Objective To research the effect of PEGylation on rBla g 2 from Blattella germanica.Methods rBla g 2 allergen expressed in E.coli was purified by Ni+affinity chromatographY,then was PEGylated by mPEG2-NHS(Mr,10×103).The PEG-rBla g 2 was purified by CM-Sepharose,a cation exchange chromatography. SDS-PAGE,Western blot and ELISA were used to characterize its biologicat actovoty.Results The relative molecular mass of the purified rBla g 2 was about 39×1023.PEG-rBla g 2 was analyzed by SDS-PAGE.Five bands were visualized by staining with Coomassie Brilliant Blue R-250,while seven bands by staining with I2-KI.Cation exchange chromatography could separate rBla g 2 and PEG-rBla g 2.The 100×103(Mr)and 130×103(Mr)of PEG-rBla g 2 could combined with the special IgE from sera of one cockroach-allergic patient by Western blot.The immunological activities of PEG-rBla g 2 in vitro decreased remarkablv bv ELISA,which was only 42%of rBla g 2.Conclusion PEGylated allergen can retain the ability of combining with special IgE from sera,while its immunological activities reduce enormously,which establishes the basic work of researching recombinant low-sensitive allergens.

AIM: In this study, CD147 antibody was used to carry out targeted modification of nanoparticles for protein kinase Cε (PKCε)-siRNA gene therapy to target lung cancer cells.The inhibitory effects of the nanoparticles on the proliferation and invasion of the lung cancer cells were observed.METHODS: The magnetic nanoparticles targeting CD147 protein were assembled as gene vector.The expression of CD147 in the lung cancer cells was observed under laser scanning confocal microscope.The cells were divided into CP group, CN group and LP group as the experimental groups. Targeted nanoparticles were used as CA group.Non-transfected cells were used as control group.The cell transfection was carried out with 250 ng plasmids/well in 6-well plate.The effect of nanocontrast agent on the cell endocytosis was observed under laser scanning confocal microscope.The mRNA expression of PKCε was detected by RT-qPCR.The protein expres-sion of Ki67, MMP3, PKCε, Wnt1 and GAPDH was determined by Western blot.The cell proliferation ability was detec-ted with colony formation assay.The cell invasion ability was detected by Transwell method.RESULTS: The expression of
CD147 protein in the human lung cancer A549 cells was confirmed by immunofluorescence staining.The endocytosis of siRNA into the A549 cells in CP group was observed with the highest efficiency as compared with CN group and LP group. The relative mRNA expression of PKCε in the A549 cells of CP group, CN group, LP group and CA group were (9.76 ± 0.18)%, (98.51 ±0.32)%, (99.17 ±0.16)% and (99.68 ±0.11)%, respectively.The difference between CP group and control group was statistically significant (P <0.05).No significant difference among CN group, LP group and control group was observed.The protein expression of PKCε, Ki-67, MMP3 and Wnt1 in CP group was significantly reduced, and the protein expression levels among CN group, LP group and control group had no significant difference.The colony number in CP group was significantly smaller than that in control group (P <0.05).The effective colony numbers in CN group, LP group and CA group had no significant difference as compared with control group.The number of the invading cells in CP group was significantly less than that in control group (P <0.05).The numbers of the invading cells in CN group, LP group and CA group had no significant difference as compared with control group.CONCLUSION: Nanogene vector targe-ting CD147 can carry PKCε-siRNA to conduct gene therapy efficiently on the lung cancer cells to achieve effective inhibito-ry effects on the proliferation and invasion of the lung cancer cells.

Aim To investigate the effects of dalbinol on proliferation and apoptosis of human colon cancer HCT1 16 cells and its mechanisms.Methods Anti-proliferative effect of dalbinol was evaluated by MTT assay.The morphological changes of apoptosis were observed by Hoechst33342 staining.Apoptotic rate and ROS generation were analyzed by flow cytometry.The related proteins of Wnt/β-catenin pathway and the ap-optosis-associated proteins expression were measured by Western blot.Results The growth of HCT1 16 treated with dalbinol was inhibited in a dose and time dependent manner with IC50 (4.8 ±0.53 ),(2.5 ± 0.43)and (0.6 ±0.22)μmol·L-1 at 24,48 and 72 h,respectively.Typical morphological changes of ap-optosis such as cell shrinkage,karyopyknosis and nu-clear condensation were observed by Hoechst33342 staining.Meanwhile,the apoptotic rate and intracellu-lar ROS generation of dalbinol were both increased dose-dependently. Western blot results showed that dalbinol could activate the expression of cleaved Caspase-3 and cleaved PARP by decreasing anti-apop-totic protein levels such as Bcl-2 and Mcl-1 and in-creasing pro-apoptotic protein levels such as Bax and Bim,which induced further apoptosis.Moreover,dal-binol can reduce the protein expression of the total and nuclear β-catenin,but not cytoplasmic β-catenin by suppressing the protein expression of Dvl-2 and GSK-3β(pS9 ),as well as its target proteins c-Myc and Sur-vivin.Conclusion dalbinol can induce apoptosis in colon cancer HCT1 16 cells by upregulating the intra-cellular ROS generation and suppressing Dvl/GSK-3β/β-catenin pathway.

