[Research progress of the impacts of acupuncture on programmed cell death in treatment of ischemic stroke].

The new progress has been made in the research of programmed cell death (e.g. autophagy, apoptosis, pyroptosis, necroptosis and ferroptosis) for the pathological mechanism of ischemic stroke. As an important non-pharmacological therapy, acupuncture is widely used in stroke patients and has achieved favorable effect. The researches in recent years have shown that acupuncture plays its neuroprotective role on ischemic stroke by modulation of autophagy, apoptosis, pyroptosis, necroptosis and ferroptosis of neurons. Acupuncture is effective in treatment of ischemic stroke by regulating programmed cell death.

An miR156-regulated nucleobase-ascorbate transporter 2 confers cadmium tolerance via enhanced anti-oxidative capacity in barley.

INTRODUCTION: Cadmium (Cd) is one of the most detrimental heavy metal pollutants, seriously affecting crop production and human health. Nucleobase-ascorbic acid transporters (NAT) are widely present in many living organisms including plants, animals and microbes; however, the role of NAT in plant Cd tolerance remains unknown. OBJECTIVES: To identify Cd-induced miRNAs that target HvNAT2 and to determine the role of this gene and its product in Cd tolerance. METHODS: High-throughput-sequencing was used to identify the miRNA expression profile of barley roots in response to Cd stress. Overexpression (OX) and RNAi lines were then constructed for HvNAT2 and comparative transcriptomic analysis was performed to determine the function of this transporter examining its effects on traits such as Cd uptake/flux and translocation, morphology and antioxidant capacity in relation to Cd tolerance. In addition, phylogenetic analysis was performed to obtain insights into the evolution of HvNAT2. RESULTS: Cd stress-induced genome-wide expression profiles of miRNAs identified a Cd-induced miRNA, miR156g-3p_3, that had HvNAT2 as its target. HvNAT2 was negatively regulated in the high-Cd-accumulating and Cd-tolerant genotype Zhenong8. Evolutionary analysis indicated that orthologues of the plasma membrane localized, HvNAT2, can be traced back to the sister group of land plants, the streptophyte algae. Overexpression of HvNAT2 increases Cd tolerance with higher tissue Cd accumulation but less oxidative damage in transgenic barley plants. RNAi of HvNAT2 leads to a significant reduction of Cd tolerance. The higher Cd accumulation in roots of the OX3 line was also demonstrated by confocal microscopy and electrophysiology. Transcriptome analysis showed that the enhancement of antioxidant capacity by HvNAT2 was related to stress signaling pathways. Furthermore, oxidative stress tolerance in HvNAT2-OX plants was regulated by the synthesis of phytochelatins and the glutathione metabolism cycle. CONCLUSION: Our study reveals a key molecular mechanism of NAT in Cd tolerance in plants that is useful for sustainable agricultural production and management of hazardous this heavy metal for better environment management and ecosystem function.

EGFR-mediated G1/S transition contributes to the multidrug resistance in breast cancer cells.

Despite the improvement of strategies against cancer therapy, the multidrug resistance (MDR)is the critical problem for successful cancer therapy. Recurrent cancers after initial treatment with chemotherapy are generally refractory to second treatments with these anticancer therapies. Therefore, it is necessary to elucidate the therapy-resistant mechanism for development of effective therapeutic modalities against tumors. Here we demonstrate a phase-specific chemotherapy resistance due to epidermal growth factor receptor (EGFR) in human breast cancer cells. Thymidine-induced G1-arrested cultures showed upregulated chemosensitivity, whereas S-phase arrested cells were more resistant to chemotherapeutic agents. Overexpression of EGFR promoted the MDR phenotypes in breast cancer cells via accelerating the G1/S phase transition, whereas depletion of EGFR exerted the opposite effects. Furthermore, CyclinD1, a protein related to cell cycle, was demonstrated to be involved in above EGFR-mediated effects since EGFR increased the expression of CyclinD1, and the specific RNA interference against CyclinD1 could primarily abolish the EGFR-induced MDR phenotypes. These data provide new insights into the mode by which MDR breast cancers evade cytoxic attacks from chemotherapeutic agents and also suggest a role for EGFR-CyclinD1 axis in this process.

[Effects of early application of Tuina treatment on quadriceps surface myoelectricity in patients after total knee arthroplasty: a randomized controlled trial].

