Objective To investigate the effect of stellate ganglion block (SGB) on substance P of spinal cord in rabbits following formalin stimulation and the possible mechanism. Methods Rabbits of both sexes weighting 2.5-3.0 kg were anesthetized with 20% urethane 1 g?kg-1 . Spontanous breathing was maintained. Right stellate ganglion was exposed aseptically. An epidural catheter was fixed with one end placed closed to stellate ganglion and the other end outside the neck through a hole on the skin for administration of drugs. One week later, nineteen healthy rabbits were randomly divided into three groups: sham operation group (group A, n = 5), SGB group (group B, n=7) and control group (group C, n = 7) . In group B and group C, 3 % formalin 0.2 ml was injected subcutaneously into plantar region of the right paw. Pain response was observed. 1h after formalin injected, 0.25% bupivacaine 0.5 ml was injected through the catheter in group B while normal saline 0.5 ml was injected in group C. The effect of SGB was confirmed by ptosis and miosis. 1h after using of bupivacaine or normal saline, rabbits were deeply anesthetized and chest was opened, 1 000 ml of normal saline was infused via left ventricle then followed by 2000-2500 ml of 4% paraformaldehyde in 0.1mol/L phosphate buffer pH7.4 infusion for fixation of tissue, 60 min later spinal cord of cervical 6-8(C6-8) and thoracic 6-8(T6-8 ) were removed. In group A the spinal cord were extracted similar to the other groups. The content of SP in spinal cord use immumohistochemistry technique of strept avidin biotin enzyme complex (SABC) to display. Results Immunoreaction of SP were distributed mainly in lamina Ⅰ and Ⅱ of dorsal horn of spinal cord. Compared with group A, the optical density(OD) of SP in cervical slices were decreased significantly in group B and group C(P 0.05 ). Conclusions The content of SP in spinal cord decreased significantly after subcutaneous injection of formalin in rabbits. SGB can reverse the phenomena partly and increase the content of SP of cervical spinal cord in rabbits following formalin stimulation. This may be the possible mechanism of analgesia provided by SGB.

Objective To investigate the effect of stellate ganglion block(SGB) on the plasma concentration of noradrenaline(NE) in rabbits suffering from acute pain and the possible mechanism.Methods Fourteen healthy rabbits of both sexes weighing 2.5 2.8 kg were anesthetized with 20% urethane 1 g?kg -1 . Spontanous breathing was maintained. Right stellate ganglion was exposed aseptically. An epidural catheter was fixed with one end placed close to stellate ganglion and the other end outside the neck through a hole on the skin for administration of drugs. One week later 3% formalin 0.2ml was injected subcutaneously into plantar region of the right paw. Pain response was observed . 60 min after formalin injection 0.25% bupivacaine 0.5ml was injected through catheter (bupivacaine group n=7) while in control group (n=7) normal saline 0.5ml was injected. The effect of SGB was confirmed by ptosis and miosis. Blood samples were taken from edge vein of the ear 10 min before (T 0) and 10(T 1), 30(T 2), 50min(T 3) after subcutaneous injection of formalin and 10min(T 4), 30min(T 5), 50min(T 6) after bupivacaine or normal saline injection for determination of plasma NE concentration by radioimmunoassay.Results Plasma NE concentration increased significantly after subcutaneous formalin injection and peaked at T 1,then decreased slightly at T 2 and T 3. In group B, plasma NE concentration decreased significantly after bupivacaine injection, while in control group there was no significantly change in plasma NE concentration after normal saline injection.Conclusions SGB reduces the increased plasma NE concentration in rabbits suffering from acute pain. This may be the possible mechanism of analgesia provided by SGB.

Objective: To analyze the influence of semen processing methods on the proportion of the aneuploid sperm by detecting the sperm's chromosome X, Y, 18 using fluorescence in situ hybridization. Methods: Ten patients with mild ohgoasthenosperia, who were received ICSI treatment, were included in this study. Five semen samples of the patients were randomly selected to detect using Swim-up method (A group) and 5 using sperm-grad double-density centrifugation method (B group).Another 5 patients with mild oligo-asthenosperia were as control (C group). The CEPX / Y and CEP18 probe was used to detect the sperm of these 15 patients by fluorescence in situ hybridization. The proportion of aneuploid sperm was compared in three groups. Results: The sex chromosome aneuploid rates were (4.21±2.49)%, (3.24～1.49) % and (2.62±0.89) % in control, A and B groups. The rates of aneuploid chromosome 18 were (3.00±1.22)%, (2.00～1.22)% and (2.00±1.22)% in control, A and B groups. There were no significant differences in three groups (P＞0.05). Conclusion: The results showed that the methods of Swim-up and Sperm-Grad double-density gradient centrifugation could select sperms in motility potential and teratospermia,but not in normal chromosome sperms.

