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To provide markers to identify chromosomes in the genome of octopods, chromosomes of three octopus species were subjected to NOR/C-banding. In addition, we examined their genome size ( C value) to submit it to the Animal Genome Size Database. Silver staining revealed that the number of Ag-nucleoli was 2 ( Octopusminor (Sasaki, 1920)), 2 ( Amphioctopusfangsiao (d'Orbigny, 1839)) and 1 ( Cistopuschinensis Zheng et al., 2012), respectively, and the number of Ag-nucleoli visible was the same as that of Ag-NORs on metaphase plates in the same species. In all analyzed metaphases, Ag-NORs were mainly located terminally on the long arms of chromosomes 3 (3 rd ) of O.minor and on the short arms of chromosomes 4 (4 th ) of A.fangsiao , whereas only one of the chromosomes 23 (23 rd ) was found Ag-NORs of C.chinensis . C-bands were localized predominantly in the centromeric regions of chromosomes in the three species, while other conspicuous stable C-bands were observed in terminal regions, including the Ag-NORs. That means these three chromosome pairs (3 rd , 4 th and 23 rd ) could be considered species-specific cytogenetic markers. The mean C values of O.minor , A.fangsiao and C.chinensis were 7.81±0.39 pg (0.070 pg per unit length), 8.31±0.18 pg (0.068 pg per unit length) and 5.29±0.10 pg (0.038 pg per unit length), respectively, and results showed that C values of the three species were not proportional to the relative length of the chromosomes. These cytogenetic characteristics will provide more theoretical foundation for further researches on chromosome evolution in octopods.

Objective: To explore the specific mechanism of heat-reinforcing needling (HRN) for rheumatoid arthritis (RA) rabbits with cold syndrome based on metabolomics., Methods: A total of 40 healthy rabbits were randomly divided into a normal group, a model group, a reinforcing-reducing needling(RRN) group, a twirling-reinforcing needling (TRN) group and a HRN group ( n =8 rabbits/group). The RA model with cold syndrome was established with ovalbumin and freezing. Except the normal and model groups, RRN, TRN and HRN were given at "Zusanli" (ST 36) in the corresponding acupuncture groups for 30 min, once a day for 7 days. After the interventions, the pain threshold and the local skin temperature of each group were observed. The endogenous metabolites in the serum were analyzed by the gas chromatography mass spectrometry (GC-MS) technology combined with the pattern-recognition method., Results: The pain threshold and the local skin temperature in the model group were lower than those in the normal group ( P <0.05). The pain thresholds and the local skin temperatures in the three acupuncture groups were higher than those in the model group after interventions ( P <0.05), and those in the HRN group were obviously higher than those in the RRN and TRN groups ( P <0.05). The serum metabolites of leucine, valine, isoleucine, and arachidonic acid in the model group were increased compared with those in the normal group ( P <0.05), and the metabolites of α-ketoglutaric acid, citric acid, succinic acid, glucose, inositol, palmitic acid, stearic acid, lactose, d-ribose, and D-mannose were decreased ( P <0.05). The above-mentioned metabolites in the three acupuncture groups were significantly reversed relevant to the model group ( P <0.05), and the effect of HRN group was significantly superior to those of the RRN and TRN groups in regulating α-ketoglutaric acid, citric acid, succinic acid, glucose, inositol, d-ribose, and D-mannose ( P <0.05)., Conclusions: HRN for RA with cold syndrome is effective, which may be related to the specific regulation for the krebs cycle and glycometabolism.

