Complete plastid genomes of Bupleurum chinense DC. and B. boissieuanum H. Wolff, with comparative and phylogenetic analyses of medicinal Bupleurum species / 药学学报

italic>Bupleurum L. (Apiaceae) is an economically important genus, in which many species are of medicinal value. In this study, the complete plastid genomes (plastomes) of B. chinense DC. and B. boissieuanum H. Wolff were sequenced and their characteristics were investigated. Comparative and phylogenetic analyses were conducted with other published Bupleurum plastomes. The complete plastomes of B. chinense and B. boissieuanum were 155 458 and 155 800 bp in length, and both exhibited the typical quadripartite circular structure consisting of a large single copy region (LSC, 85 343 and 85 804 bp), a small single copy region (SSC, 17 495 and 17 410 bp), and a pair of inverted repeat regions (IRa/b, 26 310 and 26 293 bp), respectively. A total of 129 genes, including 84 protein-coding genes, 37 transfer RNA (tRNA) genes, and eight ribosomal RNA (rRNA) genes were identified from each of the two plastomes. Repeat sequences detected were similar in types and distribution patterns, but the numbers were slightly different. Comparative analyses revealed that the Bupleurum plastomes were highly conserved in length, structure, the guanine and cytosine (GC) content, and gene content and order, both intraspecifically and interspecifically, and no obvious expansion or contraction of the inverted repeat regions occurred. Sequence variation was lower within the same species than among different species, noncoding sequences (including intergenic regions and introns) showed a higher divergence than the protein-coding sequences, and sequences in the LSC and SSC regions were more divergent than those in the IR regions. In addition, 11 sequences with higher nucleotide diversity among species were detected in the LSC and SSC regions. All studied Bupleurum species were inferred forming a monophyletic group with a 100% bootstrap value. Bupleurum chinense and B. boissieuanum were phylogenetically closest to B. commelynoideum and B. falcatum, separately, with all three B. chinense accessions clustered into a distinct clade. These results provide genetic information for further species identification, phylogenetic resolution, and will assist in exploration and utilization of medicinal Bupleurum species.

Effect of Arisaema Rhizoma and Pinelliae Pedatisectae Rhizoma on Water Metabolism and Hemorheology in Rats / 中国实验方剂学杂志

Objective:To discuss the effect of Arisaema Rhizoma and Pinelliae Pedatisectae Rhizoma on water metabolism and hemorheolog in rats. Method:A total of 50 healthy SD rats were randomly divided into 5 groups, namely normal group, high-dose Arisaema Rhizoma group (1.2 g·kg·d-1), low-dose Arisaema Rhizoma group (1.2 g·kg·d-1), high-dose Pinelliae Pedatisectae Rhizoma group (1.2 g·kg·d-1) and low dose Pinelliae Pedatisectae Rhizoma group (0.3 g·kg·d-1), with 10 rats in each group. The effects of Arisaema Rhizoma and Pinelliae Pedatisectae Rhizoma on water metabolism and hemorheology were compared, with the micturition index, fecal volume, fecal water content, pathological morphology of submandibular gland, aquaporin (AQP2) content in kidney, adenosine cyclic 3′,5′-(hydrogen phosphate)/current good manufacture practices (cAMP/cGMP) and Na+-K+-ATP enzyme content in serum as the evaluation indexes of water metabolism, and the whole blood viscosity (WBV) and erythrocyte aggregation index (RE) as the evaluation indexes of hemorheolog. Result:Compared with normal group, the fecal volume, fecal water content and AQP2 expression decreased significantly， acinus of submandibular gland were atrophied， micturition index, cAMP/cGMP and Na+-K+-ATP enzyme content in serum were increased significantly， WBV and RE were increased significantly in high-dose Arisaema Rhizoma and Pinelliae Pedatisectae Rhizoma groups（P<0.05）, but with no significant difference between them. Conclusion:High-dose Arisaema Rhizoma and Pinelliae Pedatisectae Rhizoma groups had significant effects on water metabolism and hemorheology in rats, suggesting that attention shall be given to the safety of Arisaema Rhizoma and Pinelliae Pedatisectae Rhizoma. And this study provides a theoretical basis for the rational use of the two traditional Chinese medicines in clinic.