BACKGROUND: Total knee replacement surgery is commonly used in end-stage diseases of the knee. It is important for improving surgical efficacy and patient satisfaction by promoting early rehabilitation of patients and improving knee function. OBJECTIVE: To observe the effects of early application of Tuina treatment on quadriceps surface electromyography (EMG) in patients with rheumatoid arthritis having undergone total knee arthroplasty. DESIGN, SETTING, PARTICIPANTS AND INTERVENTION: The study was performed at the Orthopedic Department of Huashan Hospital, Fudan University, and the Shanghai Guanghua Hospital of Integrated Traditional Chinese and Western Medicine from June 2010 to September 2011. A total of 66 patients with rheumatoid arthritis who had undergone total knee replacement surgery were randomly divided into control group and observation group, 33 cases in each. The patients in the control group were administered with continuous passive training (CPM), and the patients in the observation group were treated with CPM combined with Tuina, from prior surgery to four weeks post-surgery. MAIN OUTCOME MEASURES: The knee function was evaluated using the Western Ontario and McMaster Universities Osteoarthritis Index (WOMAC) questionnaire at baseline and 4 weeks after the surgery. Quadriceps surface EMG was also detected at the same time points. RESULTS: After 4 weeks of Tuina and comprehensive rehabilitation intervention, the WOMAC questionnaire score of the observation group was decreased compared with the control group (P<0.01); median frequency and integrated electromyography of the rectus femoris and vastus medialis muscles, which were recorded by EMG, in the observation group were higher than those in the control group (P<0.01). CONCLUSION: Tuina can improve the recovery of patients who have undergone total knee replacement by increasing quadriceps EMG.

[Effect of electroacupuncture invention on activities of PGC-1α/Irisin (FNDC5)/BDNF signaling in cerebral cortex, hippocampus and muscles in focal cerebral ischemic/reperfusion injury rats].

OBJECTIVE: To observe the effect of electroacupuncture(EA) on the expression of peroxisome proliferator-activated receptor gamma coactivators-1-alpha (PGC-1α), Irisin and brain-derived neurotrophic factor (BDNF) in the ischemic peripheral cortex, hippocampus and local skeletal muscle in rats with focal cerebral ischemic/reperfusion injury (CI/RI), so as to explore its underlying mechanism of improving of CI/RI. METHODS: Male SD rats were randomly divided into 3 groups: sham-operation, model and EA (11 rats in each group). The focal CI/RI model was established by middle cerebral artery occlusion (MCAO). EA (2 Hz /15 Hz, 2 to 4 mA) was applied to "Quchi" (LI11) and "Zusanli" (ST36) of the affected side for 20 min, once a day for 7 days. Zea-Longa's score and Balance Beam score were used to evaluate the neurological and motor functions. The infarcted volume of the brain was detected by using 2,3,5-triphenyltetrazolium chloride staining. The expression levels of PGC-1α, fibronectin type III domain-containing protein 5(FNDC5) and BDNF proteins in the ischemic peripheral cortex, hippocampus and local skeletal muscle were detected by Western blot. RESULTS: Compared with the sham-operation group, the Zea-Longa's score, Balance Beam score, percentage of cerebral infarct volume were notably increased (P<0.01), while the expression levels of PGC-1α, FNDC5 and BDNF proteins in the cerebral cortex and hippocampus (not in the local muscle) were significantly down-regulated in the model group (P<0.01, P<0.05). In comparison with the model group, the increase of Zea-Longa's score, Balance Beam score, percentage of cerebral infarct volume, and the decrease of expression levels of PGC-1α, FNDC5 and BDNF proteins in the ischemic peripheral cortex and that of BDNF in the hippocampus were reversed in the EA group (P<0.01, P<0.05). No significant changes were found in the expression levels of hippocampal PGC-1α and FNDC5 proteins in the hippocampus and those of PGC-1α, FNDC5 and BDNF proteins in the local muscle after EA intervention (P>0.05). CONCLUSION: EA can improve neurological and motor functions and reduce cerebral infarction volume in CI/RI rats, which may be related to its functions in activating PGC-1α/Irisin(FNDC5)/BDNF pathway in the cerebral cortex.

Strigolactone GR24 improves cadmium tolerance by regulating cadmium uptake, nitric oxide signaling and antioxidant metabolism in barley (Hordeum vulgare L.).