Objective: To observe the influence of scalp-acupuncture on the expression of acid-sensing ion channels (ASICs) 1 a and 2 b of hippocampal CA 1 region in cerebral ischemia (CI) rats, so as to investigate its mechanism underlying improvement of ischemic stroke., Methods: Thirty-two male SD rats were randomly allocated to normal control, model, scalp-acupuncture and Amiloride group ( n =8 in each group). The model of focal CI was established by middle cerebral artery occlusion (MCAO). Scalp acupuncture stimulation was applied to bilateral Dingnieqianxiexian (MS 6) and Dingniehouxiexian (MS 7), once daily for 7 days. Rats of the Amiloride group were fed with Amiloride solution, twice a day for 7 days, and those of the normal control and model groups were grabbled and fixed in the same way with the acupuncture and Amiloride groups. The neurological deficit score was given according to Longa's method. The expression of hippocampal ASIC 1 a and ASIC 2 b was detected by immunohistochemistry, and the Ca 2+ concentration in the hippocampal tissue assayed using flowing cytometry., Results: After the intervention, the neurological deficit score of both the scalp-acupuncture and Amiloride groups were significantly decreased in comparison with pre-treatment ( P <0.01), and the effect of scalp-acupuncture was markedly superior to that of Amiloride in reducing neurological deficit score ( P <0.05). The expression of ASIC 1 a and ASIC 2 b in the hippocampal CA 1 region and hip-pocampal Ca 2+ concentration were significantly up-regulated in the model group compared with the normal control group ( P <0.01), and obviously down-regulated in both scalp-acupuncture and Amiloride groups ( P <0.01, P <0.05),without significant differences between the two treatment groups in the ASIC 1 a and ASIC 2 b expression and Ca 2+ concentration ( P >0.05)., Conclusions: Scalp-acupuncture stimulation can improve neurological function in CI rats, which may be related to its effects in suppressing the increased expression of hippocampal ASIC 1 a and ASIC 2 b proteins and in reducing calcium overload in hip-pocampal neurocytes.

Objective: To observe influences of heat-reinforcing needling (HRN) on scores of traditional Chinese medicine (TCM) symptoms and expression of plasma ATP synthase subunit O ( Atp 5 O ) mRNA and lysosomal V 1 subunit B 2 ( Atp 6 V 1 B 2 ) mRNA in patients with wind-cold-damp retention type rheumatoid arthritis (RA), so as to investigate its biological mechanisms in "heat production"., Methods: Sixty wind-cold-damp retention type RA patients were randomly allocated to HRN group ( n =30) and control group ( n =30). Guanyuan (CV 4), Qihai (CV 6), bilateral Zusanli (ST 36), and local acupoints near the knee-joint were selected for needling stimulation. Patients of the HRN group were treated by manipulating the acupuncture needle with HRN, and those of the control group treated by manipulating the needle with uniform reinforcing-reducing method. The treatments were performed once daily, 5 days a week, and two weeks altogether. The other 30 healthy volunteers were recruited as the normal control group. The TCM symptom scoring system (0-31 points, 11 items as the severities of pain, swelling and tenderness of the knee-joint) was used to evaluate the status of RA, and quantitative real-time PCR (RT-PCR) was used to detect the expression of plasma Atp 5 O mRNA and Atp 6 V 1 B 2 mRNA following removal of red blood cells., Results: After the treatment, the TCM scores of both the HRN and control groups were significantly decreased ( P <0.05), and that of the HRN group was significantly lower than that of the control group ( P <0.05). Before the treatment, the expression levels of plasma Atp 5 O mRNA and Atp 6 V 1 B 2 mRNA in RA patients were significantly lower than those of the normal group ( P <0.05), and after the treatment, the expression levels of plasma Atp 5 O mRNA and Atp 6 V 1 B 2 mRNA were significantly increased in both HRN and control groups compared to pre-treatment in the same one group ( P <0.05), and the up-regulated Atp 5 O mRNA and Atp 6 V 1 B 2 mRNA levels were remarkably higher in the control group than in the HRN group ( P <0.05)., Conclusions: Both heat-producing needling and uniform reinforcing-reducing needling can improve RA patients' clinical symptoms, which may be associated with their actions in up-regulating expressions of plasma Atp 5 O mRNA and Atp 6 V 1 B 2 mRNA.