Application of ultrasonic visualization technology in the treatment of war trauma / 第二军医大学学报

In modern war, the advancement of military technology and weapons results in the gathering of severe casualties and increases the difficulty in field rescue, which brings great challenges for medical staff to complete the assessment, classification and rescue tasks in a short time. Ultrasonic examination can easily and quickly screen and diagnose the wounded in battlefield environment or field hospital, and improve the efficiency of classification and treatment. Ultrasonic visualization technology can improve the screening and diagnosis efficiency of war trauma patients with chest, abdomen, craniocerebral and skeletal soft tissue injuries. It also provides convenience for emergency treatment of war trauma patients and improves the emergency treatment effects. This paper reviews the application of ultrasonic visualization technology in the evaluation and emergency treatment of war trauma patients.

Effect of Compound Longmaining on TLR4/MyD88/NF-κB p65 Signaling Pathway in Rats with Acute Myocardial Infarction / 中国实验方剂学杂志

Objective: To investigate the protective effect of compound Longmaining isoprenaline hydrochloride-induced myocardial infarction model and its effect on Toll-like receptor 4(TLR4)/myeloid differentiation factor 88(MyD88)/nuclear transcription factor-kappa B (NF-κB) signaling pathway.Method: Forty-eight male SD rats were randomly divided into 6 groups:normal group,model group,compound salvia miltiorrhiza drop pill group (0.072 9 g·kg-1),and low,medium and high-dose compound Longmaining decoction groups (0.36,0.71,1.43 g·kg-1).The acute myocardial infarction model was induced through subcutaneous injection with isoproterenol.The pathological changes of myocardial tissue were examined by hematoxylin-eosin (HE) staining.The levels of interleukin-1β(IL-1β),interleukin-6(IL-6),tumor necrosis factor-α(TNF-α),monocyte chemotaxis protein-1(MCP-1) and nitrogen (NO) in serum were measured by enzyme linked immunosorbent assay (ELISA).The expression levels of inhibitors of NF-κB kinase subunit-β(IKKβ),NF-κB inhibitor α(IκBα),TLR4,MyD88 and NF-κB p65 were measured by immunohistochemical staining and Western blot.Result: Compared with normal group,the myocardial injury in model group was obvious.The levels of IL-1β,IL-6,TNF-α,MCP-1 and NO in serum increased significantly (PκBα decreased significantly (Pβ,TLR4,MyD88 and NF-κB p65 increased significantly in myocardial tissue (Pβ,IL-6,TNF-α,MCP-1 and NO levels in the serum (PκBα(Pβ,TLR4,MyD88 and NF-κB p65(PConclusion: Compound Longmaining plays a protective effect on acute myocardial infarction by regulatingthe expressions of TLR4/MyD88/NF-κB p65 signaling pathway and relevant inflammatory factors.

Analysis of influencing factors of lymph node metastasis in patients with central and peripheral types of squamous cell lung cancer / 天津医药

Objective To analyze the factors related to the metastasis of lung squamous cell carcinoma in patients with central and peripheral types of lung squamous carcinoma, and provide evidence-based basis for preoperative preparation, intraoperative lymph node treatment choice and postoperative treatment. Methods Clinical data of 142 patients with lung squamous carcinoma who performed operation in Tianjin Medical University Cancer Institute and Hospital from January 2014 to December 2015 were retrospectively analyzed. There were 83 cases of central lung squamous cell carcinoma and 59 cases of peripheral lung squamous carcinoma in these 142 patients. Single factor and multiple factor Logistic regression analysis were used to analyze the clinical data in patients with different type groups, including gender, age, smoking history and smoking index, preoperative CYFRA21-1 level, preoperative predictive lymph nodes metastasis, tumor diameter, tumor type (keratotic and non-keratotic), pleura invasion, tumor embolism and postoperative lymph node metastasis. Results There were 47 cases with lymph node metastasis in 147 patients, including 31 cases of central type metastasis, and 16 cases of peripheral type metastasis. Single factor analysis showed that gender, smoking history, smoking index, preoperative lymph node metastasis assessment, tumor diameter, and non-keratotic type were related to the metastasis of central type of lung squamous cell carcinoma. Pleural invasion and non-keratotic type were associated with peripheral type of pulmonary squamous cell carcinoma. Multivariate Logistic regression analysis showed that more smoking, large tumor diameter and non-keratotic type were independent risk factors for lymph node metastasis of central type of lung squamous cell carcinoma. The non-keratotic type was an independent risk factor for lymph node metastasis of peripheral pulmonary squamous carcinoma. Conclusion Non-keratotic type of squamous cell carcinoma is related to lymph node metastasis especially N1 lymph node metastasis. It is suggested that such patients should be actively treated and colsely followed up after the operation. Patients with more smoking may be more likely to have N2 lymph node metastasis.

