Sphingosine kinase 1 (SphK1) plays critical roles in cell biological functions. Here we investigated the effects of SphK1 inhibitor SKI II on hepatoma HepG2 cell cycle progression and invasion. Cell survival was determined by SRB assay, cell cycle progression was assayed by flow cytometry, the ability of cell invasion was measured by Matrigel-Transwell assay and protein expression was detected by Western blotting. The results showed that SKI II markedly inhibited HepG2 cell survival in a dose-dependent manner, induced G1 phase arrest in HepG2 cell and inhibited cell invasion. SKI II markedly decreased the expressions of G1-phase-related proteins CDK2, CDK4 and Cdc2 and the levels of cell invasion-associated proteins MMP2 and MMP9. The results showed that SKI II inhibited cell cycle progression and cell invasion, implying SphK1 as a potential target for hepatoma treatment.

Objective: To explore the effects of different concentrations of glucose solution on the survival of Aedes albopictus and Culex pipiens pallens larvae, the attraction to mosquitoes and egg-laying behaviors, so as to provide the reference for developing mosquito control technology based on sugar bait. Methods: White porcelain bowls were filled with 100 mL of 3%, 5%, 8%, 10% and 15% glucose solutions. Ten of fourth instar larvae of Aedes albopictus or Culex pipiens pallens were added to each bowl, and the survival of larvae was recorded after 2, 4, 6, 24, 48 and 72 hours. Egg-laying cups containing 5%, 8% and 15% glucose solution were put in mosquito cages containing fully blooded female mosquitoes of Aedes albopictus and Culex pipiens pallens (50 mosquitoes each), and the total number of eggs laid in 72 hours was observed. The analogous site room was filled with fully blooded and starved female mosquitoes of Aedes albopictus and Culex pipiens pallens (100 mosquitoes each), and simple mosquito control buckets containing 5% and 8% glucose solution and black sticky insect plates. The number of mosquitoes and eggs was observed after 6 days. All the above experiments were repeated 3 times using dechlorinated water as the control. Results: The 72 hour corrected mortality rates of Aedes albopictus and Culex pipiens pallens larvae gradually increased with the increase of glucose concentration. The glucose solution with 5% and higher concentrations was not suitable for mosquito larvae to survive. The attraction of egg-laying behaviors to Aedes albopictus and Culex pipiens pallens gradually decreased with the increase of glucose concentration. The effects were similar between 5% and 8% glucose solution, with the averages of 686.67 and 682.33 eggs for Aedes albopictus, and 3.00 and 2.33 egg rafts for Culex pipiens pallens. In analogous site room, there were 93.33, 105.00 and 130.33 adult mosquitoes captured on average in the control group, 5% and 8% glucose solution groups, respectively, with 8% glucose solution group more attractive to adult mosquitoes than the control group (F=3.283, P=0.030); there were 70.33, 55.33 and 63.00 Aedes albopictus eggs (eggs counts+larvae counts) on average, respectively, with statistically significant differences among the three groups (H=6.761, P=0.034). Conclusion Glucose solution with concentration of 5% or higher can effectively inhibit the survival of Aedes albopictus and Culex pipiens pallens larvae, and attractive to adult mosquitoes and egg-laying behavoirs.

Objective: To analyze the genetic diversity of Aedes albopictus populations in the coastal areas of southern China by using the microsatellite markers to provide a basis for the control of vectors. Methods: Genetic diversity and clustering analysis of Aedes albopictus populations were studied in the 7 microsatellite loci, in Hangzhou, Ningbo and Yiwu of Zhejiang province, Longyan of Fujian province, Guangzhou of Guangdong province, Nanning of Guangxi Zhuang Autonomous Region and Haikou of Hainan province. Results: Numbers of different alleles (5.429-7.571), effective alleles (2.897-3.632), allele richness (5.236-7.170) and expected heterozygosity (0.538- 0.637) were detected from each of the Aedes albopictus population by using 7 microsatellite markers. The inbreeding coefficients appeared as 0.008-0.332, with heterozygote deficiency, in these populations. Fixation index of the whole populations was 0.058, suggesting that the genetic variation among the 7 populations was 5.8%. Data from the Neighbor-Joining clustering analysis showed that populations from Hangzhou and Yiwu belonged to one branch while Longyan and Guangzhou populations constituted another branch. Aedes albopictus populations of Nanning and Haikou showed great genetic variation but formed a single branch. Bayesian analysis on Aedes albopictus populations showed that the possible number of clusters was 3. Conclusions Based on 7 microsatellite loci, relatively high genetic diversity and medium level of genetic differentiation that increasing with the geographical distances, were found in these Aedes albopictus populations, from the coastal areas in southern China.