Objective To study the characteristics of the diseases spectrum in hospitalized children.Methods The items of main diagnosis,gender,age,etc.in the inpatients medical records home page data aged 0? 12 years old in second-level and above hospitals of Chongqing City during 2015 were performed the disease spectrum sequence and constituent ratio analysis for deeply digging the relation between the disease change trend with the age and sex.Results The top 5 main system diseases affecting children's health were respiratory system diseases (male 43.84％,female 45.74％),some conditions originated from prenatal period (male 12.01 ％,female 13.87 ％),infectious diseases and parasites diseases (male 9.48 ％,female 9.88 ％),digestive system diseases (male 9.09 ％,female 6.95 ％),injury,poisoning and some other consequences of external causes(male 6.86 ％,female 5.27 ％).Respiratory system diseases always ranked the first place,the sexual difference among various system disease was unobvious,the sequence of injury,poisoning and some other consequence of external causes was increased with the age increasing;in the ranking of respiratory system diseases,pneumonia(38.73 ％),acute bronchitis(17.29 ％) and acute tonsillitis(13.83％) were the top 3 diseases affecting children's health.Conclusion Respiratory system diseases are the concerned key point for children health promotion.The health and epidemic prevention departments should provide effective methods and measures for preventing respiratory system diseases in children.Furthermore,the targeted prevention and treatment measures should be proposed aiming at the difference of disease sequence in different age groups.

Objective To evaluate the effect of preoperative transarterial chemoembolization (TACE) for resectable hepatocellular carcinoma (HCC).Methods HCC patients undergoing up-front hepatectomy (group A) were compared with those receiving TACE before hepatectomy (group B).Results Tumor size decreased significantly after TACE (t =3.3 1,P =0.021).The rates of tumor encapsulation and liver adhesions were significantly more often seen in group B.There were fewer tumor-residual and more frequent necrosis in group B.Operative time in group B was longer (t =2.71 ,P =0.046).The average blood loss and complication rate were of no difference between the two groups.The occurrence of pleural effusion and intrahepatic recurrence rate in group A was higher than group B (x2 =3.85 ,P =0.031) (x2 =2.76,P =0.046).The overall survival rate from the second year postoperative in group B was higher than group A (x2 =3.37, P =0.043).Conclusions TACE could diminish tumor, advance encapsulation and reduce tumor-residual.Preoperative TACE does not improve 1-, 2-, and 3-year tumor-free survival rates but improve 1-, 2-, and 3-year overall survival rates.

Objective To evaluate the short-term and long-term elinical effect of hepatectomy using Tissue-Link & Cusa,compared to the Pringle maneuver.Methods Clinical data of 87 HCC patients who had received hepatectomy by the Pringle's Maneuver (group A) or Tissue-Link & Cusa (group B) were retrospectively analyzed.Results The average amount of bleeding in Group A was more than group B (t =2.030,P =0.023).The time of operation in group A was shorter than group B (t =-2.896,P =0.006).The postoperative supplement of albumin in group A was more than group B,the level of serum total bilirubin on 7th day after operation was higher than group B (P ＜ 0.05).There was no significant difference in postoperation complications and the time in hospital (P ＞0.05).The rate of incisional recurrence and the rate of metastasis in or out of the liver in group A were higher than group B (P =0.029,0.021,0.016).The 2-and 3-year tumor-free survival rates and the 3-year overall survival rates in group A were lower than that in group B (P =0.047,0.036,0.042).Conclusions Hepatectomy using Tissue-Link & Cusa is superior to the Pringle's maneuver for the treatment of primary hepatocellular carcinoma.It has a clear operative field,less operative bleeding,less damage to liver function and a lower relapse rate of incisional margin.