Cadmium (Cd) in the food chain poses a serious hazard to human health. Therefore, a greenhouse hydroponic experiment was conducted to examine the potential of exogenously strigolactone GR24 in lessening Cd toxicity and to investigate its physiological mechanisms in the two barley genotypes, W6nk2 (Cd-sensitive) and Zhenong8 (Cd-tolerant). Exogenous application of 1 µM GR24 (strigol analogue) reduced the suppression of growth caused by 10 µM Cd, lowered plant Cd contents, increased the contents of other nutrient elements, protected chlorophyll, sustained photosynthesis, and markedly reduced Cd-induced H2O2 and malondialdehyde accumulation in barley. Furthermore, exogenous GR24 markedly increased NO contents and nitric oxide synthase activity in the Cd-sensitive genotype, W6nk2, effectively alleviating the Cd-induced repression of the activities of superoxide dismutase and peroxidase, increasing reduced glutathione (GSH) and ascorbic acid (AsA) pools and activities of AsA-GSH cycle including ascorbate peroxidase, glutathione peroxidase, glutathione reductase, dehydroascorbate reductase and monodehydroascorbate reductase. The findings of the present study indicate that GR24 could be a candidate for Cd detoxification by decreasing Cd contents, balancing nutrient elements, and protecting barley plants from toxic oxidation via indirectly eliminating reactive oxygen species (ROS), consequently contributing to reducing the potential risk of Cd pollution.

Differences in Post-ischemic Motor Recovery and Angiogenesis of MCAO Rats Following Electroacupuncture at Different Acupoints.

BACKGROUND: Electroacupuncture (EA) can promote nerve and vascular regeneration, confer neuroprotection, inhibit apoptosis and inflammatory reactions, reduce oxidative stress injury, regulate neurochemicals and inhibit the formation of brain oedema in cerebral ischemic. However, the precise site of EA stimulation in the treatment of cerebral ischemic is unclear. OBJECTIVE: In the present study, we investigated the effect of EA at the acupoints of different meridians in motor function recovery and the involvement of Vascular Endothelial Growth Factor (VEGF), phosphorylated Protein Kinase B (P-Akt), phosphorylated endothelial nitric oxide synthase (p-eNOS) and Platelet Endothelial Cell Adhesion Molecule-1(CD31) were examined in the peri-infarction cortex of rats. METHODS: The Middle cerebral artery occlusion (MCAO) model or sham surgery was performed in a total of Ninety male Sprague-Dawley rats. Rats were randomly divided into five groups: a sham group, a middle cerebral artery occlusion (MCAO) group, a Yang meridian group, a Yin meridian group and a combined Yang and Yin meridian group. EA stimulus was given during the middle cerebral artery occlusion. The neurobehavioural function was measured using Modified Neurological Severity Scores (mNSS), the rotarod test and the ladder rung walking test, and the protein expression of VEGF, P-Akt, p-eNOS in the peri-infarction cortex was detected by Western blot. Immunofluorescence was used to measure the vascular density of the peri-infarction cortex. RESULTS: EA at different meridian acupoints has no effect on the infarction volume, while EA at Yin meridian acupoints significantly promoted neurobehavioural functional recovery, increased the vascular density and enhanced protein kinase B/Endothelial nitric oxide synthase (Akt/eNOS) phosphorylation and VEGF expression. CONCLUSION: In the early stage of stroke, EA at Yin meridian acupoints can improve neurobehavioural functional recovery and the mechanism of this effect may be related to the enhanced expression of VEGF, P-Akt and p-eNOS in the peri-infarction cortex of rats.

An enriched environment increases the expression of fibronectin type III domain-containing protein 5 and brain-derived neurotrophic factor in the cerebral cortex of the ischemic mouse brain.

