Objective To establish a novel method for detecting circulating tumor cells (CTCs) in phripheral blood of lung cancer patients with high sensitivity and specificity.Methods Experimental study.42 cases of initial treatment patient who underwent resection and diagnosed to be non-small cell lung cancer by biopsy were studied,including 7 patients at stage Ⅰ,9 patients at stage Ⅱ,16 patients at stage Ⅲ and 10 patients at stage Ⅳ.As a control group,20 cases of healthy volunteers were selected.A series of experiments was conducted to determine the efficiency of tumor cells isolation,in which varied concentration (50,100,200,500,1000 cells) of A549 cells spiked into 2 ml peripheral blood drawn from healthy donors.The blood was removed of unwanted erythrocytes by lysis buffer,and made the rest of nucleated cells incubate with anti-EpCAM magnetic beads,then separated and enriched by a specific detector.All epithelia cells were retained on a slide because of a magnetic force and identified by H&E staining protocol.On the basis of cell recovery rate we calculated the sensitivity of tumor cells isolation.20 blood samples taken from healthy individuals were also detected to validate the specificity of this method.Samples of 42 patients with lung cancer were assayed for CTCs detection by above method.The correction of CTCs quantity with the patients' clinical features,for example,ages,gender,clinical stage,tumor size was analyzed in lung cancer patients by chi-square statistics.The correction of recovery cells with the spiked cells were assayed by linear correlation.Results The recovery rate was ranging from 68％ to 82％ by spiking varying numbers of A549 lung cancer cells into 2ml blood samples of healthy volunteers.Regression analysis of number of recovered vs.spiked A549 cells yielded a regression equation of Y =0.6419X + 8.8875.The number of CTCs detected has signification correlate with the cells spiked (R2 =0.9916,P ＜ 0.05),Eighteen of the 42 patients (43％) were found have CTCs in peripheral blood.The detection rate of lung cancer cells was 0 at stage Ⅰ,the detection rate of lung cancer cells was 11.1％ at stage Ⅱ,the detection rate of lung cancer cells was 62.5％ at stage Ⅲ and the detection rate of lung cancer cells was 70％ at stage Ⅳ.The positive rate of CTCs has no signification correlate with ages and gender of patients and tumor size (P ＞ 0.05),has signification with the clinical stage (P ＜ 0.05).None of the peripheral blood samples of the 20 healthy subjects analyzed was found to have CTCs.Conclusions This novel immunomagnetic separation technology is a sensitive and specific method,which provides a new tool allowing for feasible and specific detection of CTCs in lung cancer patients.The level of CTCs increases with the clinical stage and tumor size increased,which has important value to discover the early stage micrometastasis and redefine the clinical stage.But further multicenter and large sample clinical research are needed to confirm its clinical value.

Objective　To detect the location of pathogens in myocardium using in situ RT-PCR technique in order to study the pathogenetic course of the myocardial lesion induced by CoxB3m virus infection in mice. Methods　(1) Thirty and fifty Balb/c mice were used respectively to establish the acute and chronic CoxB3m infected models, with another 25 healthy mice as the controls; (2) KS400 image analysis system (Germany) was used to measure the cardiac chamber area and the left ventricular wall thickness of the chronic infected mice and the controls; (3) CoxB3m virus in myocardial tissue was detected using in situ RT-PCR by direct incorporated technique which employed nucleotide labeling by anti-digoxin antibody and bonded with alkaline phosphatase (anti-dig-AKP method). Results　Picture analysis indicated that the left ventricular chamber area was enlarged and the left ventricular wall was thinner in the chronic repeated virus infected models than those of the controls. With in situ RT-PCR, positive signals for Coxsackie virus B3m RNA were detected not only in the myocardium of the acute Balb/c mice models but also in the myocardium of the chronic mice models. Conclusion　Coxsackie virus B3m is able to induce pathologic lesions by exhibiting positive CVB-RNA signals in both acute and chronic models in mice. In the chronic experimental models, the cardiac chamber is enlarged while the ventricular wall is thinned which demonstrates the association with persistent infection of Coxsackie virus B3m virus.