Effect of compound Longmaining decoction with different combinations on intestinal absorption of puerarin / 中国中药杂志

To study the effects of compound Longmaining(FFLMN) with different combinations on the intestinal absorption of puerarin. The rat single pass intestinal perfusion model was adopted, and the concentration of puerarin in intestinal samples was determined by HPLC. The effects of different combination groups on the absorption of puerarin in duodenum, jejunum, ileum and colon were investigated. The combined drugs were GG(Puerariae Lobatae Radix), GG-CSL (Puerariae Lobatae Radix compared with Dioscoreae Nipponicae Rhizoma), GG-CX(Puerariae Lobatae Radix compared with Chuanxiong Rhizoma) and FFLMN (compound Longmaining). We found that the absorption rate constant(Ka) and the apparent coefficient(Papp) of puerarin had no significant difference between GG-CSL and FFLMN groups, but significantly higher in GG and GG-CX groups(P<0.05) in the duodenum and ileum. In jejunum and colon, Ka and Papp of puerarin showed significant differences between GG and other groups(P<0.05). At the same time, FFLMN also had significant differences with GG-CSL and GG-CX groups(P<0.05). The results showed that in the whole intestine of rats, FFLMN could significantly promote the absorption of puerarin. In the duodenum and ileum, Dioscoreae Nipponicae Rhizoma played a significant role in promoting absorption of puerarin. In jejunum and colon, Dioscoreae Nipponicae Rhizoma and Chuanxiong Rhizoma have a synergistic effect in promoting absorption of puerarin.

Effect of puerarin in Longmaining formula on pharmacokinetics-pharmacodynamics correlation in rats with myocardial ischemia / 中国中药杂志

To study the pharmacokinetics of puerarin in compound Longmaining(FFLMN) in normal rats and myocardial ischemia rats, and investigate its correlation with anti-myocardial ischemia effect of FFLMN. Models of myocardial ischemia rats were produced by subcutaneous injection of isoproterenol(ISO), then FFLMN extract solution was administered by gavage. Orbital sinus blood sampling was collected at different time points after gavage. HPLC-UV method was applied to determine the concentration of puerarin in plasma, and compare the difference in pharmacokinetics between normal rats and model rats after application of same dose of FFLMN. Meanwhile, microplate reader was used to determine IL-6 and SOD activities in plasma of different time points, and draw dose-effect curve. The results indicated that the pharmacokinetics of puerarin conformed to the two-compartment model in both normal group and myocardial ischemia model group. In the comparison of main pharmacokinetic parameters between two groups: AUC0-∞=(11.451±3.228) mg•h•L⁻¹,AUC0-t=(14.047±3.765) mg•h•L⁻¹, Cmax=(5.623±1.40) mg•L⁻¹ in normal group; AUC0-∞=(68.849±50.396 9) mg•h•L⁻¹, AUC0-t= (58.312±45.802) mg•h•L⁻¹,Cmax=(18.456±7.517) mg•L⁻¹ in treatment group. The SOD level was significantly increased and IL-6 concentration was significantly decreased in plasma, indicating that as compared with the normal group, puerarin in FFLMN had a higher plasma concentration, slower elimination rate and higher bioavailability. Therefore, puerarin concentration in plasma has correlation with the anti-myocardial ischemia effect of FFLMN, which could increase SOD level and inhibit the release of IL-6.

Evaluation of the wound-healing activity and anti-inflammatory activity of aqueous extracts from Acorus calamus l.

In folklore medicine, Acorus calamus has been used as a wound-healing agent for thousands of years; however, there have been few scientific reports on this activity so far. Now, we explored deeply the wound-healing effect of aqueous extracts from the fresh roots and rhizomes of A. calamus in vivo, as well as anti-inflammatory activity in vitro, so as to provide scientific evidence for the traditional application. The wound-healing effect was determined by the image analysis techniques and the histological analysis in the excisional wounding test, and the anti-inflammatory activity was evaluated by the real-time RT-PCR techniques in the lipopolysaccharide-induced RAW 264.7 cells test. Aqueous extracts, administered topically at the dose range from twice to thrice in a day, could enhance significantly the rate of skin wound-healing. Moreover, the extracts could effectively inhibit the mRNA expressions of inflammatory mediators induced by lipopolysaccharide in RAW 264.7 cells. These results showed significantly the wound-healing activity of aqueous extracts in the animal model of excise wound healing, and anti-inflammatory activity in vitro.