Many studies have shown that fibronectin type III domain-containing protein 5 (FDNC5) and brain-derived neurotrophic factor (BDNF) play vital roles in plasticity after brain injury. An enriched environment refers to an environment that provides animals with multi-sensory stimulation and movement opportunities. An enriched environment has been shown to promote the regeneration of nerve cells, synapses, and blood vessels in the animal brain after cerebral ischemia; however, the exact mechanisms have not been clarified. This study aimed to determine whether an enriched environment could improve neurobehavioral functions after the experimental inducement of cerebral ischemia and whether neurobehavioral outcomes were associated with the expression of FDNC5 and BDNF. This study established ischemic mouse models using permanent middle cerebral artery occlusion (pMCAO) on the left side. On postoperative day 1, the mice were randomly assigned to either enriched environment or standard housing condition groups. Mice in the standard housing condition group were housed and fed under standard conditions. Mice in the enriched environment group were housed in a large cage, containing various toys, and fed with a standard diet. Sham-operated mice received the same procedure, but without artery occlusion, and were housed and fed under standard conditions. On postoperative days 7 and 14, a beam-walking test was used to assess coordination, balance, and spatial learning. On postoperative days 16-20, a Morris water maze test was used to assess spatial learning and memory. On postoperative day 15, the expression levels of FDNC5 and BDNF proteins in the ipsilateral cerebral cortex were analyzed by western blot assay. The results showed that compared with the standard housing condition group, the motor balance and coordination functions (based on beam-walking test scores 7 and 14 days after operation), spatial learning abilities (based on the spatial learning scores from the Morris water maze test 16-19 days after operation), and memory abilities (based on the memory scores of the Morris water maze test 20 days after operation) of the enriched environment group improved significantly. In addition, the expression levels of FDNC5 and BDNF proteins in the ipsilateral cerebral cortex increased in the enriched environment group compared with those in the standard housing condition group. Furthermore, the Pearson correlation coefficient showed that neurobehavioral functions were positively associated with the expression levels of FDNC5 and BDNF (r = 0.587 and r = 0.840, respectively). These findings suggest that an enriched environment upregulates FDNC5 protein expression in the ipsilateral cerebral cortex after cerebral ischemia, which then activates BDNF protein expression, improving neurological function. BDNF protein expression was positively correlated with improved neurological function. The experimental protocols were approved by the Institutional Animal Care and Use Committee of Fudan University, China (approval Nos. 20160858A232, 20160860A234) on February 24, 2016.

[Micro-ribonucleic acids participate in electroacupuncture intervention-induced improvement of ischemic stroke].

Ischemic stroke is a major cause of permanent disability and death in adults, and electroacupuncture (EA) intervention has a positive role in improving neurological function in patients with ischemic stroke through a series of complex processes. In the present paper, we make a review about the development of researches on the involvement of micro-ribonucleic acid (miRNA) in ischemic stroke from excitatory amino acid toxicity, oxidative stress, inflammatory response, apoptosis and necrosis, and particularly sum up outcomes of researches about the roles of miRNAs in EA-induced improvement of neurological function in experimental cerebral ischemia animals. EA treatment can 1) balance levels of miRNAs (such as mir-126 and mir-328, etc.) to promote angiogenesis of ischemic cerebral cortex tissue by regulating expression of vascular endothelial growth factor family genes and proteins; 2) promote nerve regeneration by up-regulating serum miR-124 and hippocampal miR-132 expression to possibly facilitate cerebral repair and reduce cognitive dysfunction respectively via related proteins; 3) reduce cerebral edema via modulating expression of some miRNAs to control expression of aquaporin, matrix metalloproteinases, etc. and 4) suppress inflammatory response via up-regulating expression of miRNAs to inhibit expression of NF-κB, TNF-α, etc. in the local cerebral tissue. As a result, the neurological function is bettered after EA intervention.

[Change in ghrelin level with the amelioration of glucose and lipid metabolic disorder in OLETF rats].

OBJECTIVE: To explore the exact role of ghrelin in glyco- and lipo-metabolism. METHODS: We compared the levels of ghrelin mRNA in gastric tissue, ghrelin in gastric tissue and plasma among LETO rats( non diabetes, n=10), OLETF rats( type 2 diabetes, n=10), OLETF/M rats( OLETF rats managed with Metformin at the dose of 100 mg/kg weight, n=10) and OLETF/F rats( OLETF rats managed with Fenofibrate at the dose of 20 mg/kg weight, n=10). The levels of ghrelin mRNA were tested by Northern blotting, and the ghrelin content in gastric tissue and plasma detected by RIA. RESULTS: At the age of 30 weeks, the ghrelin fasting plasma levels of OLETF rats were lower than those of LETO rats(37.49+/-6.42 vs 58.52+/-5.85, P<0.05). The fasting blood plasma ghrelin levels of OLETF/M groups showed an increased tendency, but the difference of the fasting blood plasma ghrelin levels to those of the untreated OLETF rats were not noticeable(49.65+/-6.76 vs 37.49+/-6.42,P>0.05). However, the fasting blood plasma ghrelin levels of OLETF/F group were more than those that of the untreated OLETF rats (62.02+/-7.35 vs 37.49+/- 6.42,P<0.05). The mRNA and polypeptide levels of ghrelin in stomach tissue had no marked difference in the 4 groups at the age of 17 weeks. At the age of 30 weeks, the difference of mRNA(1.18+/-0.06 vs 1.27+/-0.05,P<0.05) and polypeptide (114.77+/- 31.65 vs 152.87+/-18.24, P<0.05) between OLETF group and LETO group had been observed. Metformin didn't influence the mRNA and polypeptide levels of ghrelin in stomach tissue markedly. Howere, the polypeptide (161.75+/-23.61 vs 114.77+/-31.65, P<0.05) and mRNA(1.36+/-0.09 vs 1.18+/-0.06,P<0.05) levels of ghrelin in stomach tissue of OLETF/F group were markedly higher than those of stomach tissue ghrelin in the untreated OLETF group. CONCLUSION: Our data demonstrate that ghrelin may play a role in protective, compensative and negative-feedback regulation in the disorder of glucose and lipid metabolism.

