[Effect of electroacupuncture at "Zusanli" (ST 36) on duodenal mast cells, NGF and NTRK1 in rats with functional dyspepsia].

OBJECTIVE: To observe the effect of electroacupuncture (EA) at "Zusanli" (ST 36) on duodenal mast cells, nerve growth factor (NGF) and neurotrophic tyrosine kinase receptor type 1 (NTRK1), and to explore the mechanism of electroacupuncture at Zusanli (ST 36) on functional dyspepsia (FD). METHODS: Sixty SPF-grade 10-day-old SD rats were randomly divided into a normal group, a model group, a ketotifen group and an EA group, 15 rats in each group. The FD model was prepared by iodoacetamide combined with rat tail clamping method in the model group, the ketotifen group and the EA group. The rats in the ketotifen group were injected intraperitoneally with ketotifen (1 mg•kg-1•d-1) for 7 days; the rats in the EA group were treated with EA at bilateral "Zusanli" (ST 36), with disperse-dense wave, frequency of 2 Hz/50 Hz and intensity of 0.5 mA, 20 min each time, once a day for 14 days. The gastric emptying rate and small intestinal propulsion rate in each group were observed; the morphology of duodenal mucosa was observed by HE staining; the toluidine blue staining was used to observe the number and degranulation of mast cells in duodenal mucosa; the protein and mRNA expressions of NGF, NTRK1 in duodenum were detected by Western blot and real-time PCR; the level of interleukin-1ß (IL-1ß) in duodenum was measured by ELISA. RESULTS: Compared with the normal group, the gastric emptying rate and small intestinal propulsion rate in the model group were decreased (P<0.01); compared with the model group, the gastric emptying rate and small intestinal propulsion rate in the ketotifen group and the EA group were increased (P<0.01); the small intestinal propulsion rate in the EA group was higher than that in the ketotifen group (P<0.01). In the model group, local defects in duodenal mucosa were observed with a small amount of inflammatory cell infiltration; no obvious abnormality was found in duodenal mucosa of the other groups. Compared with the normal group, the mast cells of duodenal mucosa in the model group were increased significantly with significant degranulation; compared with the model group, the mast cells of duodenal mucosa in the ketotifen group and the EA group were decreased significantly, and the degranulation was not obvious. Compared with the normal group, the protein and mRNA expressions of NGF, NTRK1 as well as the level of IL-1ß in duodenum in the model group were increased (P<0.01); compared with the model group, the protein and mRNA expressions of NGF, NTRK1 as well as the levels of IL-1ß in duodenum in the ketotifen group and the EA group were decreased (P<0.01, P<0.05); compared with the ketotifen group, the mRNA expression of NGF, as well as the protein and mRNA expressions of NTRK1 in duodenum in the EA group were decreased (P<0.05, P<0.01). CONCLUSION: EA at "Zusanli" (ST 36) could inhibit the activation of duodenal mast cells and regulate the expressions of NGF and its receptor to improve the low-grade inflammatory response of duodenum, resulting in treatment effect on FD.

[Influence of electroacupuncture of"Zusanli"(ST36)on mast cells/TRPV1 signaling pathway in visceral hypersensitivity rats with functional dyspepsia].