Expressions of CLDN1 and insulin-like growth factor 2 are associated with poor prognosis in stage N2 non-small cell lung cancer / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Patients with single station mediastinal lymph node (N2) non-small cell lung cancer (NSCLC) have a better prognosis than those with multilevel N2. The molecular factors which are involved in disease progression remain largely unknown. The purpose of this study was to investigate gene expression differences between single station and multilevel N2 NSCLC and to identify the crucial molecular factors which are associated with progress and prognosis of stage N2 NSCLC.</p><p><b>METHODS</b>Gene expression analysis was performed using Agilent 4×44K Whole Human Genome Oligo Microarray on 10 freshfrozen lymph node tissue samples from single station N2 and paired multilevel N2 NSCLC patients. Real-time reverse transcription (RT)-PCR was used to validate the differential expression of 14 genes selected by cDNA microarray of which four were confirmed. Immunohistochemical staining for these validated genes was performed on formalin-fixed, paraffinembedded tissue samples from 130 cases of stage N2 NSCLC arranged in a high-density tissue microarray.</p><p><b>RESULTS</b>We identified a 14 gene expression signature by comparative analysis of gene expression. Expression of these genes strongly differed between single station and multilevel N2 NSCLC. Four genes (ADAM28, MUC4, CLDN1, and IGF2) correlated with the results of microarray and real-time RT-PCR analysis for the gene-expression data in samples from 56 NSCLC patients. Immunohistochemical staining for these genes in samples from 130 cases of stage N2 NSCLC demonstrated the expression of IGF2 and CLDN1 was negatively correlated with overall survival of stage N2 NSCLC.</p><p><b>CONCLUSIONS</b>Our results suggest that the expression of CLDN1 and IGF2 indicate a poor prognosis in stage N2 NSCLC. Further, CLDN1 and IGF2 may provide potential targeting opportunities in future therapies.</p>

Prognostic analysis of 450 patients with lung squamous cancer / 中华外科杂志

<p><b>OBJECTIVE</b>To study the treatment strategy and prognosis and its affected factors of lung squamous cancer according retrospective analysis.</p><p><b>METHODS</b>Clinic data of 450 lung squamous cancer inpatient cases who were performed complete resection from January 2004 to January 2007, was retrospectively reviewed. There were 363 male and 87 female patients, aged from 31 to 82 years, with a mean of 60.5 years and a median of 62 years.</p><p><b>RESULTS</b>The overall 5-year survival rate was 52.4%. Cox Regression suggested that preoperative N status (χ(2) = 18.969, P = 0.000), N stage (χ(2) = 44.069, P = 0.000) and TNM stage (χ(2) = 63.025, P = 0.000) are independent factors affecting the prognosis. Adjuvant chemotherapy affects the prognosis of stage II-IIIA lung squamous cancer (5-year survival rate: 48.9% vs. 37.7%, χ(2) = 3.946, P = 0.047). Studying the combined therapy of stage IIIA, the chemoradiotherapy group achieved the best survival (48.8%), then single chemotherapy group (35.9%) and no treatment group (28.5%), and the single radiotherapy group achieved the poorest survival rate (11.1%), and there were statistically significant differences among them (χ(2) = 8.397, P = 0.038).</p><p><b>CONCLUSIONS</b>The 5-year survival rate of lung squamous cancer has significantly increased through promoting the standard of operation, especially increasing the standard of lymph node dissection. Adjuvant chemotherapy is benefit for stage II-IIIA patients and combined chemoradiotherapy is the best choice for stage IIIA patients. If preoperative examination suggests mediastinal lymph node's enlargement and fusion, the operation should not be performed.</p>

Osteopontin knockdown suppresses non-small cell lung cancer cell invasion and metastasis / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Osteopontin (OPN) was identified as one of the leading genes that promote the metastasis of malignant tumor. However, the mechanism by which OPN mediates metastasis in non-small cell lung cancer (NSCLC) remains unknown. The aim of the study is to investigate the biological significance and the related molecular mechanism of OPN expression in lung cancer cell line.</p><p><b>METHODS</b>Lentiviral-mediated RNA interference was applied to inhibit OPN expression in metastatic human NSCLC cell line (A549). The invasion, proliferation, and metastasis were evaluated OPN-silenced in A549 cells in vitro and in vivo. The related mechanism was further investigated.</p><p><b>RESULTS</b>Interestingly, OPN knockdown significantly suppressed the invasiveness of A549 cells, but had only a minor effect on the cellular migration and proliferation. Moreover, we demonstrated that OPN knockdown significantly reduced the levels of matrix metalloproteinase (MMP)-2 and urokinase plasminogen activator (uPA), and led to an obvious inhibition of both in vitro invasion and in vivo lung metastasis of A549 cells (P < 0.001).</p><p><b>CONCLUSIONS</b>Our data demonstrate that OPN contributes to A549 cell metastasis by stimulating cell invasion, independent of cellular migration and proliferation. OPN could be a new treatment target of NSCLC.</p>

