Objective To evaluate the normalized correction effect of the ROX reference fluorescence on the real-time quantitative RT-PCR assessment for the Nrf2 mRNA.Methods The Taq-man probe was utilized in the real-time quantitative RT-PCR method to assess the Nrf2 mRNA; The reaction specificity of the Nrf2 PCR products obtained from the rat cDNA was assessed using gel electrophoresis and sequencing identification.The PCR calibrators with different concentrations varied at 10 gradients(from 2.0 × 109 to 2.0 × 100) were prepared for real-time quantitative RT-PCR assessment,and then the influence of the ROX reference fluorescence correction on the linear range of the PCR standard curve was investigated experimentally.The recombinant plasmids with different concentration levels(e.g,high,medium or low concentration) were assessed by the real-time RT-PCR and 20 replicate measurements were performed for all the samples in both intra- and inter-assays.The repeatability differences of both the inter- and intra-assays were analyzed systemically with/without the ROX reference fluorescence correction.Results The primers and probes required for Nrf2 mRNA real-time quantitative RT-PCR assessments were constructed,and the reaction system and conditions were also established.The length of the amplified fragment agreed well with the expected length(122 bp),and the sequencing analysis showed the amplified fragment was verified by the sequencing identification.After applying the ROX reference fluorescence correction,the linear range ofthe PCR standard curve was measured as 2.0 × 109 -2.0 × 100 copies/μl(R2 ＞ 0.99),which was 100times wider than that measured without ROX reference fluorescence correction(2.0 × 109 - 2.0 × 102copies/μl,R2 ＞ 0.99).Without the ROX reference fluorescence correction,the intra-assay CVs of the Ct value were measured as 4.1%,2.7% and 2.1% for the recombinant plasmids with high-,medium-,and low-level concentration,respectively.The inter-assay CVs of the Ct value were 4.3% 、3.0% 、2.4%.By contrast,with the ROX reference fluorescence correction,the intra-assay CVs of the Ct value became 0.7%,0.5%,0.4% and the inter-assay CVs of the Ct value turned to 1.0%,0.8% and 0.7%.The discrete degree of the Ct value with the correction was lower than that without the correction.The intra- and interassay Ct values of the high- and medium-level samples exhibited statistic significance for the measurements with/without the correction.In the high-level group,the t values of the intra- and inter-assays were 4.843 and 2.566,with P＜0.05.In the medium-level group,the t values of the intra- and inter-assays were 4.293 and 4.423,with P＜0.05.However,no statistic significance was observed the low-level group(the t values of the intra- and inter-assays were 0.753 and 1.279,with P ＞.0.05).Conclusions The utilization of the ROX reference fluorescence in the real-time quantitative PCR assessment for Nrf2 mRNA could widen the linear range of the standard PCR calibration curve and improve the repeatability of the assessment,which should be quite helpful for accurately assessing the expression level of the Nrf2 mRNA.

Objective To explore the traditional Chinese and western medication principle of treating epilepsy based on text mining, and provide a reference for clinical use. Methods Based on the literature data of treatment of epilepsy collected in CBM database, we explored the medication principle of both traditional Chinese medicine and western medicine for treating epilepsy by frequency statistical data based on sensitive keywords hierarchical algorithm, retro-read and manually noise reduction. These laws displayed by the frequency of one-dimensional and two-dimensional network diagram. Results Commonly used drugs in western medicine treatment for epilepsy are phenobarbital, phenytoin, valproate, carbamazepine and diazepam. Traditional Chinese medicine commonly used are Angongniuhuang Pill, Qingkailing Injection, Wumei Pill, Tongxinluo Capsule, Compound Danshen Dripping Pill, Xuefuzhuyu Capsule, etc. When traditional Chinese medicine and western medicine are combined used, the most commonly used traditional Chinese medicine are Angongniuhuang Pill, Qingkailing Injection and Compound Danshen Tablet, western medicine are phenobarbital, valproate, carbamazepine and diazepam. Common syndromes are yin deficiency of liver and kidney, deficiency of kidney yin, heart tangled by sputum, deficiency of yin, heart disturbance by sputum and fire. Conclusion Text mining technology can be used for summary of medication principle of treating epilepsy and of epilepsy syndrome, thus provide useful exploration and reference for clinical diagnosis and treatment.

