Objective To study the microsurgical and anatomic structures of brachial plexus roots and vertebral canal to find the best approaches for reimplantation of avulsed brachial plexus ventral roots into the spinal cord.Methods On nineteen cervicothoracic spine specimens,the brachial plexus nerves were exposed along to intervertebral foramen,and the spinal cord and brachial plexus roots were exposed by excising the vertebral arch and sectioning the spinal dura mater.The anatomy of brachial plexus roots and vertebrae,and the relative positions of spinal cord segments to vertebral discs were measured and observed.Results The relative positions of spinal cord segments to vertebral discs are:C5-7 spinal cord segments face C3,4,C4,5 and C5,6 vertebral discs;and C8 and T1 spinal cord segments face C6 and C7 vertebrae.Based on the anatomic finding,four approaches were found out: the lateroventral approach,the lateral approach by enlarging intervertebral foramen,the laterodorsal approach and the lateral and dorsal combined approach.Conclusions The brachial plexus ventral roots can be best reimplanted into the spinal cord by the lateroventral approach and the lateral approach.Although the laterodorsal approach and the lateral and dorsal combined approach are not the best,they are less difficuh and dangerous.

Objective To confirm whether the open-ended coaxial line is effective in detection of the differences in dielectric properties between colorectal cancer tissues and surrounding normal tissues and evaluation of the depth of tumor invasion. Methods The open-ended coaxial line system at frequencies ranging from 50 to 500 MHz in 98 freshly excised colorectal cancerous specimens obtained from the operating theatre of Zhujiang Hospital, was used to detect both the relative permittivity and conductivity on the serosal surface of the carcinoma nidus, the mucosa of the carcinoma nidus, and the mucosa of the surgical resection margin. Pathological examinations were conducted on each specimen after surgery. Results The values for relative permittivity and conductivity of the colorectal cancerous mucosa were significantly higher than those of the normal mucosa (P < 0.01). For the tumor which had invaded or penetrated the serosa (stage ≥ T3), the dielectric properties of both the cancerous serosa and mucosa were higher than the one restricted to muscularis propria or less intestine wall (stage < T3) over the measured frequency range, and there existed statistical differences at the common frequencies of 213 MHz and 426 MHz. Conclusion The open-ended coaxial line system may result in fast and effective diagnostic differentiation between cancerous and normal colorectal tissues as well as reasonable assessment of the tumor infiltration depth.

Objective To investigate the effect of moderate pain on attentional bias towards emotional pictures among healthy subjects.Methods Thirty-two healthy college students aged from 17 to 26 (21.8±2.2;16 males and 16 females) participated in this study.A tourniquet was tied to each subject's left upper arm 1 to 2cm above the cubits horizontal grain.Pain was inflicted by inflating the tourniquet,and the pressure was maintained at 26.66kPa.While tourniquet was inflated (with pain) or not (no pain),each subject was asked to finish a pictorial dot-probe task with three kinds of pictures-emotionally positive,negative and neutral.In experiment 1,subjects performed the dot-probe tasks with the contralateral hand while the tourniquet was tied on the left upper arm without inflation.In experiment 2 the tourniquet was inflated until the subject completed the dot-probe task (for about 10min).The reaction times (RTs) and the error rates (Ers) in the recognition task were recorded,and the intensity of the subject's pain and discomfort were measured using a verbal rating scale.Results The subjects reported moderate to severe pain with the tourniquet inflated.The RT and ER data were analyzed using two-way analysis of variance (ANOVA) which showed a significant difference between the average RTs of the males (482±73ms without pain and 466±82ms with pain) and those of the females (536±90ms without pain and 519±100 ms with pain).The average ER was significantly different between the pain (2.38±1.49)％ and no pain (1.09±0.82)％ conditions in both groups.Holn-Sidak multiple comparison testing showed significant differences in both groups' average ER between the negative picture (3.81±1.73)％ and the positive picture (1.66±0.97)％,and between the negative and neutral pictures (1.68±0.8) ％ in the pain condition.Mild attentional avoidance was observed with the positive [pain condition (-5.1±4.8) ms and no pain (-4.6±4)ms] and negative pictures [pain condition (-3.43±6) ms and no pain (-0.79±4.1)ms],but no significant difference was found between the pain and no pain conditions.Conclusion The error rate in a pictorial dot-probe task is influenced by pain,especially with negative pictures.