Epithelial growth factor receptor mutation status to the effective of survival in non-small cell lung cancer after surgery / 中华外科杂志

<p><b>OBJECTIVES</b>To investigate the relationship between the epithelial growth factor receptor (EGFR) mutation status and clinicopathological factors, and to analyze the mutation on the effect in non-small cell lung cancer (NSCLC) after surgery.</p><p><b>METHODS</b>The NSCLC patients who were resected and detected EGFR gene from March 2009 to March 2011 were retrospectively reviewed. The relationship between EGFR mutation status and clinicopathological factors, tumor markers, prognostic was analyzed.</p><p><b>RESULTS</b>The mutation and the wild group had 169 and 214 patients respectively. EGFR mutation in female, non-smoking, adenocarcinoma and less than 60 years old accounted for 63.91%, 61.54%, 88.76% and 62.13% with statistical significance compared with male (χ(2) = 53.490, P = 0.000), smoking (χ(2) = 48.568, P = 0.000), non-adenocarcinoma (χ(2) = 105.560, P = 0.000) and more than 60 years old (χ(2) = 6.057, P = 0.017). Disease free survival (DFS) of the wild group was better than mutation group (χ(2) = 11.329, P = 0.001). In addition, there were some relations between mutation status and excision repair cross complementing (ERCC1) protein, carcinoembryonic antigen (CEA), squamous cell carcinoma (SCC) and Cyfra21-1. ERCC1(+) (χ(2) = 6.739, P = 0.012), SCC(χ(2) = 16.839, P = 0.000) and Cyfra21-1(χ(2) = 6.638, P = 0.013) more than normal value was common in wild group. Increased CEA was common in mutation group (χ(2) = 5.436, P = 0.023).</p><p><b>CONCLUSIONS</b>EGFR mutation is commonly found in female, non-smoking, adenocarcinoma and less than 60 years old NSCLC patients. The wild group obtains better DFS than mutation group. Tumor markers may predict the mutation status, which need further research.</p>

Treatment and prognostic analysis of 1638 patients with non-small cell lung cancer / 中华外科杂志

<p><b>OBJECTIVE</b>To study the prognosis and prognostic factors of non-small-cell lung carcinoma (NSCLC) according the new TNM stage system.</p><p><b>METHODS</b>Clinic data of 1638 inpatient cases admitted from January 2001 to January 2005 were retrospectively reviewed. There were 1083 male and 555 female patients in the study and the average age was 59.5 years. All the patients received surgical procedures.</p><p><b>RESULTS</b>The overall 1, 3, 5-year survival rate was 80.0%, 52.3%, 39.0%. The main prognostic factors were bronchial stump, operation type, T stage, N stage, the number of lymph nodes (LNs) in lymph nodes dissection (1 - 10, 11 - 20, and > 20), overall N stations (< 4 and ≥ 4) and postoperative radiotherapy (all P < 0.05). Cox regression suggested that T stage (P = 0.000), N stage (P = 0.000), operation type (P = 0.001) and LNs (P = 0.013) were independent factors affecting the prognosis.</p><p><b>CONCLUSIONS</b>The overall survival rate of NSCLC is poor. T stage, N stage, operation type and LNs are independent factors affecting the prognosis.</p>