Objective To explore the influences of the individual cervical hyperextension position on the hemodynamics of vertebral artery in patients with thyroidectomy.Methods According to the random number table,80 participants were divided into the intervention group (n =40)and the control group (n =40).The intervention group used individual cervical hyperextension position while the control group used conventional cervical hyperextension position.The hemodynamics changes of the bilateral vertebral artery for C4 ~5 and the incidence rates for the posture symptom of thyroid surgery were compared between the two groups.Results Both the blood flow rates and volumes of the bilateral vertebral artery were lower in the control group compared with those in the intervention group (P <0.05).The incidence rates for the posture symptom of thyroid surgery were significantly reduced in the intervention group (P <0.01 ).Conclusions The individual cervical hyperextension position can effectively improve the hemodynamics of vertebral artery and reduce the incidence for the posture symptom of thyroid surgery.

Objective:To compare the dosimetric characteristics of helical tomotherapy (HT) and volumetric modulated arc therapy (VMAT) after left breast conserving surgery.Methods:Twenty-four patients with left breast cancer after breast-conserving surgery who were admitted to the Department of Radiation Oncology of Tumor Hospital of Yunnan Province from May 2016 to May 2019 were selected. The HT plan and the VMAT plan were designed for the same patient. The target dose and the dose volume parameters of organs at risk were compared and analyzed in the two radiotherapy plans.Results:There were significant differences in the D 2% [(59.68±0.46) Gy vs. (60.06±0.20) Gy, t=-4.229, P<0.001], D 98% [(57.46±0.44) Gy vs. (57.20±0.07) Gy, t=2.912, P<0.001], conformity index (CI) (0.80±0.05 vs. 0.76±0.04, t=4.079, P<0.001) and homogeneity index (HI) (0.04±0.01 vs. 0.05±0.00, t=-5.505, P<0.001) of the planning gross tumor volume (PGTV) between the HT and VMAT plans. However, there was no significant difference in the D 50% [(58.77±0.46) Gy vs. (58.75±0.11) Gy, t=0.179, P=0.859]. There were significant differences in the D 50% [(51.99±0.39) Gy vs. (52.39±0.36) Gy, t=-5.278, P<0.001], D 98% [(49.46±0.29) Gy vs. (48.35±0.46) Gy, t=9.538, P<0.001] and HI (0.19±0.01 vs. 0.21±0.01, t=-7.538, P<0.001) of the planned target volume (PTV) between the two plans. However, there were no significant differences in the D 2% [(59.13±0.64) Gy vs. (59.09±0.46) Gy, t=0.511, P=0.614] and CI (0.83±0.04 vs. 0.82±0.04, t=1.637, P=0.115). In terms of organs at risk, there were significant differences in the V 5 [(57.90±1.42)% vs. (52.40±5.74)%, t=4.812, P<0.001], V 20 [(22.40±2.17)% vs. (18.40±3.16)%, t=5.573, P<0.001] and D mean [(12.71±0.55) Gy vs. (11.46±1.26) Gy, t=4.963, P<0.001] of left lung, D mean of right lung [(3.42±0.27) Gy vs. (2.49±0.24) Gy, t=13.310, P<0.001], D mean of right breast [(4.41±0.50) Gy vs. (3.12±0.65) Gy, t=10.326, P<0.001], V 30 [(0.55±0.37)% vs. (1.24±1.11)%, t=-4.020, P=0.001] and D mean of heart [(4.68±0.62) Gy vs. (3.83±0.88) Gy, t=7.335, P<0.001], D mean of left atrium [(2.53±0.31) Gy vs. (2.16±0.28) Gy, t=5.488, P<0.001], D mean of right atrium [(2.77±0.43) Gy vs. (2.20±0.30) Gy, t=7.103, P<0.001], D mean of right ventricle [(5.10±0.72) Gy vs. (3.72±0.94) Gy, t=9.802, P<0.001] and D 2% of spinal cord [(14.79±2.73) Gy vs. (5.42±2.23) Gy, t=14.788, P<0.001] between HT and VMAT plans. There was no significant difference in the D mean of left ventricle [(5.10±1.19) Gy vs. (4.80±1.54) Gy, t=1.250, P=0.224]. Conclusion:Both the HT plan and the VMAT plan can meet the treatment requirements. The HT plan can provide better target area conformity and dose uniformity. The VMAT plan has more advantages in terms of organs at risk. The HT plan shows an advantage only in exposure to high-dose area.

