative, bacteria-induced static batch fermentation approach to the production of cyathin A(3), as described in this report. HPLC, UV absorption spectra, and mass spectrometry identify cyathin A(3) in fungal fermentations induced by the timely addition of Escherichia coli K12 or Bacillus megabacterium. Pre-filtration of the bacterial culture abolishes cyathin A(3) induction, suggesting that bacteria-associated media changes or physical interaction between the fungus and bacteria underlie the induction mechanism. Through alteration of incubation conditions, including agitation, the timing of induction, and media composition, we optimized the fermentation to yield nearly 1 mg cyathin A(3)/ml media, a sixfold increase over previously described yields. Additionally, by comparison of fermentation profiles, we reveal that cyathin A(3) biosynthesis is regulated by carbon catabolite repression. We have used an enzyme-linked immunosorbent assay to illustrate that cyathin A(3) induces NGF release from cultured glial cells, and therefore cyathin A(3) warrants further examination in the development of neuroprotective therapeutics.