OBJECTIVE: To explore the interventional mechanism of electroacupuncture (EA) of "Zusanli"(ST36)based on the involvement of mast cells/ transient receptor potential vanilloid type1 (TRPV1) signaling pathway in relieving visceral hypersensitivity in functional dyspepsia (FD) rats. METHODS: Sixty SD rats (half male and half female, 10 days in age) were randomly divided into normal control, model, medication (ketotifen) and EA groups, with 15 rats in each group. The FD model was established by gavage of iodoacetamide combined with tail clamping (stress stimulation). Rats of the medication group received intraperitoneal injection of ketotifen (1 mg·kg-1·d-1) for 14 d, and those of the EA group received EA of ST36 for 20 min, once a day for 14 d. An air-balloon was inserted into the rat's stomach for recording changes of the intragastric pressure (mL/mm Hg) via a pressure transducer. The visceral hypersensitivity was assessed using abdominal withdrawal reflex (AWR) score and the number and degranulation of mast cells of gastric mucosa were observed using toluidine blue staining. The expression levels of TRPV1 and proteinase activated receptor 2 (PAR2) in the stomach were observed using immunofluorescence histochemistry and Western blot, separately, and the contents of SP and CGRP in the stomach detected using ELISA. RESULTS: When the intragastric pressure was at 50, 60 and 70 mm Hg, the gastric compliance was significantly decreased (P<0.01), and the levels of visceral sensitivity increased in the model group （P<0.01）。 TRPV1 immunofluorescence tensity, expression of PAR2 and TRPV1 proteins, and contents of SP and CGRP in the stomach were considerably up-regulated in the model group compared with the normal control group (P<0.01). In comparison with the model group, under intragastric pressure of 50，60 and 70 mm Hg, the gastric compliance was obviously increased, and the visceral hypersensitivity decreased in the EA group （P<0.01,P<0.05）. TRPV1 immunofluorescence intensity, expression levels of PAR2 and TRPV1 proteins, and the contents of SP and CGRP in the stomach were considerably down-regulated in both medication and EA groups compared with the model group (P<0.01, P<0.05). The therapeutic effect of EA was significantly superior to that of medication in up-regulating the gastric compliance （at 70 mm Hg）, and down-regulating the contents of SP and CGRP (P<0.05). No significant differences were found between the EA and medication groups in up-regulating gastric compliance at intragastric pressure of 50 and 60 mm Hg, and in down-regulating the visceral sensitivity, TRPV1 fluorescence intensity, and expression of PAR2 and TRPV1 proteins (P>0.05). Toluidine blue staining showed an apparent increase of mast cell number with obvious degranulation in the gastric mucosa of rats in the model group, which was milder in the EA and medication groups. CONCLUSION: EA of ST36 can suppress visceral hypersensitivity and increase the gastric compliance in FD rats, which may be related with its effects in inhibiting the activation of gastric mast cells, and down-regulating the expression of gastric PAR2 and TRPV1 proteins and SP and CGRP contents.

Chinese medicine Yu-Ping-Feng-San attenuates allergic inflammation by regulating epithelial derived pro-allergic cytokines.

This study aimed to investigate the mechanisms of Yu-Ping-Feng-San (YPFS) on attenuating allergic inflammation in the initial stage of atopic dermatitis (AD). AD mouse model was established with fluorescein isothiocyanate (FITC) sensitization and elicitation. Epithelial barrier structure was observed with transmission electron microscope. The populations of dendritic cells (DCs) and group 2 innate lymphoid cells (ILC2s) were detected by flow cytometry. Human immortalized keratinocyte (HaCaT) cells were stimulated with Poly(I:C)/TNF-α in vitro to assessthymic stromal lymphopoietin (TSLP), interleukin (IL)-33 and nuclear factor-κB (NF-κB) levels or expressions by immunofluorescence, enzyme linked immunosorbent assay (ELISA) and western blot. In the initial stage of AD, ear swelling and infiltration of inflammatory cells in ear tissues were markedly attenuated with YPFS treatments. The damaged structures of ear epithelium and the increased levels of Th2-cytokines induced by FITC were significantly rescued in YPFS-treated mice. The production of pro-allergic cytokines, TSLP and IL-33, as well as the cell populations of their target cells DCs and ILC2s were decreased in AD model, respectively. Likewise, the levels of TSLP and IL-33 in Poly(I:C)/TNF-α-stimulated HaCaT cells showed the same results. Lower levels of p-NF-κB were detected with YPFS treatment, and the expressions of TSLP and IL-33 could be further decreased with inhibiting of NF-κB. Therefore, YPFS attenuates allergic inflammation in the initial stage of AD probably through regulating NF-κB-TSLP/IL-33 pathway, which may provide a novel effective target for the prevention and treatment of allergic diseases.