Ischemic stroke is one of the commonly met diseases in clinical practice nowadays. Acupuncture therapy is widly used in the treatment of sequela of ischemic stroke in China and its mechanisms have been extensively studied in recen years. In the present paper, the authors focus on the development of studies on the mechanism of scalp acupuncture therapy in the treatment of ischemic stroke. Results indicate that scalp acupuncture intervention can 1) improve cerebral blood circulation to promote regional energy metabolism, 2) up-regulate expression of glial cell-line derived neurotrophic factor (GDNF), etc., possibly promoting proliferation and differentiation of neural stem cells in the focal cerebral cortex and hippocampus, 3) reduce contents of excitatory amino acid and increase level of gamma-aminobutyric acid (GABA) to lower neurogenic toxicity, and relieve cerebral injury, 4) ease cerebral vascular immunoinflammatory reactions, 5) regulate blood lipid metabolism to resist cerebral free radical damage, and 6) inhibit cerebral cortical apoptosis. However, these results only revealed very limited intrinsic mechanisms of scalp acupuncture in improving ischemic stroke. Further studies using comprehensive techniques of multi-disciplines as molecular biology, electrophysiology, etc. are definitely needed.

Objective: To observe the influence of scalp acupuncture on levels of high-sensitivity C-reactive protein (hs-CRP), TNF-α, IL-6, and IL-1β levels in patients with acute cerebral infarction (ACl), so as to investigate its mechanism underlying improvement of ACI., Methods: A total of 61 patients with ACI were randomly allocated to scalp acupuncture group (n = 31) and control (medication) group (n = 30). The patients of the control group were routinely treated by administration of Aspirin, Danhong injection, Cytidine Diphosphate for neurotrophy, blood pressure-control and blood-fat lowering medicines, etc., while those of the scalp acupuncture group were treated by routine treatment with the medicines mentioned above plus daily scalp acupuncture stimulation of bilateral Dingnieqianxiexian [MS 6, penetrative needling from Qianding (GV 21) to Xuanli (GB 6)] and Dingniehouxiexian [MS 7, from Baihui (GV 20) to Qubin (GB 7)]. The treatment was conducted once daily for 7 days. Serum hs-CRP, TNF-α, IL-6, and IL-1β contents were assayed by using enzyme linked immunosorbnent assay (ELISA). The therapeutic effects of scalp acupuncture were evaluated by using clinical neurological disfunction scale (NDS, 0-45 points for consciousness, gazing, facial palsy, speech, myodynamia, walking-ability)., Results: (1) Of the 30 and 31 cases in the control and scalp acupuncture groups, 5 (16.7%) and 8 (25.8%) were basically controlled, 9 (30.0%) and 16 (51.6%) experienced remarkable improvement in their symptoms, 12 (40.0%) and 6 (19.4%) were improved, 4 (13.3%) and 1(3.2%) failed, with the effective rates being 86.7% and 96.8%, respectively. The increased levels of serum hs-CRP, TNF-α, IL-6 and IL-1β in ACl patients were reversed on the 3rd and 7th day after scalp acupuncture treatment (P < 0.05, P < 0.01). (2) A positive correlation existed between the NDS score and the serum levels of hs-CRP (r = 0.497, P < 0.01). (3) NDS scores were obviously decreased in both groups on the 7th day after the treatment compared with their baseline data (P < 0.05, P < 0.01)., Conclusion: Scalp acupuncture treatment can improve the ACI patients' clinical symptoms, probably by reducing ACl induced inflammatory reactions.