Objective: To apply DNA barcoding to identifying the rodents in Zhejiang Province. Methods : Rodents were captured from Jiashan,Longyou,Yunhe and Ninghai counties in Zhejiang Province. The DNA was extracted from ears of rodent samples,and was amplified and sequenced with mitochondrial cytochrome C oxidase subunit I（COI）genes. The obtained sequences were compared with the related sequences in GenBank,and neighbour-joining evolutionary tree was constructed. Then the results by DNA barcoding and by morphological identification were compared. Results : A total of 22 COI gene samples were amplified. The evolutionary tree constructed by 18 samples was consistent with the morphological identification results and 4 samples were different：Suncus murinus should be Crocidura lasiura,infant rats of Rattus losea and Rattus tanezumi was re-identified as Rattus rattus,infant rats of Microtus fortis（sample number：NH-1）needs further identification. Conclusion DNA barcoding can effectively correct the errors of morphological identification,thus combining the two methods could improve the accuracy of rodent identification.

Objective:To investigate the expression of HER2 and its relationship with clinicopathological features in patients with urothelial carcinoma.Methods:Urothelial carcinoma specimens collected from January 2019 to June 2022 were used. The expression of HER2, cytokeratin 20 and cytokeratin 5/6 was examined using immunohistochemistry. The HER2 expression was assessed according to the clinical pathological expert consensus on HER2 testing in urothelial carcinoma in China. Cases with HER2 2+/3+ were classified as HER2 positive. The relationship between HER2 expression and clinicopathological and molecular features was analyzed.Results:Four hundred and twenty-nine urothelial carcinoma specimens were analyzed, including 166 cases of raclical resection and 263 cases of local tumor resection. The median patient-age was 69 years (range: 31-93 years). The male: female ratio was 2.9∶1.0. The positive rate of HER2 was 45.7%(196/429). The positive rate of HER2 in patients with local tumor resection was higher than that in patients with radical resection [51.7%(136/263) vs.36.1%(60/166), P<0.05]. In the upper urinary tract (renal pelvis/ureter) urothelial carcinomas, the positive rate of HER2 was 35.2% (37/105). In bladder urothelial carcinoma, the positive rate of HER2 was 49.1%(157/320), and higher than that in upper urinary tract urothelial carcinoma ( P<0.05). In high grade urothelial carcinoma, the positive rate of HER2 was 52.8%(168/318) and higher than that in low grade urothelial carcinomas (25.2%, 28/111, P<0.01). In 166 radical resection specimens, the positive rate of HER2 was not differentially distributed by tumor pT stage [Ta (26.1%, 6/23), T1 (41.7%, 20/48), T2 (40.0%, 10/25), T3 (28.1%, 16/57), T4 (8/13) ( P>0.05)]. In urothelial carcinomas with muscle invasion, the HER2 positive rate was 35.8%(34/95), while the rate in non-muscle-invasion urothelial carcinoma was 36.6%(26/71, P>0.05). CK20 and CK5/6 were used to refine the urothelial carcinoma molecular subtypes. The positive rate of HER2 was highest in CK20 +/CK5/6 -group (124/194, 63.9%), followed by CK20 +/CK5/6 +group (18/40, 45.0%), CK20 -/CK5/6 -group (14/41, 34.1%) and CK20 -/CK5/6 +group (25/131, 19.1%, P<0.01). The positive rate of HER2 in micropapillary urothelial carcinoma was highest (14/15), followed by urothelial carcinoma with glandular differentiation (11/14), conventional urothelial carcinoma (161/360, 44.7%) and urothelial carcinoma with squamous differentiation (6/35, 17.1%, P<0.01). In the cases with lymph node metastasis, the positive rate of HER2 was 45.5% (10/22) and higher than the cases without lymph node metastasis (31.0%, 13/42). But there was no statistically significant association between HER2 expression and lymph node metastasis ( P>0.05). Conclusions:Expression of HER2 in urothelial carcinoma is closely correlated with tumor location, grade, histologic subtypes, molecular subtypes and surgical approach, but not with pT stage, muscle invasiveness or lymph node metastasis.