8 ?g/ml were 7 strains in prophylactic treatment group and 3 strains in non-fluconazole prophylactic treatment group respectively.The two groups had significant difference (x~2=8.75,P

Objective To establish a method for detecting HBV cccDNA in hepatocytes of chronic hepatitis B patients.Method 21 liver biopsies from the hepatic operation patients in the hospital of jiangsu province,concluding 19 HBV chronic infected patients (10 HBeAg positive patients and 9 HBeAg negative patients) and 4 uninfected patients,HBV DNA(+) serum of hepatitis B patients was thought as rcDNA.To use proteinase K to release HBV cccDNA and genomic DNA,then divide the cell lysis solution into two parts,one for detecting HBV cccDNA,the other for detecting the number of ?-Globin as internal control. Nucleic acid for detecting HBV cccDNA extracted by phenol-chloroform was digested by plasmid-safe ATP dependent DNase which was applied to digest the single strand DNA in rcDNA and ssDNA,then was quantitated by the primers spanning across the nick and SYBR Green Ⅰ dye.The specifity of PCR production was confirmed by the sequence analysis and rcDNA comparison.The significance of the difference of HBV cccDNA level between HBeAg(+) and HBeAg(-) group was analyzed by two group t test.Results The agarose gelelectrophoresis showed the molecular weight of the PCR production was about 350bp.The coincidence rate of PCR production and goal fragement was nearly 99% by sequence analysis.The result of PCR detection of rcDNA group was negative.The positive rate of HBV cccDNA of liver biopsies of HBeAg (+) patients detected by this method was 100%,the level of HBV cccDNA in the liver biopsies of HBeAg (+) patients was higher than HBeAb(+) patients.Conclusions The specificity of the method is proved by agarose electrophoresis,gene sequencing of the PCR product and rcDNA comparison.The quantitative method that use SYBR Green Ⅰ dye and ?-Globin as internal control is more specific,sensitive and economical,and more suitable for clinical purpose.

Objective To develop a nylon membrane chip for rapid and systematic detection of the diabetes-associated 45 mutant loci in mitochondrial DNA(mtDNA).Methods The mutant-and wild-type probes were designed for detection of 45 mutant loci in mtDNA with Primer Premier 5.0 and NCBI BLAST softwares and the 90 probes with 8 poly T were immobilized on the Hybond N~+ nylon membranes which were treated with 5?SSC Buffer by UV-crosslinking;Then asymmetric PCR was employed to obtain the target single strand DNA(ssDNA).The PCR products were labeled with biotin after purification.NBT/BCIP was used as substrate that yields a very intense purple signal followed by AP-avidin,and the signals were observed in 24 samples with known sequences to evaluate the chips,each sample was repeatedly measured three times.Results The specific target fragments of 45 loci can be amplified under the same condition with nine sets of primers.The annealing temperatures of the wild-type [(59.01?1.42)℃] and mutant-type [(59.34?1.29)℃ ] probes are so close(t=1.046,P =0.301)that hybridization can be performed at the same temperature.The spots on the membrane chip are distinct,regular and well-distributed.The results of positive-and negative-control are perfect.The signals of negative probes and the background are similar.The results of chip were nearly concordant with that of DNA sequences(?~2=113.132,Kappa value =0.888,P = 0.000)except for T16189C mutant.Conclusions We have successfully developed a nylon membrane chip for rapid and systematic detection of the diabetes-associated 44 mutant loci in mtDNA.It could be used for screening for diabetic patients and high-risk people.

Objective To investigate the relationship between the genetic predisposition to type 2 diabetes mellitus(T2 DM)and mitochondrial DNA base variants in elderly patients.Methods PCR restriction fragment length polymorphism(PCR-RFLP)analysis was used to screen base variants at position 3243 and 3426 of mitochondrial DNA in 186 elderly cases with T2 DM and 170 healthy controls,and DNA sequence was confirmed.Results No carrier of 3243 A→G variant or 3426 A→G variant was found in both groups,however there was 1 case with 3290 T→C variant in diabetic group.Conclusions No significantly association was found between mitochondrial DNA 3243 or 3426 base variants and the predisposition of type 2 diabetes mellitus in elderly patients.

Objective To summarize experiences of microsurgical treatment of dumbbell tumors of the high cervical spine. Methods A series of 12 patients with dumbbell tumors of the high cervical spine were treated by using microsurgical techniques through posterior approach or antero-lateral approach. Results Complete resection was achieved in 10 patients. Postoperative neurological symptoms improved greatly in all. Conclusion The key points of treatment in dumbbell tumors of the high cervical spine are to analyze the preoperative image carefully and have knowledge about anatomy of high cervical spine as well as the experience of microsurgical technique.