It is important to real-timely monitor and control the temperature of cell physiological solution in patch clamp experiments, which can eliminate the uncertainty due to temperature and improve the measurement accuracy. This paper studies the influence of different ions at different concentrations in the physiological solution on precision of a temperature model by using near infrared spectroscopy and chemometrics method. Firstly, we prepared twelve sample solutions respectively with the solutes of CaCl2, KCl and NaCl at four kinds of concentrations, and collected the spectra of different solutions at the setting temperature range 20-40 degrees C, the range of the spectra is 9 615-5 714 cm(-1). Then we divided the spectra of each solution at different temperatures into two parts (a training set and a prediction set) by three methods. Interval partial least squares method was used to select an effective wavelength range and develop calibration models between the spectra in the selected range and temperature velues. The experimental results show that RMSEP of CaCl2 solution with 0.25 g x mL(-1) is maximum, the result of the three tests are 0.386 3, 0.303 7 and 0.337 2 degrees C, RMSEP of NaCl with 0.005 g x mL(-1) solution is minimum, the result of the three tests are 0.220 8, 0.155 3 and 0.145 2 degrees C. The experimental results indicate that Ca2+ has the greatest influence on the accuracy of the temperature model of the cell physiological solution, then K+, and Na+ has the least influence. And with the ionic concentration increasing, the model accuracy decreases. Therefore; when we build the temperature model of cell physiological solution, it is necessary to change the proportion of the three kinds of main ions in cell physiological solution reasonably in order to correct the effects of different ionic concentrations in physiological solution and improve the accuracy of temperature measurements by near infrared spectroscopy.

The biological effect on the organism generated by magnetic field is widely studied. The present study was aimed to observe the change of sodium channel under magnetic field in neurons. Cortical neurons of Kunming mice were isolated, subjected to 15 Hz, 1 mT pulse magnetic stimulation, and then the currents of neurons were recorded by whole-cell patch clamp. The results showed that, under magnetic stimulation, the activation process of Na(+) channel was delayed, and the inactivation process was accelerated. Given the classic three-layer model, the polarization diagram of cell membrane potential distribution under pulse magnetic field was simulated, and it was found that the membrane potential induced was associated with the frequency and intensity of magnetic field. Also the effect of magnetic field-induced current on action potential was simulated by Hodgkin-Huxley (H-H) model. The result showed that the generation of action potential was delayed, and frequency and the amplitudes were decreased when working current was between -1.32 μA and 0 μA. When the working current was higher than 0 μA, the generation frequency of action potential was increased, and the change of amplitudes was not obvious, and when the working current was lower than -1.32 μA, the time of rising edge and amplitudes of action potential were decreased drastically, and the action potential was unable to generate. These results suggest that the magnetic field simulation can affect the distribution frequency and amplitude of action potential of neuron via sodium channel mediation.

Aim: To investigate the effect of moxibustion on intestinal flora and release of interleukin-12 (IL-12) and tumor necrosis factor-α (TNF-α) from the colon in rat with ulcerative colitis (UC)., Methods: A rat model of UC was established by local stimulation of the intestine with supernatant from colonic contents harvested from human UC patients. A total of 40 male Sprague-Dawley rats were randomly divided into the following groups: normal (sham), model (UC), herb-partition moxibustion (HPM-treated), and positive control sulfasalazine (SA-treated). Rats treated with HPM received HPM at acupuncture points ST25 and RN6, once a day for 15 min, for a total of 8 d. Rats in the SA group were perfused with SA twice a day for 8 d. The colonic histopathology was observed by hematoxylin-eosin. The levels of intestinal flora, including Bifidobacterium, Lactobacillus, Escherichia coli (E. coli), and Bacteroides fragilis (B. fragilis), were tested by real-time quantitative polymerase chain reaction to detect bacterial 16S rRNA/DNA in order to determine DNA copy numbers of each specific species. Immunohistochemical assays were used to observe the expression of TNF-α and IL-12 in the rat colons., Results: HPM treatment inhibited immunopathology in colonic tissues of UC rats; the general morphological score and the immunopathological score were significantly decreased in the HPM and SA groups compared with the model group [3.5 (2.0-4.0), 3.0 (1.5-3.5) vs 6.0 (5.5-7.0), P < 0.05 for the general morphological score, and 3.00 (2.00-3.50), 3.00 (2.50-3.50) vs 5.00 (4.50-5.50), P < 0.01 for the immunopathological score]. As measured by DNA copy number, we found that Bifidobacterium and Lactobacillus, which are associated with a healthy colon, were significantly higher in the HPM and SA groups than in the model group (1.395 ± 1.339, 1.461 ± 1.152 vs 0.045 ± 0.036, P < 0.01 for Bifidobacterium, and 0.395 ± 0.325, 0.851 ± 0.651 vs 0.0015 ± 0.0014, P < 0.01 for Lactobacillus). On the other hand, E. coli and B. fragilis, which are associated with an inflamed colon, were significantly lower in the HPM and SA groups than in the model group (0.244 ± 0.107, 0.628 ± 0.257 vs 1.691 ± 0.683, P < 0.01 for E. coli, and 0.351 ± 0.181, 0.416 ± 0.329 vs 1.285 ± 1.039, P < 0.01 for B. fragilis). The expression of TNF-α and IL-12 was decreased after HPM and SA treatment as compared to UC model alone (4970.81 ± 959.78, 6635.45 ± 1135.16 vs 12333.81 ± 680.79, P < 0.01 for TNF-α, and 5528.75 ± 1245.72, 7477.38 ± 1259.16 vs 12550.29 ± 1973.30, P < 0.01 for IL-12)., Conclusion: HPM treatment can regulate intestinal flora and inhibit the expression of TNF-α and IL-12 in the colon tissues of UC rats, indicating that HPM can improve colonic immune response.