Up-regulation of the expression of S100A8 and S100A9 in lung adenocarcinoma and its correlation with inflammation and other clinical features / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>S100A8 and S100A9 are two members of the S100 protein family characterized by the presence of two Ca2+-binding sites of the EF-hand type. Previous studies suggested that the whole S100 family displays significant functions in tumor growth, progression and invasion. This study aimed to determine the expression of the two indices of the family, S100A8 and S100A9, in lung cancer tissues and normal lung tissues and its correlation with clinical features.</p><p><b>METHODS</b>A total of 60 cases with a variety of clinical data that were diagnosed with different histological subtypes of lung cancer were investigated. Semi-quantitative reverse transcriptase-PCR (Sq-Rt-PCR) and immunohistochemical staining of cancer, adjacent and peripheral lung tissues were executed to distinguish the expression patterns of S100A8 and S100A9 and to further clarify their correlation with clinical features.</p><p><b>RESULTS</b>Immunohistochemical staining of both proteins showed a significant up-regulation in lung cancer tissue (S100A8, S100A9, P<0.0001), and PCR revealed that the levels of S100A8 and S100A9 expression were significantly higher in lung cancer tissues (S100A8 P=0.002/0.004; S100A9 P=0.022/0.026). The higher expression was found to be correlated with the clinical characteristics of adenocarcinoma, inflammation and stage IV lesion.</p><p><b>CONCLUSIONS</b>S100A8, S100A9 up-regulation was found in the lung adenocarcinoma and end stage lung cancer tissue, the correlation of which with their higher expression in inflammatory lung tissues may indicate the collaborative effect of inflammation on the progression of cancer.</p>

A clinical and prognostic retrospective analysis of IIIA-N2 non-small cell lung cancer / 中华外科杂志

<p><b>OBJECTIVES</b>To analyze the clinical conditions of postoperative patients with IIIA-N2 non-small cell lung cancer (NSCLC) and the prognostic factors related with survival of NSCLC, and to investigate the influence of operation and therapy on prognosis.</p><p><b>METHODS</b>Clinical data of 657 inpatient cases with IIIA-N2 NSCLC admitted from January 2000 to December 2005 was retrospectively reviewed. The Kaplan-Meier method was used for survival analysis. The Log-rank law was applied to analyze the relationship between the variables and the prognosis in monovariate analysis, while Cox proportional hazard regression model was used to make multivariate analysis.</p><p><b>RESULTS</b>The 1-, 3-and 5-year accumulative survival rates of the operative patience were 64.4%, 26.0% and 17.9%, respectively. The median survival time was 18 months. In monovariate analysis, the main unfavorable factors that affect life span involve were the diameter of tumor, T stage, skip metastasis of N2 lymph node, the number of metastatic lymph nodes, the metastasis of subcarinal lymph nodes, adjuvant chemotherapy, the cycle of adjuvant chemotherapy, postoperative radiotherapy, and the modality of therapy (the effect of naive surgery was disappointed, while the prognosis of the patients with adjuvant chemoradiotherapy was better than those with chemotherapy alone). A multivariate analysis using Cox regression identified 5 factors of prognosis: the diameter of tumor (P = 0.001), the metastasis of subcarinal lymph nodes (P = 0.019), the number of metastatic lymph nodes (P = 0.006), the cycle of adjuvant chemotherapy (P = 0.007), postoperative radiotherapy (P = 0.055), and adjuvant chemoradiotherapy (P = 0.026).</p><p><b>CONCLUSIONS</b>The 5-year survival rate of the patients with IIIA-N2 Non-small cell lung cancer is poor. Tumor size, the metastasis of subcarinal lymph nodes, the number of metastatic LNs, the cycle of adjuvant chemotherapy, and postoperative radiotherapy have an effect on the prognosis. The prognosis of postoperative patients with single-level N2 and multi-level N2 disease is similar, and the key point of survival is the number of nodes involved. The therapeutic effect of patience given adjuvant chemoradiotherapy is superior to those treated with adjuvant chemotherapy.</p>

Clinical and prognostic analysis of skip N2 metastases in stage IIIA-N2 non-small cell lung cancer / 中华外科杂志