Background Doxycycline is a broad spectrum antibiotic,and it is frequently used in the treatment of ocular surface diseases.Objective The purpose of the present study was to investigate the effect of doxycycline on the inhibition of cell proliferation in bovine corneal myofibroblasts in vitro and assess its contribution to ocular surface repairing mechanism.Methods Six fresh bovine corneas were collected.The corneal stromal layer was isolated by two-step method of 1.0 g/L and 2.0 g/L collegenase-1.Isolated cells were plated at mantaryay culture flask in 10％ FBS of RPMI-1640.Vimentin and alpha-smooth muscle actin (α-SMA) organization were evaluated by immunocytochemistry,and the cells with influoresccence staining for vimentin and α-SMA were identified as the corneal myofibroblasts.Doxycycline at the concentrations of 10,20,40,60,80 mg/L was added to the medium,respectively,in different concentrations of doxycycline groups.Dexamethasone (120 mg/L)was used in the same way in the positive control group,and no drug was used in the negative control group.Cell proliferation was evaluated by MTT and the cell cycle was analyzed by BD FACScan flow cytometer assay 24 hours and 48 hours after addition of any drug.Results The cells grew well and showed the positive response for vimentin and α-SMA.MTT assay showed that the A570values of bovine corneal myofibroblasts were gradually declined with the increase of the concentration of doxycycline and lapse of active time,showing statistically significant difference (Fconcentration =1233.778,P＜0.001 ; Ftime =227.564,P ＜ 0.001).And the difference between the two factors was also statistically significant (Ftime*concentration =51.656,P＜0.001).Flow cytometry cell cycle analysis showed that 24 hours after 10,20,40,60,80 mg/L doxycycline treated,the perentage of of corneal myofibroblast cell in G0-G1 phase was 82.85％,84.36％,85.18％,87.12 ％ and 89.31％,showing significant increase in comparison with 63.89％ of the negative control group (all P＜0.05),and that of 40 mg/L doxycycline group was near the positive control group.Forty-eight hours after 10,20,40,60,80 mg/L doxycycline treated,the perentage of of corneal myofibroblast cell in G0-G1 phase was 82.78％,86.15％,88.23％,89.57％,93.00％,with significant increase in comparison with 70.17％ of the negative control group (all P ＜ 0.01),and that of 40 mg/L doxycycline group was near the positive control group.Conclusions The growth of the bovine corneal myofibroblasts is inhibited by doxycycline in time-and dosedependent manner in the range from 10 mg/L to 80 mg/L,and 40 mg/L of doxycycline has an obviously inhibitory action as 120 mg/L dexamethasone.

BACKGROUNDThe development and maintenance of spiral ganglion cells (SGCs) appear to be supported by neurotrophins. Removal of this support leads to their gradual degeneration. Intracochlear infusion with neurotrophins can provide trophic support to SGCs in animal deafness models if given shortly after deafening. However, it is not known whether delayed intervention will provide similar protection, which might be clinically relevant. The present research was conducted to determine the effects of brain-derived neurotrophic factor (BDNF) administration on the capacity of the peripheral processes to resprout.

METHODSThe left cochlea of 20 profoundly deafened rats, which were divided into 2 groups equally, was implanted with an electrode and drug-delivery system 30 days after deafening. Either BDNF or artificial perilymph (AP) was delivered continuously for 28 days. Electrically evoked auditory brainstem responses (EABRs) were recorded during the period. SGC body and peripheral process density were measured.

RESULTSThe EABR thresholds of AP increase continually. Those of BDNF increase slowly at the beginning then decrease, and were significantly less than those of the AP group from day 14 to 28 (P < 0.01). In terms of SGC and peripheral process density, the difference between the treated and control ears of BDNF group was clearly significant (P < 0.01), but not in AP group (P > 0.05). Analysis of the left cochlea between the two groups demonstrated that SGC/peripheral process density of the BDNF group was significantly greater than that of the AP group. Finally, a functional formula was developed relating the last EABR threshold and SGC density and process density, which was as follows: T = 466.184 - 2.71 (F.B.L).

CONCLUSIONSUnder the conditions of delayed intervention following 30 days after deafening in rats, it can be concluded that BDNF enhances SGC bodies and peripheral processes survival after differentiation and so improves auditory sensitivity. SGC peripheral processes influence the auditory sensitivity.

Animals ; Brain-Derived Neurotrophic Factor ; administration & dosage ; Cell Survival ; drug effects ; Evoked Potentials, Auditory, Brain Stem ; drug effects ; Male ; Rats ; Rats, Sprague-Dawley ; Spiral Ganglion ; cytology ; drug effects