Objective: To investigate the application experience and clinical effects of hand-assisted laparoscopic radical resection for rectal cancer., Methods: The clinical data of 156 patients with rectal cancer treated with hand-assisted laparoscopic surgery between August 2009 and April 2011 were analyzed retrospectively., Results: The operative procedures of 156 patients were completed successfully and 1 case was converted to laparotomy (0.6%). The mean operation time was (125 ± 35) minutes; the mean intraoperative blood loss was (118 ± 60) ml; the mean time of bowel function recovery was (60 ± 8) hours; the median postoperative hospital stay was (9.5 ± 2.2) days. The mean number of lymph nodes dissection was (14 ± 5). Five patients (3.2%) had postoperative complications. All the patients were followed up. There had been no local recurrence or trocar site implantation metastasis., Conclusion: Hand-assisted laparoscopic surgery approach for the curative resection of rectal cancer is safe and effective and has broad prospects in clinical application.

Aim: To investigate the effects of moxibustion on down-regulation of the colonic epithelial cell apoptosis and repair of the tight junctions in rats with Crohn's disease (CD)., Methods: Sixty male Sprague-Dawley rats were randomly divided into a normal control (NC) group, a model control (MC) group, an herbs-partitioned moxibustion (HPM) group, a mild-warm moxibustion (MWM) group and a salicylazosulphapyridine (SASP) group, with 12 rats in each group. The CD model rats were treated with trinitrobenzene sulphonic acid to induce intestinal inflammation. The rats in the HPM and MWM groups were treated at the Tianshu (ST25) and Qihai (CV6) acupoints once daily for 14 d, and the SASP group was fed SASP twice daily for 14 d. No additional treatment was given to the MC and NC groups. The microstructure of the colonic epithelium was observed under a transmission electron microscope, the transepithelial resistance was measured using a short-circuit current, colonic epithelial cell apoptosis was determined by terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end labelling assay, and the expression of occludin, claudin-1 and zonula occludens-l (ZO-1) in the colonic epithelial junction was determined by Western blotting and immunofluorescence staining., Results: Compared with the MC group, the microstructure of the colonic epithelial barrier was significantly improved in rats treated with HPM, MWM or SASP, meanwhile, the current flow was reduced significantly, with values of 168.20 ± 6.14 vs 99.70 ± 3.13, 99.10 ± 4.28 and 120.30 ± 3.65 mA, respectively (P = 0.001). However, the HPM and MWM groups had higher current flow rates than the SASP group (99.70 ± 3.13, 99.10 ± 4.28 vs 120.30 ± 3.65 mA, P = 0.001). The number of the apoptotic colonic epithelial cells in HPM, MWM and SASP groups was largely reduced (61.5 ± 16.91 vs 15.5 ± 8.89, 14.8 ± 6.27 and 24.7 ± 9.68, respectively (P = 0.001); and the expression of occludin, claudin-1 and ZO-1 in the MWM and HPM groups was significantly enhanced (0.48 ± 0.10, 0.64 ± 0.09 vs 0.18 ± 0.05 for occludin, 0.12 ± 0.02, 0.17 ± 0.03 vs 0.05 ± 0.01 for claudin-1, and 0.08 ± 0.01, 0.11 ± 0.01 vs 0.02 ± 0.01 for ZO-1). And in SASP group, the expression of occludin and ZO-1 was also significantly increased (0.27 ± 0.04 vs 0.18 ± 0.05 for occludin and 0.05 ± 0.01 vs 0.02 ± 0.01 for ZO-1), but there was no significant difference for claudin-1. The HPM and MWM groups had higher expression of occludin, claudin-1 and ZO-1 than the SASP group., Conclusion: HPM and MWM treatment can down-regulate apoptosis of colonic epithelial cells, repair tight junctions and enhance colonic epithelial barrier function in rats with CD.