<p><b>OBJECTIVE</b>To investigate the clinicopathologic factors and the distribution pattern of N2 lymph nodes, to analyze the relationship between the survival rate and skip metastasis of non-small cell lung cancer (NSCLC) patients.</p><p><b>METHODS</b>The clinical data of 478 patients with a pN2 stage who underwent resection for non-small cell lung cancer from January 2000 to December 2004 was retrospectively analyzed. Skip group and non-skip group were defined. Characteristics of tumors, ganglionar involvement and survival were analyzed in both groups.</p><p><b>RESULTS</b>The incidence rate of skip N2 metastasis in stage IIIA-N2 NSCLC patients was 40.6%, which was correlated to sex, smoking and the type of histology (P < 0.05). Squamous carcinoma was the main type of skip group (chi² = 7.602, P = 0.022). The frequency of skip metastasis was higher in patients with a primary tumor in the upper lobe (57%) compared to the lower lobe (43%) (chi² = 5.097, P = 0.024). Superior nodes were more frequently involved by skip group (chi² = 7.046, P = 0.030). Moreover, the relationship between the primary tumor location and N2 positive lymph nodes were described as follows: right upper lobe cancer displayed skip-N2 nodal metastasis mostly in the 2nd, 3rd and 4th station, right middle and lower lobe mostly in the 7th station, left upper lobe mostly in the 5th and 6th station (71.7%), and left lower lobe mostly in the 7th and 9th station. The 5-year survival rate of pN2 patients with skip metastasis was 22.1% compared to 13.6% in patients with involvement of N1 and N2 nodes (P = 0.001). Survival analysis showed that skip N2 metastasis was an independent risk factor of stage IIIA NSCLC.</p><p><b>CONCLUSIONS</b>The frequency of skip metastasis was higher in patients with a primary tumor in the upper lobe and in the superior nodes. Skip metastasis is an independent prognostic factor of survival. The presence of skip metastasis seems to be a unique subgroup of pN2 disease in NSCLC.</p>

Study on the effect of bone-cement interface with bone cement oscillator / 中华外科杂志

<p><b>OBJECTIVE</b>To evaluate the effect on increasing bone cement-bone interface micro-gomphosis intensity with bone cement oscillator.</p><p><b>METHODS</b>One hundred femoral bones of adult pig were randomly divided into 6 groups: oscillating group (A1) and control group (A2) of anti-tensile force, oscillating group (B1) and control group (B2) of anti-pressure (n = 20 in each group), oscillating group (C1) and control group (C2) of imaging (n = 10 in each group). Mechanics and CT test was performed, micro-gomphosis intensity of bone cement-bone interface between oscillating group and control group was compared.</p><p><b>RESULTS</b>Mechanics and CT test showed bone cement-bone interface micro-gomphosis intensity in oscillating group was significantly stronger than control group (P < 0.01).</p><p><b>CONCLUSION</b>Bone cement oscillator can significantly increase micro-gomphosis intensity of bone-cement interface, and reduce long-term aseptic loosening of artificial prostheses.</p>

Clinical significance of CD4+ CD25+ regulatory T-cells detection in tumor-draining lymph nodes of nonsmall cell lung cancer patients / 中华肿瘤杂志

<p><b>OBJECTIVE</b>To evaluate the distribution of CD4+ CD25+ regulatory T-cells (T-regs) in tumor-draining lymph nodes (TDLN) in patients with non-small cell lung caner (NSCLC), and to investigate the effect of CD4+ CD25+ T regulatory cells on the immune status of TDLN and the progression of NSCLC.</p><p><b>METHODS</b>Regional tumor-draining lymph nodes of 53 NSCLC patients were resected during the operation. The percentage of CD4+ CD25+ T-regs as a subset of CD4+ T cells and CD8+ T cells were detected by immunofluorescence and regular immunohistochemistry, respectively. The level of cytokines TGF-beta1 and IL-10 was detected by real time quantitative RT-PCR.</p><p><b>RESULTS</b>CD4+ CD25+ T-regs in tumor-infiltrating lymph nodes from the patients with NSCLC accounted for 28.80% +/- 8.06% of total CD4+ T cells, and were significantly increased comparing with that (15.48% +/- 4.66%) in the tumor-free lymph nodes (P < 0.01). The percentage of CD4+ CD25+ T-regs in TDLN of NSCLC patients was negatively correlated with the amount of CD8+ T cells within the lymph nodes (r = -0. 756, P < 0.001), but positively correlated with the level of TGF-beta1 (r = 0.645, P < 0.001) and IL-10 (r = 0.769, P < 0.001). It also increased as NSCLC getting progressed, which was 30.42% +/- 7.47% in stage III versus 16.22% +/- 4.88% in stage I and III; 32.58% +/- 7.52% in N2 versus 22.76% +/- 4.67% in N1, with a significant difference between the two groups, respectively (P < 0.01).</p><p><b>CONCLUSION</b>The population of CD4+ CD25+ T regulatory cells in tumor-draining lymph nodes in patients with non-small cell lung caner is positively correlated with the progression and infiltration of lung cancer, which might provide new immunologic method to evaluate the progression and prognosis of non-small cell lung caner. The outcomes of biotherapy for NSCLC may be improved in the future through regulating the CD4+ CD25+ T regulatory cells.</p>