A new sesquiterpene lactone from yacon leaves.

The chemical constituents of 60% EtOH extract of yacon leaves were separated to yield a new compound, together with four known compounds, which were isolated for the first time from yacon. The new compound was characterised and named as chlorodalin (1) on the basis of NMR (1D and 2D), HR-MS and other spectral methods. The cytotoxic activities of 1-5 were evaluated on two human tumour cell lines and the new compound showed significant cytotoxic activity.

Molecular Cloning and Characterization of Full-Length cDNA of Calmodulin Gene from Pacific Oyster Crassostrea gigas.

The shell of the pearl oyster (Pinctada fucata) mainly comprises aragonite whereas that of the Pacific oyster (Crassostrea gigas) is mainly calcite, thereby suggesting the different mechanisms of shell formation between above two mollusks. Calmodulin (CaM) is an important gene for regulating the uptake, transport, and secretion of calcium during the process of shell formation in pearl oyster. It is interesting to characterize the CaM in oysters, which could facilitate the understanding of the different shell formation mechanisms among mollusks. We cloned the full-length cDNA of Pacific oyster CaM (cgCaM) and found that the cgCaM ORF encoded a peptide of 113 amino acids containing three EF-hand calcium-binding domains, its expression level was highest in the mantle, hinting that the cgCaM gene is probably involved in shell formation of Pacific oyster, and the common ancestor of Gastropoda and Bivalvia may possess at least three CaM genes. We also found that the numbers of some EF hand family members in highly calcified species were higher than those in lowly calcified species and the numbers of these motifs in oyster genome were the highest among the mollusk species with whole genome sequence, further hinting the correlation between CaM and biomineralization.

A Case Report of Locked-in Syndrome Due to Bilateral Vertebral Artery Dissection After Cervical Spine Manipulation Treated by Arterial Embolectomy.

Cervical spine manipulation (CSM) is a commonly spinal manipulative therapies for the relief of cervical spine-related conditions worldwide, but its use remains controversial. CSM may carry the potential for serious neurovascular complications, primarily due to vertebral artery dissection (VAD) and subsequent vertebrobasilar stroke. Here, we reported a rare case of locked-in syndrome (LIS) due to bilaterial VAD after CSM treated by arterial embolectomy.A 36-year-old right-handed man was admitted to our hospital with numbness and weakness of limbs after treating with CSM for neck for half an hour. Gradually, although the patient remained conscious, he could not speak but could communicate with the surrounding by blinking or moving his eyes, and turned to complete quadriplegia, complete facial and bulbar palsy, dyspnea at 4 hours after admission. He was diagnosed with LIS. Then, the patient was received cervical and brain computed tomography angiography that showed bilateral VAD. Aortocranial digital subtraction angiography showed vertebrobasilar thrombosis, blocking left vertebral artery, and stenosis of right vertebral artery. The patient was treated by using emergency arterial embolectomy and followed by antiplatelet therapy and supportive therapy in the intensive care unit and a general ward. Twenty-seven days later, the patient's physical function gradually improved and discharged but still left neurological deficit with muscle strength grade 3/5 and hyperreflexia of limbs.Our findings suggested that CSM might have potential severe side-effect like LIS due to bilaterial VAD, and arterial embolectomy is an important treatment choice. The practitioner must be aware of this complication and should give the patients informed consent to CSM, although not all stroke cases temporally related to SCM have pre-existing craniocervical artery dissection.

[Effect of Modified Baizhu Powder on Gastrointestinal Functions in Mouse Models with Stomach-cold Functional Dyspepsia].