Objective:To study the distribution and antibiotics resistance of the main pathogens of neonatal purulent meningitis in different regions of China.Methods:A retrospective descriptive clinical epidemiological study was conducted in children with neonatal purulent meningitis which admitted to 18 tertiary hospitals in different regions of China between January 2015 to December 2019. The test results of blood and cerebrospinal fluid, and drug sensitivity test results of the main pathogens were collected. The distributions of pathogenic bacteria in children with neonatal purulent meningitis in preterm and term infants, early and late onset infants, in Zhejiang Province and other regions outside Zhejiang Province, and in Wenzhou region and other regions of Zhejiang Province were analyzed. The chi-square test was used for statistical analysis.Results:A total of 210 neonatal purulent meningitis cases were collected. The common pathogens were Escherichia coli ( E. coli)(41.4%(87/210)) and Streptococcus agalactiae ( S. agalactiae)(27.1%(57/210)). The proportion of Gram-negative bacteria in preterm infants (77.6%(45/58)) with neonatal purulent meningitis was higher than that in term infants (47.4%(72/152)), and the difference was statistically significant ( χ2=15.54, P=0.001). There were no significant differences in the constituent ratios of E. coli (36.5%(31/85) vs 44.8%(56/125)) and S. agalactiae (24.7%(21/85) vs 28.8%(36/125)) between early onset and late onset cases (both P>0.05). The most common pathogen was E. coli in different regions, with 46.7%(64/137) in Zhejiang Province and 31.5%(23/73) in other regions outside Zhejiang Province. In Zhejiang Province, S. agalactiae was detected in 49 out of 137 cases (35.8%), which was significantly higher than other regions outside Zhejiang Province (11.0%(8/73)). The proportions of Klebsiella pneumoniae, and coagulase-negative Staphylococcus in other regions outside Zhejiang Province (17.8%(13/73) and 16.4%(12/73)) were both higher than those in Zhejiang Province (2.9%(4/137) and 5.1%(7/137)). The differences were all statistically significant ( χ2=14.82, 12.26 and 7.43, respectively, all P<0.05). The proportion of Gram-positive bacteria in Wenzhou City (60.8%(31/51)) was higher than that in other regions in Zhejiang Province (38.4%(33/86)), and the difference was statistically significant ( χ2=6.46, P=0.011). E. coli was sensitive to meropenem (0/45), and 74.4%(32/43) of them were resistant to ampicillin. E. coli had different degrees of resistance to other common cephalosporins, among which, cefotaxime had the highest resistance rate of 41.8%(23/55), followed by ceftriaxone (32.4%(23/71)). S. agalactiae was sensitive to penicillin, vancomycin and linezolid. Conclusions:The composition ratios of pathogenic bacteria of neonatal purulent meningitis are different in different regions of China. The most common pathogen is E. coli, which is sensitive to meropenem, while it has different degrees of resistance to other common cephalosporins, especially to cefotaxime.

Objective: To evaluate the safety and effectiveness of repeated transcranial magnetic stimulation (rTMS) for treating non-fluent aphasia after stroke. Methods: Forty-five stroke survivors with non-fluent aphasia were randomly divided into a 0.5 Hz group, a 1 Hz group and a sham group, each of 15. In addition to routine linguistic training, the three groups were given rTMS over the inferior frontal gyrus of the right hemisphere at the corresponding frequency or sham stimulation. Before as well as 5 and 10 days after the treatment, all of the subjects were evaluated using the Chinese version of the Western Aphasia Battery (WAB). The occurrence of adverse events was also observed. Results: Before treatment, no significant differences were observed in the groups′ average aphasia ratio, spontaneous speech, listening comprehension, retelling and naming using the WAB. After 5 and 10 days significant increases were observed in the average WAB scores of all three groups, but the listening comprehension of the 0.5 Hz group was significantly better than that of the sham group 10 days later, as was the spontaneous speech of the 1 Hz group. Conclusion rTMS at either 1 Hz or 0.5 Hz can improve the linguistic functioning of stroke survivors with non-fluent aphasia. Both 0.5 Hz and 1 Hz rTMS are safe, but the latter is more effective.