Objective To investigate the value of fluorescence imaging of labeling the hepatocellular carcinoma (HCC) cells with luciferase in tracking the pulmonary metastasis of HCC of nude mouse . Methods Low metastatic potential HCC97L and high metastatic potential HCC97H cells were transfected into HCC cell lines HCC97L-FL, HCC97H-FL with stable expression of luciferase. A total of 24 nude mice were randomly divided into HCC97L-FL group and HCC97H-FL group by the random number table method. Cell suspension was prepared by adding HCC97L-FL, HCC97H-FL cells (5×106) into the phosphate buffered saline (PBS, 50 μl) respectively. Cell suspension was injected into the caudal vein of nude mice. The model of tracking pulmonary metastasis of HCC with fluorescein was established. The gross and tissue samples of the pulmonary metastasis tracked by fluorescence imaging were observed.The data between two groups were compared using t test. Results Through fluorescence detection, intensive fluorescence could be observed in the HCC97L-FL, HCC97H-FL cells. After 2 to 3 weeks of modeling, fluorescence signal was collected from the nude mice lung in HCC97L-FL group. The fluorescence intensity enhanced gradually in 3-4 week. No fluorescence signal was observed in the nude mice lung in HCC97H-FL group. After 4 weeks of modeling, the fluorescence intensity was (1.73 ± 0.16) ×104 a.u. in HCC97H-FL group, which was significantly higher than that in HCC97L-FL group [(0.03 ±0.01) ×104 a.u.] (t=23.652, P<0.05). The nude mice lung in HCC97H-FL group was densely covered with metastatic tumors of varying sizes after 4 weeks of modeling. Some of the bigger tumors were visible on the lung surface. No metastatic tumor was observed in the HCC97L-FL group. The metastatic tumor was observed with an obvious cell nuclear atypia and characteristic feature of HCC. Conclusions Fluorescence imaging of using luciferase to label HCC cells can dynamically track the pulmonary metastasis of HCC of nude mouse . This technique provides a reliable quantitative research method for studying the growth, invasion and metastasis of HCC cells .

Purpose/Significance To address the challenge of low willingness to share scientific data among stakeholders in the tradi-tional Chinese medicine(TCM)industry,and to promote standardization,aggregation,sharing,and application of scientific data in the field of TCM.Method/Process By adhering to national standards for scientific data submission and management,integrating technologies such as blockchain and digital watermarking,the study aims to establish atrusted process for the exchange of scientific data in the field of TCM and build a TCM scientific data management platform.Result/Conclusion This platform will provide information technology support for regional TCM scientific data exchange,and effectively improve the efficiency and willingness of scientific data exchange within the region.

Objective To sort out the development history of disease control informatization in Hubei, and provide reference for the establishment of the ^"Hubei model^" of disease control informatization. Methods To understand the development process through data access and site investigation. Result Hubei CDC has built a provincial and municipal two-level data center; built a full-coverage security network; built a full traceable immune planning system; built a provincial, city, and county three-level video conference system; established a professional disease Control the information team. Conclusion The ^"Hubei model^" of disease control informationization has a solid foundation. It will adhere to the development concept of people as the core, business as the problem-oriented, big data as the means, multi-point trigger as the benchmark, and network security as the foundation. Play the role of technical support and information leadership in the control business.

Objective:To analyze the different clinicopathological features of intrahepatic cholangiocarcinoma with and without viral hepatitis.Methods:The clinicopathological data of 79 intrahepatic cholangiocarcinoma cases from Mar 2012 to Sep 2018 at Henan Provincial People's Hospital were retrospectively analyzed.Results:Twenty-five of the 79 patients with intrahepatic cholangiocarcinoma were accompanied by viral hepatitis. Those with viral hepatitis had a lower mean age at onset than those without [(53±11) years vs. (60±11) years, P=0.011], higher proportion of male patients (80% vs. 52%, P=0.017), higher AFP positive rate (40% vs. 19%, P=0.041), lower CA19-9 positive rate (48% vs. 72%, P=0.036), tend to occur in the right liver lobe (76% vs. 44%, P=0.009), a lower rate of bile duct invasion (16% vs. 41%, P=0.03), and were more likely to be mass type (mass type proportion 96% vs. 72%, P=0.032). Conclusions:Viral hepatitis is common in intrahepatic cholangiocarcinoma. Intrahepatic cholangiocarcinoma with and without viral hepatitis differ in clinicopathology. Intrahepatic cholangiocarcinoma with viral hepatitis is more likely to have the characteristics of hepatocellular carcinoma, while intrahepatic cholangiocarcinoma without viral hepatitis is more likely to have the characteristics of cholangiocarcinoma.