Objective To evaluate the clinical value and relationship between the serum levels of the Pro-gastrin-releasing peptide 31-98 (ProGRP) , neuron-specific enolase NSE and the different pathohistology types of lung cancer. Methods 353 lung cancer patients were divided into two groups according to pathohistology types: SCLC group( n = 96), NSCLC group( n = 257). 90 lung benign lesion were taken as control group. ProGRP and neu-ron specific enolase in all patients were detected by ELISA. The levels of the ProGRP, NSE and the clinical value were compared among lung cancer,lung benign lesion patients and the different pathohistology types of lung cancer patients . Results The levels of the ProGRP and NSE of SCLC and NSCLC group were higher obviously than that of lung benign lesion( P <0.01 ). In SCLC diagnosis, the sensitivity, specificity, Youden index and Kappa value of the ProGRP and NSE were O. 7708,0. 9444,0. 7153,0.7111 and 0. 7604,0. 8778 ,0. 6382 ,0. 6355 in the monomial de-tection ; Those indexes above were 0.7604,0. 9667,0.7271 and 0. 7221 in combined assay of ProGRP + NSE( on se-quence test) ; and were O. 8229,0.9000,0. 7229 and 0. 7209 in combined assay of ProGRP + NSE( on parallell test). In NSCLC diagnosis,the above indexes were all lower. Conclusions in SCLC diagnosis,the detection of the serum ProGRP is superior to the detection of NSE, the combined assay of ProGRP + NSE (on parallell test) and Pro-GRP + NSE( on sequence test) are superior to the monomial detection of the ProGRP or NSE,and the combined as-say of ProGRP + NSE(on parallell test) is superiro to ProGRP + NSE(on sequence test) ; The diagnosis value of the monomial arid united detection of ProGRP and NSE to NSCLC is not as expected.

We obtained recombinant ferritin from Dermatophagoides f arinae ,and analyzed the characterization of the pro‐tein .A pair of primers was designed according to the known sequence of ferritin gene .The live mites identified and cultured lo‐cally were picked and the total RNA was extracted .The ferritin gene fragment was amplified by RT‐PCR ,and cloned into pET32a vector ,and then transferred into E .coli Top10 .The target gene obtained from the recombinant plasmid by digestion with Bam HⅠand Hind Ⅲ was connected to the prokaryotic expression vector pET‐32a .The expressed recombinant plasmid containing ferritin gene was constructed by cloning target gene into pET‐32a and transferred into E .coli Bl21 (DE3) .The ex‐pressed recombinant protein was analyzed by SDS‐PAGE ,and was purified by immobilized metal ion affinity chromatography (IMAC) .The ferritin expressed by dust mite was analyzed by the method of bioinformatics .The recombinant plasmid pET32a‐ferritin was constructed .SDS‐PAGE showed a correct molecular weight of the recombinant ferritin protein .After purification by affinity chromatography ,the protein showed only one strip on SDS‐PAGE gel .SDS‐PAGE showed a band at 20 kD .Dust mite ferritin contains 8 serine kinase ,7 threonine kinase ,7 tyrosine kinase ,and 0 histidine kinase phosphorylation sites .Hy‐drophilic region is larger than the hydrophobic region and it is an unstable protein .In conclusion ,the ferritin gene has been cloned and expressed .The purified ferritin has high purity . The study provides a basis for further study of composition and physicochemical properties of house dust mite allergen .

Objective To evaluate the effect of sevoflurane on cognitive function of mice with Alzheimer's disease.Methods Twenty male mice carrying mnutations in amyloid precusor protein (APP) and presenilin 1 genes,weighing 30-40 g,aged 7 months,were divided into either sevoflurane group (group Sev) or control group (group C),with 20 mice in each group.Mice inhaled 3％ sevoflurane for 4 h in group Sev,and mice inhaled 30％ oxygen for 4 h in group C.At 1 month after inhaling sevoflurane or oxygen,the mice underwent continuous multiple-trail inhibitory avoidance training.The mice were then sacrificed and hippocampi were isolated for determination of the number of Aβ plaques (by immunohistochemistry) and expression of APP and Tau (S396) phosphorylation (by Western blot).Results Compared with group C,the memory lateucy was significantly shortened,the number of Aβ plaques was increased,the phosphorylation of Tau (S396) was increased,and the expression of APP was up-regulated in group Sev (P＜0.05).Conclusion Sevoflurane can decrease the cognitive function of mice with Alzheimer's disease.