OBJECTIVE To observe the effect of vacuum-assisted closure in clinical persistent infection patients.METHODS Thirty two sufferers infected in soft tissue,bones and joints were treated by general methods but without durable concrescence.It lasted averagely 138 days.In our department it treated by vacuum-assisted closure in 23 days.Thereby a comparison of self-control came into being.RESULTS Thirty two cases were cured after following at least one year except for one case with recrudescence.CONCLUSIONS Vaccum-assisted closure is the most effective surgery therapy measurement and is a kind of unidirectional drainage.If we can use it felicitously then we can treat infection in orthopedics′ effectively.

Objective To investigate the expression of 5-lipoxygenase activating protein gene (ALOX5AP) polymorphism in the patients with cerebral infarction,and explore its relationship with cerebral infarction susceptibility.Methods Patients with cerebral infarction and healthy volunteers were selected for this study,whose venous blood was extracted and detected with polymerase chain reaction/restriction fragment length polymorphism (PCR-RFLP).Haplotype A (SG13S114T,SG13S89G,SG13S32A,SG13S25G),haplotype B (SG13S377A,SG13S114A,SG13S41A,SG13S35G),and their nucleotide polymorphism loci were observed.Results Single nucleotide polymorphism (SNP)-SG13S114,SNP-SG13S32 and HapA carrying rate were significantly different between patients with cerebral infarction and healthy volunteers (P ＜0.05).SNP-SG13S114 and SNP-SG13S32 were independent risk factors of cerebral infarction (OR ＞ 1.0,P ＜ 0.05).Conclusions The morbidity of cerebral infarction in Wenling City was influenced by SNP-SG13S114,SNP-SG13S32,and HapA carrying rate.

Objective To observe the effect of electric acupuncture on the infarct volume and amount of cerebral cortex and spinal neuron at different times of cerebral ischemia/reperfusion(I/R)in stroke prone renovascular hypertensive rats(RHRSP)with middle cerebral artery occlusion(MCAO),and investigate the possible mechanisms of electric acupuncture on remote damage in ischemic stroke. Methods 480 male SPF Sprague-Dawley(SD)rats were duplicated to form the RHRSP models by clamping both kidneys. 370 successful ones were selected by taking the tail artery blood pressure,and divided into hypertension group and sham operation group(each n=60)by random number table method. The MCAO models were created by stringing middle cerebral artery in the remaining RHRSP. The nerve function defect score(NDS)was graded by Longa 5 point method after the rats waked up from anesthesia,then the ones scored 1-3 were enrolled. Totally,there were 190 rats with MCAO successfully created from which 10 were randomly selected to determine the infarct size by 2,3,5-triphenyl four azole nitrogen chloride(TTC)staining. The remaining 180 MCAO rats were randomly divided into model group,electric acupuncture group and fake acupuncture group(each n=60). The sham operated group only received surgical trauma;the electrical acupunctures atBaihuiandDazhuiacupoints on Du channel were performed on the day of model establishment in electric acupuncture group,once a day for 28 days;in fake acupuncture group,sticked the acupuncture needles at the skin ofBaihuiandDazhuipoints,then gived the same electrical acupuncture treatment. On 1,7,14 and 28 days after treatment,the rats of each group were respectively sacrificed,and the brains were collected,then the infarct volume and spinal neuron number were calculated by Nissl staining. Results ①Cerebral infarction volume:No infarcts were found in hypertension group and sham operated group. On 1 day and 7 days after MCAO,the infarct volumes were increased gradually in model group,electric acupuncture group and fake acupuncture group〔infarct volumes on 1 day were(12.36±0.11)%, (12.19±0.15)%,(12.24±0.16)%,and on 7 days were(20.01±0.24)%,(19.54±0.61)%and(19.77±0.25)%, respectively〕,and on 14 days and 28 days after MCAO,the infarct volumes were decreased gradually〔infarct volumes on 14 days were(17.18±0.23)%,(16.96±0.11)%,(17.08±0.62)%,and on 28 days were(14.38±0.21)%, (13.95±0.46)% and(14.29±0.37)%,respectively〕,but the differences among three groups had no statistical significance(all P>0.05). ② Cortex area:On 1 day after MCAO,the differences in the cortical neuron numbers among all groups were not statistically significant(all P>0.05). On 7,14,28 days after MCAO,the cortical neuron number of sham operated group was more than that of hypertension group,but the difference had no statistical significance(all P>0.05). Compared with sham operated group,the cortical neuron number in model group began to increase significantly after 7 days;compared with model group,the cortical neuron number in electric acupuncture group was increased obviously(cell/HP,7 days:75.48±2.41 vs. 68.78±1.42,14 days:61.32±2.60 vs. 48.78±1.41,28 days:53.65±1.46 vs. 28.78±1.21,all P<0.05),while the cortex neuron number of fake acupuncture group was markedly reduced(7 days:67.75±1.43,14 days:47.50±1.25,28 days:27.50±1.25), but the differences had no statistical significance(all P>0.05).③Spinal cord area:On 1,7,14 days after MCAO, the differences of the spinal cord neuron numbers among all groups were not statistically significant(all P>0.05). On 28 days,compared with hypertension group,the cord neuron number of sham operated group was increased,but the difference had no statistical significance(P>0.05). Compared with model group,the cord neuron number in electric acupuncture and fake acupuncture groups was inecreased(cell/HP:21.32±1.60,16.17±1.05 vs. 15.02±1.18),the difference being statistical significant in electric acupuncture group(P<0.05)but no statistical significance in fake acupuncture group(P>0.05). Conclusions Generally,the secondary spinal(cervical part)neuron death occurs after cerebral infarction in rats. The therapeutic action of electric acupuncture may reduce the secondary spinal neuron damage at remote site after cerebral infarction,that is possibly the mechanism of electric acupuncture for the protection of brain in hypertensive rats from I/R injury.