BACKGROUND: Stellate ganglion block has many functions by improving brain circulation, modulating immunity, reducing plasmic catecholamine content, interleukin-6 is one of the most sensitive and important predictors and mediators for acute organic stress response, playing neuroprotective and neurotoxic double roles in brain ischemic injury.OBJECTIVE: To observe the effect of stellate ganglion block in rat brain during ischemic-reperfusion period on the changes of serum interleukin-6,in order to probe the role of stellate ganglion block in brain ischemicreperfusional injury.DESIGN: Randomized controlled experimentation.SETTING: Anaesthesia Department of Taihe Hospital Mfiliated to Yunyang Medical College, and Anaesthesia Department of Renmin Hospital Affiliated to Yunyang Medical College.MATERIALS: This experiment on animals was carried out at the Experimental Center of Taihe Hospital Affiliated to Yunyang Medical College at March 2003, interleukin-6 reagent kit and determination was provided and conducted by the immunity research institute of Chinese People's Liberation Army General Hospital. Totally 28 big-ear healthy rabbits in which male or female was not limited were selected and randomly divided into stellate ganglion group, saline comparison group, blank comparison group and sham operation group with 7 rabbits in each group.METHODS: A pipe was set approximate to the stellate ganglion of all animals by operative method, six-vessels block method was used to simulate whole brain ischemic-reperfusion model, in stellate ganglion block group,artery clamp was lossen for reperfusiion at 15 minutes after ischemia, simultaneously 2.5 g/L bupivacaine was continuously pumped into left side of stellate ganglion for nerve block, which replaced by physical saline and nothing in respectively physical saline comparison group and blank comparison group, while rabbits in sham operation group were only subjected to surgery without artery clamp. RIA was used to determine serum interleukin-6 content at before ischemia, reperfusion of 10 minutes, 4 hours, 10hours, when 20 hours and 30 hours individually.MAIN OUTCOME MEASURES: Serum interleukin-6 content in each group at various post-reperfusional time points.RESULTS: Totally 28 big-ears white rabbits were enrolled in this experiment and all data was entered the result analysis. Interleukin-6 content was on an increasing tendency in all groups, while was higher in stellate ganglion block group than in sham operation group only at reperfusion 30time point, the difference has significance [(321±52) and (299±45) ng/L,P ＜ 0.05]; Comparing to pre-ischemic group, interleukin-6 in physical saline group began increase remarkably from onset of reperfusion 4 hours[(365±46) ng/L], but began obviously increase at reperfusion 10 hours in blank comparison group [(368±31) ng/L, P ＜ 0.05]. The difference of interleukin-6 among stellate ganglion block group, sham operation group,physical saline group and blank comparison group does not have statistical significance (P＞0.05); the level of interleukin-6 in physical saline group and blank comparison group were higher than in sham at all reperfusion 4-30 hours, and even higher than in stellate ganglion group after reperfusion 10 hours, the difference has significant meaning (P ＜ 0.05). Moreover the increase of interleukin-6 content in stellate ganglion block group was remarkably lower than physical saline comparison group and blank comparison group (P ＜ 0.05).CONCLUSION: Stellate ganglion block may obviously reduce serum interleukin-6 level in rabbit brain during ischemic-reperfusion period, implying stellate ganglion block has a certain protective and curative function on the whole brain ischemic-reperfusion damage, and considered as a promising way in the treatment of brain ischemic-reperfusion damage.

Objective To investigate the effects of adenovirus containing human beta-nerve growth factor (Ad-hNGFβ) gene on substance P (SP) and calcitonin gene-related peptide (CGRP) content of the spinal cord in a rat model of neuropathic pain by chronic constrictive injury (CCI). Methods Forty-eight male SD rats weighing 200-250 g were randomly divided into 3 groups (n=16 each) : group Ⅰ sham operation; group Ⅱ CCI and group Ⅲ CCI + Ad-hNGFβ gene IT. The animals were anesthetized with intraperitoneal choral hydrate 300-350 mg/kg. The right sciatic nerve was exposed and 4 ligatures were placed on the sciatic nerve at 1-2 nun intervals as described by Bennet and Xie[5]. In sham operation group, right sciatic nerve was exposed but not ligated. In group Ⅰ and Ⅱ artificial cerebrnspinal fluid was injected IT instead of Ad-hNGFβ gene. The behavior score and the paw-withdrawal latency (PWL) to radiant heat and mechanical stimulus were measured one day before operation and every 4 days within the 28 days after gene transfection. Four animals were killed at 4, 7, 14 and 28 day after IT gene transfection in each group and lumbar segment (L4-6 ) of the spinal cord was removed for determination of SP and CGRP content by immunohistochemistry. Results The behavior scores were significantly higher and PWL to radiant heat and mechanical stimulus were significantly lower in group Ⅱ and Ⅲ than in group Ⅰ. There was no significant difference in the behavior score and PWL to mechanical stimulus between group Ⅱ and Ⅲ while the PWL to radiant heat was significantly higher in group Ⅲ than in group Ⅱ. After operation SP and CGRP content were significantly higher in group Ⅱ and group Ⅲ than in group Ⅰ , and significangly lower in group Ⅲ than in group Ⅱ 7-28 days after operation. Conclusion The recomhinant Ad-hNGFβ gene transfection can attenuate heat hyperalgesia by reducing SP and CGRP content of the spinal cord in a rat model of neuropathic pain induced by CCI.