In this study, 17 kinds of Dendrobium species of Fengdous including 39 individuals were collected from 4 provinces. Mitochondrial gene sequences co I, nad 5, nad 1-intron 2 and chloroplast gene sequences rbcL, matK amd psbA-trnH were amplified from these materials, as well as nrDNA ITS. Furthermore, suitable sequences for identification of Dendrobium species of Fengdous were screened by K-2-P and P-distance. The results showed that during the mentioned 7 sequences, nrDNA ITS, nad 1-intron 2 and psbA-trnH which had a high degree of variability could be used to identify Dendrobium species of Fengdous. However, single fragment could not be used to distinguish D. moniliforme and D. huoshanense. Moreover, compared to other combined fragments, new type combined fragments nrDNA ITS+nad 1-intron 2 was more effective in identifying the original plants of Dendrobium species and could be used to identify D. huoshanense and D. moniliforme. Besides, according to the UPGMA tree constructed with nrDNA ITS+nad 1-intron 2, 3 inspected Dendrobium plants were identified as D. huoshanense, D. moniliforme and D. officinale, respectively. This study identified Dendrobium species of Fengdous by combined fragments nrDNA ITS+nad 1-intron 2 for the first time, which provided a more effective basis for identification of Dendrobium species. And this study will be helpful for regulating the market of Fengdous.

Bletilla striata has been used as traditional Chinese medicine for several centuries. In recent years, the quality and quantity of wild B. striata plants have declined sharply due to habitat deterioration and human over-exploitation. Therefore, it is of great urgency to evaluate and protect B. striata wild plant resource. In this study, sequence-related amplified polymorphism (SRAP) markers were applied to assess the level and pattern of genetic diversity in twelve populations of B. striata. The results showed a high level of genetic diversity (PPB = 90.48%, H = 0.349 4, I = 0.509 6) and moderate genetic differentiation among populations (G(st) = 0.260 9). Based on the unweighted pair-group method with arithmetic average (UPGMA), twelve populations gathered in three clusters. The cluster 1 included four populations. There are Nanjing, Zhenjiang, Xuancheng and Hangzhou. The seven populations which come from Hubei Province, Hunan Province, Jiangxi Province and Guizhou Province belonged to the cluster 2. The cluster 3 only contained Wenshan population. Moreover, Mantel test revealed significant positive correlation between genetic distances and geographic distances (r = 0.632 9; P < 0.000 1). According to the results, we proposed a series of conservation consideration for B. striata.

Objective To investigate the prognosis and efficiency of glucocorticoid and immunosuppressor in the treatment of idiopathic membranous nephropathy(IMN)in children. Methods A retrospective analysis of 35 cases of biopsy - proven membranous nephropathy without secondary factors was performed,who were found present with ne-phrotic proteinuria and admitted to hospital from March 2004 to July 2013,to explore the efficiency of treatment with glucocorticoid and immunosuppressor and its prognosis. Results The 35 IMN cases included 18 boys and 17 girls,and the ratio was 1. 1∶ 1. 0. The mean age at onset was(11. 3 ± 0. 5)years with a range of 3. 0 - 17. 1 years. Five cases with gross hematuria,24 cases present with microscopic hematuria,8 cases with hypertension,1 case with chronic renal insufficiency,and 2 cases were complicated with thrombosis. According to membranous nephropathy staging criteria,9 cases(25. 7% )were in stage Ⅰ,16 cases(45. 7% )in stage Ⅱ,10 cases(28. 6% )in stage Ⅲ;about 94. 3%(33 / 35 cases)had mesangial cells and mesangial matrix with mild to moderate hyperplasia. They were all treated with glucocor-ticoid initially and one of them showed sensitive to flucocorticoid but developed flucocorticoid resistance after relapse, while all the others were flucocorticoid - resistant. Cyclophosphamide A(CsA)was introduced to 17 cases and at least lasted for 3 months,in which 13 cases(76. 5% )reached complete remission and 3 cases reached partial remission, while 1 case didn't achieve remission,and the mean time for proteinuria to disappear was(4. 9 ± 3. 7)months;5 cases were treated with Mycophenolate mefetil( MMF),among which 4 cases reached complete remission in 2 months,4 months,5 months,and 9 months separately,while 1 case reached partial remission. Cyclophosphamide(CTX)was intro-duced to 6 cases,in which the mean cumulative dosage was(91. 2 ± 46. 5)mg/ kg,among them 1 case(87 mg/ kg) reached complete remission,1 case(160 mg/ kg)partial remission,but 4 cases didn't achieve remission. One case reached remission after Rituximab(RTX)was introduced. One case got partial remission after Leflunomide(LEF)was introduced,and the complete remission rate was higher in those treated with combined therapy of glucocorticoid and CsA than those treated with glucocorticoid only(76. 5% vs 12. 5% ,P = 0. 004),but the total efficacy showed no difference (94. 2% vs 62. 5% ,P = 0. 081). The complete remission rate(76. 5% vs 38. 5% ,P = 0. 042)and total efficacy (94. 1% vs 61. 5% ,P = 0. 040)were higher in those with combined therapy of steroid and CsA than those treated with steroid and other immunosuppressor. The complete remission rate(76. 5% vs 16. 7% ,P = 0. 018)and total efficacy (94. 1% vs 33. 3% ,P = 0. 008)were also higher than those treated with steroid and CTX,but the complete remission rate(76. 5% vs 80. 0% ,P = 0. 687)and total efficacy(94. 1% vs 100. 0% ,P = 0. 773)showed no difference com-pared with those treated with steroid and MMF. Conclusions IMN shows glucocorticoid resistance mostly,while CsA had definite efficiency and may be better than CTX. And the efficiency of MMF should be noted.