Objective To explore the CT and MRI imaging and clinicopathological features of extranodal NK/T cell lymphoma (NK/TCL). Methods Sixty-six patients with NK/TCL diagnosed from 2002 June to 2016 April in Beijing Tongren Hospital with intact CT and/or MRI imaging results were enrolled in this study. All the patients had tailed clinical information and follow-up. The imaging and clinicopathological features were analyzed retrospectively and their prognostic value on overall survival was analyzed. Results There were 49 males and 17 females with median age of 42 years. The median follow-up time was 18 months. The cases showed surrounding invasions including 10 cases (15.2 %) in soft palate, 5 cases (7.6 %) in hard palate, 2 cases(3.0 %) in tonsil, 8 cases(12.1 %) in upper lip, 13 cases(19.7 %) in maxillofacial soft tissue, 9 cases (13.6 %) in eyelid, 10 cases (15.2 %) in orbital, 3 cases (4.5 %) in maxilla, 6 cases (9.1 %) in pterygopalatine fossa,6 cases(9.1 %)in infratemporal fossa,3 cases(4.5 %)in skull base, 3 cases(4.5 %) in eyeball and 2 cases (3.0 %) in brain tissue. Kaplan-Meier survival analysis found that the 2-year overall survival rates of the patients with the involvement of hard palate, upper lip, maxillofacial soft tissue, eyelid, orbital, maxillary, eyeball and brain organizer were lower than those of the patients without the involvement of these sites(χ2values were 4.470,4.041,4.456,13.933,8.986,4.000,44.121,6.527,16.822,respectively, all P< 0.05). Further multivariate Cox regression analysis showed that maxilla and brain involvement were independent adverse factors (RR=34.717, 95 % CI 3.404-354.035, P=0.003; RR=37.545, 95 % CI 3.188-442.187, P= 0.004). Conclusions MRI and CT examinations are of great value in diagnosis and prognostic assessment of NK/TCL. Clinicians can make correct and timely diagnosis by comprehensive clinical, radiological and pathological features and can make a detailed clinical assessment to give patients appropriate treatment,thus improving the outcome of the NK/TCL patients.

Objective:To verify the value of whole genomic copy number variation (WGCNV) detection and scoring system in the diagnosis and prognosis of lung adenocarcinoma.Methods:Seventy-six lung adenocarcinoma specimens including ninety-one tumor samples and twenty adjacent non-tumor lung tissue samples were collected using Laser capture microdissection (LCM). Whole genomic amplification (WGA) was used to enrich DNA and construct a sequencing library for next generation sequencing (NGS). Changes of larger than 5Mb CNV in this study were analyzed and scored. The nuclear grading and score of tumor cells in the surgery and pleural effusion cytology of lung adenocarcinoma specimens were evaluated separately. For each case, we evaluated (1) nuclear size, (2) mitotic counts, (3) nuclear atypia, (4) atypical mitoses. The data of disease-free survive (DFS) and overall survive (OS) were collected for assessing the prognostic value of WGCNV score. Meanwhile, receiver operating characteristic (ROC) and area under curve (AUC) were used to define a cut-off value and evaluate the diagnostic significance in lung adenocarcinoma.Results:The WGCNV scores of twenty adjacent non-tumor lung tissue samples were treated as normal control and all of WGCNV scores of tumor samples range from 0 to 9.95, the median score was 2.7. The WGCNV scores were divided into three groups: low score group <1.74, medium score grade 1.74~4.23, high score grade >4.23. The WGCNV score was positively associated with the nuclear grade scoring ( r=0.780 90, P<0.001). The result for evaluation of prognostic value of the WGCNV scores showed that comparing with low WGCNV score group, Hazard Ratio (HR) of medium score group was 4.11 (95% CI=0.72~23.57) and high score group was 2.07 (95% CI=0.30~14.12). These results suggested that the risks of the medium and high WGCNV score group elevated. According to the analysis results of ROC curve, when the cut off value was 0.01, the sensitivity and specificity for lung adenocarcinoma diagnosis were 97.8% and 95.0% respectively, the positive predictive value (PPV) and negative predictive value (NPV) were 99.0% and 90.1%, respectively, the AUC was 0.981. In the differentiation of adenocarcinoma in situ (AIS) and minimally invasive adenocarcinoma (MIA) group and invasive adenocarcinoma group, when the cut off value was 1.8, the sensitivity and specificity between the two groups were 78.1% and 94.4%, and the PPV and NPV were 98.0% and 52.0%, respectively, the AUC was 0.896. Conclusion:This study verifies that WGCNV scoring system has a potential diagnostic and prognostic value in lung adenocarcinoma.