Objective To investigate audiological characteristics of patients with intact tympanic membranes and conductive or mixed hearing loss .Methods A retrospective study was carried out among 30 patients (42 ears) with intact tympanic membranes and conductive or mixed hearing loss who underwent exploratory tympanotomy . The preoperative outcomes of pure tone audiometry ,tympanometry ,resonant frequency of middle ear and temporal bone CT scan were analyzed .Results Among 42 ears ,30 ears with otosclerosis and 12 ears with ossicular chain dis-ruption were confirmed in exploratory tympanotomy ,but only 5 ears showed positive findings in CT scan .The mean thresholds of bone conduction ,air conduction and air -bone gap at frequencies of 0 .5 ,1 and 2 kHz were 27 .5 ± 1 .3 dB HL ,67 .0 ± 1 .8 dB HL ,39 .5 ± 1 .1 dB HL ,respectively .An analysis of tympanometric data of all patients re-vealed that 50% of all ears (21/42) were type A tympanograms ,42 .9% (18/42) were type As tympanograms ,and 7 .1% (3/42) were type Ad tympanograms .The mean of the resonant frequency of the middle ear in otosclerositic patients (1 079 .0 ± 67 .4 Hz) was significantly higher than ossicular chain disruption patients (633 .3 ± 43 .6 Hz) . Conclusion Otosclerosis is the most common in the patients with intact tympanic membranes and conductive or mixed hearing loss .The middle ear resonant frequency of otosclerositic patients is significantly higher than that of ossicular chain disruption patients .

Objective To explore the change of glyoxalase I in type 2 diabetic ocular muscles palsy (DOMP) and its associations with advanced oxidation protein products (AOPP) and oxidative stress. Methods 58 DOMP patients, 50 T2DM and 30 normal controls were enrolled in this study. Levels of blood lipids, fasting blood glucose, hemoglobin A1c, insulin, serum glyoxalase I, AOPP, malondialdehyde (MDA), superoxide dismutase (SOD) and total antioxidant capacity (T-AOC) were measured. Homeostasis model assessment was performed to evaluate the status of insulin resistance (IR). Results Levels of high-density lipoprotein cholesterol, SOD and T-AOC were positively correlated with glyoxalase I and inversely associated to AOPP. Levels of triglycerides , low-density lipoprotein cholesterol , fasting blood glucose , hemoglobin A1c , IR and MDA were negatively correlated with glyoxalase I and positively related to AOPP. AOPP had an inverse association with glyoxalase I (r = -0.823, P < 0.001). Multivariate regression analysis showed that serum levels of glyoxalase I (Sβ = 0.554) and AOPP (Sβ= -0.469) were influencing factors of groups. Conclusion Serum glyoxalase I levels were significantly decreased in DOMP and correlated with AOPP and levels of oxidative stress , which suggest that glyoxalase I could play crucial roles on the development of DOMP.