OBJECTIVE: To explore the effect of modified Baizhu (Rhizoma Atractylodis Macrocephalae) powder on the gastrointestinal function in mouse models with stomach-cold functional dyspepsia. Meanwhile,the mouse models were administered with Shihu (dendrobium), a traditional Chinese drug with cold nature and flavour, to explore the way via which it exert its effect on specific symptoms. Methods: Mouse models with stomach-cold functional dyspepsia were established by ice water and ice NaOH. The effects of modified Baizhu powder and dendrobium on mice were observed in terms of water intake, weight change,small intestine propulsion rate, intestinal absorption function, and effects on ghrelin and motilin. RESULTS: The modified Baizhu powder effectively increased food intake, water intake, body weight (P<0.05) and swimming time (P<0.01), increased the small intestine propulsion rate and serum D-xylose content (P<0.05), and up-regulated ghrelin (P<0.05). Also, it showed a trend to down-regulate the motilin, although the change was not statistically significant (P>0.05). In contrast,the use of Shihu aggravated symptoms in the mouse models. Conclusion: The changes in ghrelin and motilin levels may be the neuro-endocrine mechanisms via which the modified Baizhu powder and Shihu exert their effects on mouse models.

[Influence of feeding patterns on the development of teeth, dentition and jaw in children].

Breastfeeding has been recognized as the most natural and nutritious way of feeding babies. Besides the nutritional, immunological and emotional benefits, breastfeeding promotes a healthy stomatognathic system. First of all, the nutrients and minerals in maternal milk are easy to be absorbed by the infants, which contributes to the mineralization of the teeth, and suppress the propagation of bacteria on the teeth. Though the jury is still out on whether breastfeeding can prevent Early Childhood Caries (ECC), it is definite that we should pay attention to feeding at night and the oral hygiene of the babies. Secondly, the method of feeding is closely bound up with the development of dentition and jaw. Breast- and bottle-feeding involve different orofacial muscles, which possibly have different effects on the harmonic growth of maxilla and dental arches. Meanwhile, breathing, swallowing and mastication should be developing in harmony, and differences exist in the learning of the coordinated movement between breast feeding and bottle feeding children. Bottle feeding had been proved to be closely related with the non-nutritive sucking habits which can cause malocclusion. At last, it should be pointed out that breast feeding should be the only feeding source in the first 6 months of life, then supplementary foods should be added. And prolonged bottle feeding should be avoided. We can see that breast feeding is definitely good for the infants, but the reality is not optimistic in our country.

Protective effect of Dl-3n-butylphthalide on learning and memory impairment induced by chronic intermittent hypoxia-hypercapnia exposure.

Cognitive impairment is a common finding in patients with chronic obstructive pulmonary disease (COPD), but little attention has been focused on therapeutic intervention for this complication. Chronic intermittent hypoxia hypercapnia (CIHH) exposure is considered to be responsible for the pathogenesis of COPD. Dl-3n-Butylphthalide (NBP), extracted from Apium graveolens Linn, has displayed a broad spectrum of neuroprotective properties. Our study aimed to investigate the potential of NBP on CIHH-induced cognitive deficits. The cognitive function of rats after CIHH exposure was evaluated by the Morris water maze, which showed that the NBP treated group performed better in the navigation test. NBP activated BDNF and phosphorylated CREB, the both are responsible for neuroprotection. Additionally, NBP decreased CIHH induced apoptosis. Moreover, NBP further induced the expression of HIF-1α, accompanied by the up-regulation of the autophagy proteins Bnip3, Beclin-1 and LC3-II. Finally, NBP also reversed the decreased expression of SIRT1 and PGC-1α, but the expression of Tfam, Cox II and mtDNA remained unchanged. These results suggested that the neuroprotective effects of NBP under CIHH condition possibly occurred through the inhibition of apoptosis, promotion of hypoxia-induced autophagy, and activation of the SIRT1/PGC-1α signalling pathway, while stimulation of mitochondrial biogenesis may not be a characteristic response.

Epidemiology of complementary and alternative medicine use in patients with Parkinson's disease.