Objective:To verify the value of whole genomic copy number variation (WGCNV) detection and scoring system in the diagnosis and prognosis of lung adenocarcinoma.Methods:Seventy-six lung adenocarcinoma specimens including ninety-one tumor samples and twenty adjacent non-tumor lung tissue samples were collected using Laser capture microdissection (LCM). Whole genomic amplification (WGA) was used to enrich DNA and construct a sequencing library for next generation sequencing (NGS). Changes of larger than 5Mb CNV in this study were analyzed and scored. The nuclear grading and score of tumor cells in the surgery and pleural effusion cytology of lung adenocarcinoma specimens were evaluated separately. For each case, we evaluated (1) nuclear size, (2) mitotic counts, (3) nuclear atypia, (4) atypical mitoses. The data of disease-free survive (DFS) and overall survive (OS) were collected for assessing the prognostic value of WGCNV score. Meanwhile, receiver operating characteristic (ROC) and area under curve (AUC) were used to define a cut-off value and evaluate the diagnostic significance in lung adenocarcinoma.Results:The WGCNV scores of twenty adjacent non-tumor lung tissue samples were treated as normal control and all of WGCNV scores of tumor samples range from 0 to 9.95, the median score was 2.7. The WGCNV scores were divided into three groups: low score group <1.74, medium score grade 1.74~4.23, high score grade >4.23. The WGCNV score was positively associated with the nuclear grade scoring ( r=0.780 90, P<0.001). The result for evaluation of prognostic value of the WGCNV scores showed that comparing with low WGCNV score group, Hazard Ratio (HR) of medium score group was 4.11 (95% CI=0.72~23.57) and high score group was 2.07 (95% CI=0.30~14.12). These results suggested that the risks of the medium and high WGCNV score group elevated. According to the analysis results of ROC curve, when the cut off value was 0.01, the sensitivity and specificity for lung adenocarcinoma diagnosis were 97.8% and 95.0% respectively, the positive predictive value (PPV) and negative predictive value (NPV) were 99.0% and 90.1%, respectively, the AUC was 0.981. In the differentiation of adenocarcinoma in situ (AIS) and minimally invasive adenocarcinoma (MIA) group and invasive adenocarcinoma group, when the cut off value was 1.8, the sensitivity and specificity between the two groups were 78.1% and 94.4%, and the PPV and NPV were 98.0% and 52.0%, respectively, the AUC was 0.896. Conclusion:This study verifies that WGCNV scoring system has a potential diagnostic and prognostic value in lung adenocarcinoma.

The theory of “diagnosing diseases with sinews” means that through the diagnosis and examination of the channel sinews at the site of the lesion， the surplus and deficit state of qi and blood in the channel sinews， vessles and channels， and the degree of damage to the organism caused by the disease and evils， can be determined， forming the three elements （the nature of the disease， the location of the disease， and the disease tendency） of the disease diagnosis can be closely integrated to form the trinity of diagnostic modes， which is “examining the disease nature by sinews， identifying the pattern by sinews， and determining the tendency by sinews”. For intractable facial paralysis， the method of “diagnosing diseases with sinews” can be adopted， in which the morphological changes of the channel sinews are judged through diagnosis by observation， the traditional Chinese patterns are identified through diagnosis by palpation， and the points of meridian tendons and the circulation of tendon and treatment lines are determined through diagnosis by circulation. The “diagnosing diseases with sinews” not only helps to accurately determine the disease condition， patterns and development trend， but also helps to adopt targeted treatment for the disease and prevent the disease from spreading， and providing ideas and methods for the clinical diagnosis and treatment of intractable facial paralysis.