Objective To summarize the clinical characteristics,diagnosis and management methods and prognosis of hepatorrhagia and splenorrhagia of newborns.Methods A retrospective review of clinical data of neonates with hepatorrhagia and splenorrhagia in perinatal period was performed from June 1,1992 to June 1,2009 in Nanjing Children's Hospital.Results There were twenty-three neonates suffered from hepatorrhagia and splenorrhagia in the perinatal period.There were 12 macrosomias and 6 preterm newborns.Abnormal birth history (65.2%,15/23),including caesarean section,fetal distress,application of vacuum extractor,prolonged labour and precipitate labor,were most commom reasons of hepatorrhagia and splenorrhagia,and birth injuries [47.8% (11/23)]was subsequent.In all cases,14 cases were hepatorrhagia,six were splenorrhagia,three were hepatorrhagia and splenorrhagia simultaneously.Primary early symtoms included low response,sleepiness (91.3%,21/23);jaundice 78.3% (18/23);pallor and anemia 73.9% (17/23);abdominal distension (65.2%,15/23) and so on.Ultrasonography and computed tomography may make a definite diagnosis.Fifteen newborns underwent non-operative treatment and 8 received hemostatic laparotomy.The general mortality was 13.0%(3/23).Conclusions Hepatorrhagia and splenorrhagia of neonates in perinatal period is associated with macrosomias,abnormal birth history and preterm birth,and birth injuries were the major etiological factors.Clinical presentations are nonspecific which maybe asociated with the degree of blood loss.Abdominal ultrasonography is an optimal diagnostic method.Nonoperative management may be successful in hemodynamically stable patients,while immediate intervention,such as laparotomy,is required to control persist bleeding.

Objective:In the operation of esophageal cancer, nano carbon suspension was used for lymph tracing, and the effect of lymph tracing in different regions and its influence on prognosis were analyzed.Methods:From July 2013 to August 2014, 100 patients under esophageal cancer surgery in our department were analyzed. Randomly divided into experimental group and control group, 50 cases each. During the operation, nano carbon suspension was used to trace the lymph nodes in the experimental group, while conventional operation was only used in the control group. The general baseline data, the number of lymph nodes and positive lymph nodes in each region in the two groups were compared, the lymph node metastasis and survival were followed up, and the prognosis of the two groups were compared.Results:There was no significant difference in baseline data between the two groups( P>0.05). The average number of lymph nodes in the experimental group was 18.6 per case, and that in the control group was 15.1 per case. There was significant difference between the two groups( P=0.003). There was no significant difference in the number and degree of metastasis between the two groups( P=0.233, P=0.463). The effect of nano carbon suspension on tracing lymph nodes around the lower mediastinal esophagus was better. In the experimental group, 4.38 lymph nodes were removed in this area on average, 0.34 of which were metastatic lymph nodes. In the control group, 2.52 lymph nodes were removed in each case on average, 0.10 of which were metastatic lymph nodes. The comparison between the two groups was significantly higher than that of the control group( P=0.001, P=0.011 respectively). In the upper mediastinum, abdominal cavity and other mediastinal lymph nodes, there was no significant difference in the number of lymph nodes, the number of lymph nodes and the degree of metastasis between the two groups( P>0.05). The time of lymph node metastasis in the experimental group was significantly later than that in the control group( P=0.048, log rank test). There was no significant difference in survival time between the two groups( P=0.692, log rank test) . Conclusion:The results show that the nano carbon suspension has a good effect on the lymph node tracing during the operation of esophageal cancer, and the effect on the lymph node tracing around the lower mediastinal esophagus is better.In the area close to the lung drainage area, the tracer effect is poor, and it has certain effect in reducing or delaying lymph node metastasis.