The coarse woody debris (CWD) loading capacity of main forest types and of different larch forest communities in Huzhong area of Great Xing'an Mountains was investigated, with its environmental gradient analyzed by Detrended Canonical Correspondence Analysis (DCCA). The CWD loading capacity varied significantly with forest type, being the highest (0.20 m3 x hm(-2)) in spruce forest (Picea koraiensis). Different larch forest (Larix gmelinii) communities had a CWD loading capacity from 0 to 0.28 m3 x hm(-2), with the highest in Pinus pumila-Larix gmelinii (0.28 m3 x hm(-2)) and the lowest in Sphagnum magellanicum-Ledum palustre-Larix gmelinii (0), but the differences were not significant. The CWD loading capacity across the study area was complicated, which might result from the complex interplay of affecting factors. The main factors affecting the spatial pattern of the CWD loading capacity were topographic factors (elevation and slope position) and stand structure (age, canopy cover, and others), and the interaction between topographic factors and stand structure expressed the spatial gradient of CWD loading capacity in Huzhong area of Great Xing' an Mountains.

Objective: Extrahepatic bile duct carcinoma is a rare but dismal malignacy. This study is conducted to show retrospective review and analysis of the correlation between the prognosis and different treatment modalities., Methods: The data of 84 such patients treated by different modalities from January, 1992 to July, 2000 were retrospectively reviewed and analyzed using SPSS 10.0 statistical package. The survivals were estimated by the Kaplan-Meier method and the difference among groups was tested by the log-rank test. The prognostic factors were determined by Cox multivariate analysis., Results: Of the 84 patients, 33 had complete resection, 19 palliative resection, 12 exploration alone, and the remaining 20 were treated by chemotherapy and/or radiotherapy. The mean follow-up time was 592 days. The overall 5-year survival rate was 13.1%. The 1-, 3- and 5-year survival rate following complete resection was 76.8%, 52.6% and 30.5% respectively, which was significantly higher than those of palliative surgery or chemotherapy/radiotherapy (P < 0.01). Multivariate analysis revealed that lymph node status (P = 0), histopathological grade (P = 0.001) and distant metastasis (P = 0.002) were significant high risk factors., Conclusion: The prognosis of extrahepatic bile duct carcinoma remains poor even after complete resection as shown to have a 5-year survival of 30.5%. More effective adjuvant therapy is needed. Extended resection may be helpful in improving the prognosis for carefully selected patients. Early diagnosis and early treatment is still the key to improve the long-term survival of extrahepatic bile duct carcinoma.