This study was aimed to observe the influence of β-sitosterol (BSS) on estrogen receptor (ER) positive the human breast cancer cell line T47D and to study its mechanisms. ER antagonist ICI182 780 was employed to observe the influence on the proliferation. Proliferations of T47D cells influenced by different concentrations of BSS were analyzed by MTT assay. Cell cycle analyses were examined by flow cytometry. The protein expression of cyclin D1 was measured by western blot analysis and cyclin D1 mRNA was quantified by real-time PCR assay. The results showed that BSS in high dose exhibited significant inhibitory effects that were partly antagonized by ICI182 780 and decreased the proliferative index on T47D cells. However, BSS in low dose obviously promoted the proliferation that was completely inhibited by ICI182 780 and increased the proliferative index on T47D cells. The mRNA and protein levels of cyclin D1 were increased in low-dose BSS. The effect was blocked by ICI182 780. It was concluded that BSS in low concentration had phytoestrogenic effect by up-regulating the expression of cyclin D1 via ER pathway.

Objective To compare the dosimetric parameters of volumetric modulated arc therapy(VMAT),fixed field intensity modulated radiation therapy(IMRT)and three-dimensional conformal radiotherapy(3D-CRT)in the radiotherapy for the patients with locally recurrent nasopharyngeal carcinoma, and to analyze their characteristics. Methods Twelve patients with locally recurrent nasopharyngeal carcinoma were treated with VMAT, IMRT and 3D-CRT plan designed by Pinnacle 9.2 and Preciseplan 2.03 treatment planning system.The dosimetric parameters of targeted volumes and organs at risk were compared between three groups. Results The conformation indexes (CI)of VMAT and IMRT plans were similar,and they were both better than 3D-CRT plan,the difference was significant(P<0.05).The homogeneity index(HI)in three groups were similar,there were no statistically significant differences between them(P>0.05).The monitor units(MU)and beam time in 3D-CRT group were better than those in other two groups,and VMRT group was better than IMRT group,the statistical differences were observed between three groups (P<0.05 ).There were no statistical differences of organs at risk such as brainstem and lens between three groups(P>0.05).The doses of the spinal cord,optic nerve,optic chiasm and temporal lobe of brain in VMAT and IMRT groups were better than those in 3D-CRT group,there were statistical differences between them(P<0.05),and the data in VMAT and IMRT groups were similar,and there were no statistical differences(P>0.05).Conclusion There are differences of the targeted dose distribution between the three kinds of radiation technology, while VMAT and IMRT plans can cover the targeted areas and reduce the received doses of organs at risk.The CI,MU and beam time of VMAT plan are better than those of IMRT plan. 3D-CRT plan only has advantage in the MU and beam time.

Objective To investigate the distribution and antimicrobial resistance profile of the common pathogens isolated during the period from 2009 to 2015.Methods All the bacterial strains isolated from pediatric inpatients in Beijing Children's Hospital during the period from 2009 to 2015 were analyzed. Antimicrobial susceptibility was determined by disk diffusion method and Phoenix 100 Automated Microbiology System. Results were analyzed according to the guidelines of CLSI (2014) using WHONET 5.6 software.Results The total strains were 26630. The most common gram-positive isolates were Streptococcus pneumoniae,Staphylococcusaureusand coagulase-negative Staphylococcus (CNS), while the most frequently isolated gram-negative microorganisms were Klebsiella spp.,Pseudomonas aeruginosa and Escherichia coli. The prevalence of S. pneumoniae was up to 25.7 % (4101/15973) in all respiratory tract specimens. About 50.2 % of the S. pneumoniae isolates were not susceptible to penicillin. The prevalence of methicillin-resistant strains was 20.6 % in S. aureus (MRSA) and 87.8 % in coagulase negative Staphylococcus (MRCNS) on average. The prevalence of MRSA increased from 11.1 % in 2009 to 29.8 % in 2015. No S. pneumoniae or staphylococcal strains were found resistant to vancomycin or linezolid. The Enterococcus strains were still highly susceptible to vancomycin and linezolid. Overall 0.3 % of the Enterococcus faecium isolates were resistant to vancomycin. The extended-spectrum beta-lactamases (ESBLs) producing strains accounted for 71.4 % -78.1 % of E. coli and 65.1 % - 76.9 % of K. pneumoniae isolates. The carbapenem-resistant E. coli and K. pneumoniae were reported for the first time in 2010, but in 2014, the strains resistant to carbapenems had increased to more than 7 % in E. coli, and higher than 20 % in K. pneumoniae. In 2015, up to 27.7 % and 25.7 % of P. aeruginosa isolates were resistant to imipenem and meropenem, respectively, and 59.9 % of the A. baumannii isolates were resistant to imipenem and meropenem. Beta-lactamase was positive in 46.3 % of the H. influenzae isolates. Conclusions MRSA and the carbapenem-resistant strains of E. coli,K. pneumoniae and A. baumannii are still on the rise in pediatric inpatients, which poses a serious threat to clinical practice and implies the importance of strengthening infection control.