Generation of T cell-mediated antitumor response in vitro by autologous dendritic cells pulsed with tumor lysates in patients with non-small cell lung cancer / 中华肿瘤杂志

<p><b>OBJECTIVE</b>To investigate whether dendritic cells pulsed with whole tumor lysates (WTL) could in vitro elicit antitumor T cell responses in patients with non-small-cell lung cancer (NSCLC).</p><p><b>METHODS</b>Monocyte-derived immature DCs (imDCs) generated in the presence of human recombinant granulocyte-macrophage colony stimulating factor and interleukin-4 from peripheral blood mononuclear cell of NSCLC patients, and then were induced to mature by pulsing autologous WTL (DCs/WTL) or by the addition of TNF-alpha(TNF/DCs). FACS and MLR assay were used to monitor their phenotypic changes and capacity to stimulate allogeneic and autologous T cell proliferation. DCs/WTL activated with TNF-alpha (* DCs/WTL) were cocultured in vitro with autologous T cells for eliciting antitumor CTLs. T cell mediated antitumor responses were measured by IFN-gamma enzyme-linked immunospot (ELISPOT) assay for WTL-specific IFN-gamma releasing T cells and by lactate dehydrogenase release (LDH) assay for lysis of autologous tumor cells, respectively.</p><p><b>RESULTS</b>When monocytes-derived imDCs from the patients with NSCLC (n = 10) were pulsed with autologous WTL for a day at 30 microg total protein of WTL per 10(6) DCs/ml, this led to up-regulation of CD1a, CD83 and CD86 as well as HLA-DR, and also led to marked stimulation of allogeneic T cell proliferating activity, which was comparable to that of TNF/DCs. However, their capacity of stimulating autologous T cell proliferation in vitro was significantly more potent than those of TNF/DCs (P < 0.05). The numbers of WTL-specific IFN-gamma releasing T cells in 1/3 cultures after one week exposure to * DCs/WTL was increased significantly compared with those pulsing with TNF/DCs plus IL-2 or IL-2 alone (P = 0.05). T cells derived by priming of non-adherent PBMCs with * DCs/WTL after 14 days in vitro stimulation were significantly more responsive to autologous tumor cells compared with LAK (n = 3, P < 0.05), but its cytotoxicity against K562 cells was also comparable to LAK cells.</p><p><b>CONCLUSION</b>Monocyte-derived DCs from NSCLC patients could serve as functional APC. The * DCs/WTL may effectively elicit T cell-mediated antitumor response in vitro and enhance NK killing activity.</p>

Experimental study of the specific immunoresponse induced by p53 modified dendritic cells / 中华血液学杂志

<p><b>OBJECTIVE</b>To explore the specific cytotoxic T lymphocyte (CTL) induced by dendritic cells (DC), which were transfected by the plasmid pC53-SN3 encoding p53 gene.</p><p><b>METHODS</b>DC derived from HLA-A2(+) mononuclear cells of the 24-lung cancer patients was transfected with the plasmid pC53-SN3 by lipofectamine and then co-cultured with auto-unpurified T cells to induce potent CTL (T-pC53-SN3). The cytolysis of specific CTL against Calu-6, a HLA-A2(+) human lung cancer cell line, was measured by using lactate dehydrogenase (LDH) releasing assay.</p><p><b>RESULTS</b>The expression of CD(1a) and CD(83), the correlative markers of DC, increased apparently after transfected with plasmid pC53-SN3, the expression rate was (5.45 +/- 0.89)% and (3.26 +/- 0.47)% versus (52.15 +/- 11.56)% and (25.78 +/- 12.35)%. CD(14) decreased apparently, but other DC correlative markers of CD(1a), CD(40), CD(86), and HLA-DR remained almost the same as that before transfection. Compared with T-IL-2, the CTL derived from PBMNC stimulated by IL-2 (100 U/ml), the cytolytic activity of T-pC53-SN3 against Calu-6 cell line showed a significant increase, but cytolytic activity was 56.79 +/- 15.67 and 39.33 +/- 9.88, respectively, when effect cells: target cells was 10:1. The expression of the CD(8), CD(69), and CD(45)RO/CD(8) of T-pC53-SN3 cells increased significantly, but that of CD(3), CD(4), CD(86), ect, was not significantly different from those of T-pCMV-neo.</p><p><b>CONCLUSIONS</b>It showed that DC transfected by p53 gene could induce potent HLA-A(2) restrictive CTL to kill tumor cell efficiently.</p>