Parkinson's disease (PD) is a common and debilitating neurodegenerative disorder without a known neuroprotective cure. Currently, an increasing number of patients with PD resort to complementary and alternative medicine (CAM). This study aimed to determine the epidemiology of CAM use for PD worldwide. Methodological issues included the definition of CAM, running a search strategy using five databases, and citation tracking. Six studies estimated the prevalence of CAM use for PD to be between 25.7% and 76%. The response rates in these surveys varied from 81% to 100%. Frequently utilized forms of therapy were acupuncture, massage, herbs, and vitamins/health supplements, and these therapies were mainly used to improve the associated motor symptoms of PD. However, only 11% to 20% of these patients were referred to use CAM by a healthcare professional. Of the sociodemographic and disease-specific factors, CAM use was correlated with female sex, age, age at onset of PD, longer duration of PD, degree of education, higher income, rural location, comorbidity for indications, levodopa load, and severe motor symptoms. These results suggested that CAM use is widespread among patients with PD worldwide, but the largely unexamined use of CAM requires more attention. Moreover, there is a lack of communication between physicians and patients, increasing the risks associated with CAM use and the potential for adverse events.

Cytotoxic constituents from the leaves of Broussonetia papyrifera.

AIM: To investigate the chemical constituents from the leaves of Broussonetia papyrifera. METHODS: The chemical constituents were isolated and purified by macroporous adsorptive resin D101, silica gel, and ODS column chromatography and preparative HPLC. Their structures were elucidated on the basis of 1D and 2D NMR analyses. In addition, their cytotoxic activity against human hepatoma carcinoma cells (HepG-2) were evaluated by the MTT method. Furthermore, RP-HPLC and colorimetric methods were used for the analysis of cosmosiin and total flavonoids. RESULTS: A new lignan, together with five known compounds were obtained, and their structures were characterized as (+)-pinoresinol-4'-O-ß-D-glucopyranosyl-4″-O-ß-D-apiofuranoside (1), cosmosiin (2), luteolin-7-O-ß-D-glucopyranoside (3), liriodendrin (4), 3, 5, 4'-trihydroxy-bibenzyl-3-O-ß-D-glucoside (5), and apigenin-6-C-ß-D-glucopyranside (6). Furthermore, RP-HPLC and colorimetric methods were established for the analysis of cosmosiin and total flavonoids. CONCLUSION: Compound 1 was a new lignan, and compounds 5 and 6 were isolated for the first time from the title plant. Compounds 1, 4 and 6 showed definite activities against HepG-2, while the other compounds didn't show inhibitory effects. The optimal harvest time of B. papyrifera (L.) Vent. is September.

Clinical efficacy and safety of Chinese herbal medicine for Wilson's disease: a systematic review of 9 randomized controlled trials.

Wilson's disease is an autosomal recessive disorder of copper metabolism. Despite being treatable, there is no universally accepted treatment regimen. Currently, various Chinese herbal medicines (CHMs) are widely used in the treatment of Wilson's disease in China, but there is a lack of reliable scientific evidence for the effectiveness of such therapies. The objective of this systematic review is to assess the clinical efficacy and safety of CHM as an alternative or/and adjuvant therapy for Wilson's disease. A systematic literature search in different medical databases was performed to identify randomized controlled trials comparing CHM as monotherapy or CHM as adjuvant therapy with western conventional medical therapy in the treatment of Wilson's disease. A total of 687 participants were included in nine eligible studies. The main findings are that CHM as monotherapy or adjuvant therapy for Wilson's disease may be able to improve the clinical symptoms, to promote the urinary copper excretion, to ameliorate liver function and/or liver cirrhosis, and has fewer adverse effects in comparison with western conventional medication. Furthermore, CHM generally appeared to be safe and well tolerated in patients with Wilson's disease. However, the evidence presented in this review are insufficient to warrant a clinical recommendation due to the generally low methodological quality of the included studies. In conclusion, CHM seems to be beneficial and safe for Wilson's disease, but high-quality evidences are still needed to further evaluate this therapy. Therefore, additional well-designed, randomized, placebo-controlled clinical trials are needed.

[Effects of 5-FU combined compound ginseng and astragalus on biological behavior of human gastric cancer MGC-803 cells].