Objective: To observe the change levels of nuclear factor-kappa B (NF-κB) p65 protein in cytoplasm and nuclear, phosphorylation of inhibitor of kappa B (p-IκB) protein and cytochrome C (Cyt-c) , cleaved cysteinyl aspartate specific proteinase-3 (Cleaved caspase-3) , B-cell lymphoma/leukemia-2 (Bcl-2) in cytoplasm in the process of N, N-dimethylformamide (DMF) -induced apoptosis in H9c2 cardiomyocytes, and explore the tentative mechanism of apoptosis. Methods: H9c2 cardiomyocytes were exposed to 200 mmol/L DMF. Western blotting was used to detect the protein expression levels of p65 in cytoplasm and nuclear, p-IκB after exposure for 0, 2, 4, 6, 8, 12 h, and the protein expression levels of Cyt-c, Cleaved caspase-3, Bcl-2 in cytoplasm after exposure for 0, 2, 4, 8, 12, 24 h. Immunofluorescencecytochemistry (IFC) was used to observe the location of Cyt-c after 200 mmol/L DMF exposure for different times. Results: The levels of p65 in cytoplasm and nuclear and p-IκB among groups were statistically significant (F were 7.79, 33.11, 90.25, respectively, all P<0.01) . Compared with the control group, the levels of p65 in cytoplasm of 2, 4, 6 h group were significantly decreased (all P<0.01) ; the levels of p65 in nuclear of 2, 4, 6, 8 h were significantly increased (all P<0.01) ; the levels of p-IκB of 2, 4, 6 h group were significantly increased (all P<0.01) . The levels of Cyt-c, Cleaved caspase-3 and Bcl-2 among groups were statistically significant (F were 51.42, 503.68, 73.37, respectively, all P<0.01) . Compared with the control group, the levels of Cyt-c of 8, 12 h group were significantly increased (both P<0.01) ; the levels of Cleaved caspase-3 of 2, 4, 8, 12, 24 h were significantly increased (all P<0.01) ; the levels of Bcl-2 of 2, 4, 8, 12, 24 h group were significantly decreased (all P<0.01) . IFC showed that Cyt-c was released from the mitochondria to the cytoplasm gradually as the extension of the exposure time. Conclusion NF-κB signaling pathway and mitochondrial pathway are involved in the mechanism of DMF-induced apoptosis in H9c2 cardiomyocytes.

The rapid and accurate authentication of traditional Chinese medicines(TCMs)has always been a key scientific and technical problem in the field of pharmaceutical analysis.Herein,a novel heating online extraction electrospray ionization mass spectrometry(H-oEESI-MS)was developed for the rapid and direct analysis of extremely complex substances without the requirement for any sample pretreatment or pre-separation steps.The overall molecular profile and fragment structure features of various herbal medicines could be completely captured within 10-15 s,with minimal sample(＜0.5 mg)and solvent consumption(＜20 μL for one sample).Furthermore,a rapid differentiation and authentication strategy for TCMs based on H-oEESI-MS was proposed,including metabolic profile characterization,characteristic marker screening and identification,and multivariate statistical analysis model validation.In an analysis of 52 batches of seven types of Aconitum medicinal materials,20 and 21 key compounds were screened out as the characteristic markers of raw and processed Aconitum herbal medicines,respectively,and the possible structures of all the characteristic markers were comprehensively identified based on Com-pound Discoverer databases.Finally,multivariate statistical analysis showed that all the different types of herbal medicines were well differentiated and identified(R2X＞0.87,R2Y＞0.91,and Q2＞0.72),which further verified the feasibility and reliability of this comprehensive strategy for the rapid authentication of different TCMs based on H-oEESI-MS.In summary,this rapid authentication strategy realized the ultra-high-throughput,low-cost,and standardized detection of various complex TCMs for the first time,thereby demonstrating wide applicability and value for the development of quality standards for TCMs.