[Effect of fenofibrate and metformin on lipotoxicity in OLETF rat kidney].

OBJECTIVE: To explore the possible molecular mechanism of abnormal renal lipid metabolism in OLETF rats, and the effects of fenofibrate or metformin on renal local lipid metabolism. METHODS: OLETF rats were randomly divided into three groups: one treated with fenofibrate, one with metformin and one without treatment. LETO rats were used as age-matched normal controls. The animals were sacrificed and the kidneys were obtained respectively at the age of 17 and 30 weeks. 24-hour urine albumin contents and TG contents in renal cortex were measured.The levels of SREBP-1 protein and mRNA in kidney were measured. FAS mRNA and ACC mRNA expressions were measured with RT-PCR. RESULTS: OLETF rats showed characters of obesity, hyperglycemia, hyperlipidemia. At the end of thirty weeks clinical characters of renal damage appeared in OLETF rats, OLETF rats showed higher levels of 24-hour urine albumin contents than those of LETO rats. 24-hour urine albumin contents were correlated positively with TG contents in kidney. The partial correlation coefficient was 0.870 (P = 0.011) by controlling for body weight, serum lipid, blood glucose and fasting insulin concentration. OLETF rats showed higher levels of SREBP-1 protein (43.2%, P<0.01), FAS mRNA (126.0%, P<0.01), ACC mRNA (72.3%, P<0.01) than those of group LETO. OLETF/F rats showed lower levels of SREBP-1 (15.7%, P = 0.061), FAS mRNA (36.8%, P<0.05), ACC mRNA (40.3%, P<0.05) than those of group OLETF. OLETF/M rats showed lower levels of SREBP-1 (19.3%, P<0.01), FAS mRNA (57.3%, P<0.01), ACC mRNA (22.5%, P<0.05) than those of group OLETF. There was no difference in SREBP-1 mRNA levels for each group. CONCLUSION: Renal damage occurred in OLETF rats at the prediabetes stage. 24-hour urine albumin contents were correlated positively with TG contents in kidney. SREBP-1 might aggravate fat deposition in kidney through some enzyme associated with fat synthesis, which was a possible pathogenesis of diabetic nephropathy. Treatment with fenofibrate or metformin ameliorated renal damage in OLETF rats through SREBP-1 and some enzyme regulated by it reduced fat deposit in kidney directly.

[Case-control study on manipulation following arthroscopic capsular release for the treatment of frozen shoulder].

OBJECTIVE: To compare therapeutic effects between the normal rehabilitation and combined with manipulative method after arthroscopic capsular release for the treatment of severe frozen shoulder, and to evaluate the application value of manipulationp. METHODS: From March 2007 to July 2010,arthroscopic capsular release was performed in 48 cases (48 shoulders, 23 left side, 25 right side). All the patients were divided into two groups: control group (11 males and 15 females) and manipulation group (9 males and 13 females). The patients in the control group were treated with conventional rehabilitation procedure, and the patients in the manipulation group were treated with additional manipulation procedure. From the 2nd day after operation, the manipulation was performed for 20 minutes every time, twice daily, and it continued for 10 days. All the cases were followed up and the scale of American Shoulder and Elbow Surgeons Standardized Assessment Form (ASES self-report section) and the range of motion (ROM) were recorded. RESULTS: The mean follow-up period was (12.54 +/- 5.78) months (ranging from 4 to 25 months). Both ASES scores and ROM in the manipulation group were better than those in the control group at the 1st month after operation, and the difference between the ASES scores and flexion of the shoulder were significant. However, there was no significan difference at the latest follow-up. CONCLUSION: Compared with the conventional rehabilitative procedure, manipulation following arthroscopic capsular release could promote the process of joint rehabilitation and help the patient back to normal life earlier, but there is no evidence of long term advantage.