Objective To investigate the effect of Anlotinib on proliferation and apoptosis in non-small cell lung cancer(NSCLC)cells and its molecular mechanism.Methods Non-small cell lung cancer cell lines A549 and H1299 were incubated with Anlotinib,miR-16-5p agonist and/or PD-1 overexpression vector respectively.CCK-8 assay and EDU assay were applied to detect the proliferation.Flow cytometry was performed to detect the cell apop-tosis.The relative expression of miR-16-5 p in A549 and H1299 was detected by real-time quantitative polymerase chain reaction(RT-qPCR).The relative protein expression of PD-1 in A549 and H1299 was detected by Western blot assay.The interaction between miR-16-5p and PD-1 was determined by dual luciferase reporter assay.Finally,A549 cell xenograft model was established to assess the effect of Anlotinib on tumor growth in vivo.Results Anlotinib significantly increased miR-16-5p expression and decreased PD-1 expression in A549 cells and H1299 cells,inhibited cell proliferation and promoted apoptosis in a dose-dependent manner(P＜0.05).The highly-expressed miR-16-5p inhibited proliferation and promoted cell apoptosis(P＜0.05).Also,miR-16-5p targeted at PD-1 and negatively regulated PD-1 expression.Knockdown of PD-1 inhibited proliferation and pro-moted cell apoptosis(P＜0.05).PD-1 over-expression reversed the Anlotinib-mediated pro-proliferation and anti-apoptosis of miR-16-5p in A549 cells and H1299 cells(P＜0.05).Anlotinib significantly reduced tumor growth in vivo(P＜0.05).Conclusions Anlotinib may inhibit cell proliferation,anti-apoptosis,and reduce tumor growth for NSCLC,which is involved in miR-16-5p/PD-1 axis.

Objective To study the curative efficacy and safety of single-unit umbilical cord blood transplantation (sUCBT) for malignant hematologic diseases,which is provided by China's public cord blood bank.Methods We retrospectively analyzed 409 cases of malignant hematologic diseases who accepted myeloablative single-unit unrelated donor UCBT without ATG at our center between May 2008 and December 2016.A comparative analysis was made on the total nuclear cells (TNC) of the umbilical cord blood before freezing and after thawing,the cells of CD34+,the recovery rate of cells and the clinical effect of UCBT.Result 409 units of umbilical cord blood used in UCBT respectively came from eight China's public cord blood banks.The average TNC of 409 units of umbilical cord blood before freezing and after the tubular recovery were respectively 18.5 × 108 and 16.34 × 108 (p =0.000).The average recovery rate of the tubular recovery was 88.5％,and there was significant difference among cord blood banks (P =0.000).The average TNC of umbilical cord blood before freezing and transfusion were respectively 18.5 × 108 and 15.86 × 108 (p =0.000).The average recovery rate of umbilical cord blood transfusion was 85.9％,with the difference being significant among cord blood banks (P =0.000).The average number of CD34+ cells before freezing and after the tubular recovery was 11.18 × 106and 8.68 × 106 (p =0.000).The average recovery rate of CD34+ cells after the tubular recovery was 80.75 ％,with the difference being significant among the cord blood banks (P =0.000).At 42nd day after UCBT,the cumulative incidence of neutrophil engraftment was 95.4％,and the median time of the engraftment was 17 days (11-38 days).The cumulative incidence of platelet engraftment at 120th day was 84.6％,and the median time of the engraftment was 36 days (14-93 days).The cumulative incidence of erythrocyte engraftment at 60th day was 92％,and the median time of engraftment was 22 days (9d-60 days).After the umbilical cord blood provided by each bank was used in UCBT,it got the difference in cumulative incidence of engraftment.The P values for cumulative incidence of neutrophil,platelet and erythrocyte engraftment were respectively 0.004,0.01 and 0.000 2,with the differences being statistically significant.At 100th day after UCBT,the cumulative incidence of Ⅱ-Ⅳ and Ⅲ-Ⅳ degrees of acute graft-versus-host disease (aGVHD) was respectively 28.63％ and 15.7％.After umbilical cord blood provided by each bank was used in UCBT,it got the difference in cumulative incidence of aGVHD.There was no significant difference between Ⅱ-Ⅳ and Ⅲ-Ⅳ degrees (P =0.809 and 0.68 respectively).At 3rd year after UCBT,the cumulative incidence of relapse was 15.89％.After umbilical cord blood provided by each bank was used in UCBT,there was no significant difference in the cumulative incidence of relapse (P =0.898).At 3rd year after UCBT,the overall survival (OS) rate and disease free survival (DFS) rate were respectively 66.7％ and 59％.After umbilical cord blood provided by each bank was used in UCBT,it got the difference in OS and DFS.There was no significant difference in OS and DFS (P =0.566 and 0.703 respectively).At 3rd year after sUCBT,the rate of graft-versus-host diseases/relapse-free survival (GRFS) was 54.3％.After umbilical cord blood provided by each bank was used in UCBT,there was no significant difference in the rate of GRFS (P =0.449).Conclusion The umbilical cord blood provided by China's public cord blood bank was used in UCBT.It has a high safety and good efficacy in treating malignant hematologic diseases.But it needs to set up the standardized and normalized quality-control system of umbilical cord blood for China's public cord blood bank.