OBJECTIVE: To observe the in vitro effects of 5-fluorouracil (5-FU) combined Compound Ginseng and Astragalus (CGA) on the biological behaviors such as the proliferation, the cloning, apoptosis and migration of human gastric cancer MGC-803 cells. METHODS: The cell proliferation inhibition rate was detected by MTT assay, and the median effective concentrations of different drugs used alone/combination were calculated. The cell cycle and apoptosis were observed by flow cytometry. The formation of the cell colony was detected by Giemsa staining. The drugs' inhibition on cell migration was detected by cell scratch experiment. RESULTS: CGA and 5-FU both could inhibit the growth of the human gastric cancer cell line MGC-803 cells, and their effects were enhanced along with increased drug concentrations. Compared with CGA or 5-FU alone, CGA +5-FU got higher cell growth inhibition rate (P<0.05), and the effects were enhanced along with increased concentrations. CGA and 5-FU both could induce the apoptosis of MGC-803 cells, inhibit the formation of cell cloning, block cells at G0/G1 phase, and inhibit the cell migration. Compared with CGA or 5-FU alone, CGA + 5-FU got higher apoptosis rate of MGC- 803 cells, and more cells blocked at G0/G1 phase (P<0.05). Besides, the MGC-803 cells were inhibited at G0/G1 phase. Compared with CGA or 5-FU alone, CGA +5-FU obviously lowered formation of cell cloning and area of cell migration (P<0.05). The median effective concentration of CGA +5-FU was less than the sum of the median effective concentration of CGA and 5-FU. CONCLUSION: Compared with CGA or 5-FU alone, CGA +5-FU could better inhibit the cell growth of human gastric cancer MGC- 803 cells, suppress the formation of cell cloning, induce cell apoptosis, block the cell cycle at G0/G1 phase, and inhibit the cell migration.

Ginseng total saponins enhance neurogenesis after focal cerebral ischemia.

ETHNOPHARMACOLOGICAL RELEVANCE: Ginseng, the root of Panax ginseng C.A. Meyer, is one of the most commonly used healing herbs for stroke and chronic debilitating conditions in China. Ginsenosides are the main active principles for ginseng's efficacy, but the mechanisms have not been fully clarified. AIM OF THE STUDY: To test the hypothesis whether or not the administration of Ginseng total saponins (GTS) can enhance neurogenesis after focal cerebral ischemia, and thereby improve neurological deficits. MATERIALS AND METHODS: Male Wistar rats received intraperitoneal injections of GTS dissolved at a dose of 25 mg kg(-1) d(-1) or normal saline (NS) of same volume 3 days before the permanent middle cerebral artery occlusion (MCAO) model establishment until the animals were killed at the time points of 1d, 3d, 7d and 14d. The neurological function was assessed blindly. BrdU immunostaining and double staining were performed by following the 3-steps method. RESULTS: (A) GTS-treated rats have better neurological scores compared with those in NS group at 14d time point (p<0.05); (B) the number of BrdU(+) cells and BrdU(+)/NeuN(+) cells in GTS group were significantly higher than those in NS group in the ipsilateral subventricular zone and in the ipsilateral infarct area after MCAO, respectively (p<0.05 or p<0.01); (C) the increase of the number of BrdU(+)/NeuN(+) cells highly correlated with the decrease of neurological scores. Coefficient correlation r=-0.828 (p<0.01). CONCLUSION: GTS can improve neurological deficits after focal cerebral ischemia by inducing endogenous neural stem cells activation and thereby enhance adult central nervous system regeneration.

[Establishment and development of Xuan Fu (sweat pore) Theory.]

Xuan Fu (sweat pore) theory was derived from Huangdi Nei Jing (Canon of Internal Medicine), formulated by Liu Wan-su in his Su Wen Xuan Ji Yuan Bing Shi (Pattern of Mysterious Mechanism of Pathogenesis in Plain Questions) in the Jin Dynasty, and perfected by doctors later. The article introduces the establishment and development of Xuan Fu (sweat pore) theory from its concept and function, pathology for its stasis, therapeutical principles and treatment for opening Xuan Fu, its special theory and modern exploration in its essence.