Objective To investigate the characteristics of auditory brainstem response (ABR) in autistic children of 6 years or younger .Methods A total of 98 children with normal hearing and 22 autistic children were en‐rolled in this study .They were divided into 3 groups :1~2 years age group ,2< ~3 years age group ,and 3< ~6 years age group .The thresholds ,wave latencies and interpeak intervals of ABR were statistically analyzed .Results The ABR thresholds in normal hearing children (and autistic children) in 1~2 years age group ,2< ~3 years age group ,and 3< ~6 years age group were 15 .64 ± 5 .02 (17 .50 ± 5 .35) ,14 .73 ± 4 .85 (17 .81 ± 6 .58) ,and 15 .57 ± 5 .62 (17 .25 ± 6 .97) dB nHL ,respectively .There were no significantly statistical difference between the normal hearing children and autistic children (P>0 .05) .In 80 dB nHL click sound stimulus ,there was also no statistical difference in the values of peak latencies or interpeak intervals of ABR between normal hearing children and autistic children (P> 0 .05) ,whereas ,prolongation of peak latencies or interpeak intervals of ABR occurred in 36 .36%(25 .00% ears) autistic children .The ratio of prolongation of peak latencies of I ,III ,V waves and interpeak inter‐vals of I-III ,III-V ,I-V occurred in 4 .54% ,6 .82% ,13 .64% ,18 .18% ,6 .83% and 15 .91% ,respectively . Conclusion The ABR in some autistic children present abnormality and occur in various forms with the prolongation of interpeak intervals of I -III as the most common .

Objective To investigate the effects of advanced oxidation protein products (AOPP) in type 2 diabetic ocular muscles palsy (DOMP). Methods 58 DOMP patients and 50 type 2 diabetes patients were included in the research. Hemoglobin A1c (HbA1c), blood glucose (FPG), fasting insulin (FINS) and triglycerides (TG), total cholesterol (TC), high-density lipoprotein (HDL), low-density lipoprotein (LDL) were measured and recorded. Homeostasis model assessment was performed to evaluate the status of insulin resistance (HOMA-IR), basal insulin secretion (HOMA-β) and the insulin sensitivity index (ISI). Serum AOPP was measured by enzyme linked immunosorbent assay. Unconditional logistic regression model was used to evaluate the influencing factors of DOMP. Results The DOMP group showed higher levels of plasma AOPP, TG, LDL, FPG, FINS, HbA1c and HOMA-IR, but lower levels of HDL, HOMA-β and ISI than those of the T2DM group. Unconditional logistic regression analysis revealed that AOPP was an independent risk factors for DOMP (OR =3.01, P = 0.002). Conclusion AOPP may be involved in the pathogenesis of DOMP. AOPP could be a useful indicator for monitoring the development of DOMP and for evaluating its severity.

Objective: To analyze the strengths, weaknesses, opportunities and threats of the construction on intelligent vaccination clinics in Zhejiang Province, so as to provide countermeasures for promoting the construction of intelligent vaccination clinics in Zhejiang Province. Methods: By reviewing the annual reports of Zhejiang immunization planning, survey data from Zhejiang Centers for Disease Control and Prevention and Immunization Intelligent Service System, data of human resources of immunization planning, vaccine procurement, construction progress of intelligent vaccination clinics and vaccination were collected. The relevant literature was searched to gather information on the construction standards and norms of intelligent vaccination clinics. The analysis of the strengths, weaknesses, opportunities and threats (SWOT) of the construction of intelligent vaccination clinics was conducted, and corresponding countermeasures and suggestions were proposed. Results: The National Immunization Program reported vaccine rate in Zhejiang Province is more than 99%, and standardized vaccination clinics have been popularized throughout the province. The vaccination staff are professional, and a province-wide intelligent immunization service information system has been established, providing the resources and conditions for the construction of intelligent vaccination clinics. However, there are problems such as low data quality and matching efficiency in vaccination, insufficient data interoperability and sharing, unbalanced regional capabilities in intelligent transformation, and uneven distribution of talent and resources. It is crucial to seize the opportunities presented by the development of big data and artificial intelligence, rely on the regional development of the Internet and health industry, seize the opportunity of rapid growth in demand for intelligent vaccination services and high public acceptance, accelerate the construction of intelligent vaccination clinics, and establish intelligent vaccination service standards as soon as possible. Conclusion We should seize the opportunities presented by the digital reform and development, fully utilize the existing vaccination resources and strengths, address the shortcomings, and accelerate the construction of intelligent vaccination clinics in Zhejiang Province.