Objective To produce human papillomavirus type 6(HPV-6)virus-like particles with Escherichia coli expression system and study its immunogenicity.Methods HPV-6 L1 gene was inserted into pmkaryotic expression vector pTO-T7 and then expressed in Escherichia coli ER2566.The HPV-6 L1 protein was purified by ammonium sulfate precipitation,ion-exchange chromatography,and hydrophobic interaction chromatography.Then the purified HPV-6 L1 self-assembled into virus-like particle after removing 1,4dithiothreitol(DTr).The morphology of the virus-like particles was investigated with dynamic light scatter and transmission electron microscopy,and the immunogenicity was determined with in vitro pseudownons neutralization as8ay.Results HPV-6 L1 was expressed in soluble form in Escherichia coli.Following the removal of DTT,purified HPV-6 L1 protein could assemble into virus-like particles as 25 am in the radius.And the animal immunization test showed HPV-6 virus-like particles can elite hish titer neutralizing antibodies.Conclusion The bacterially expressed HPV-6 L1 VLP is highly immunogenieity and easy to produce.And it can be good candidate of HPV-6 vaccine.

Objectives To identify ABO blood type from a child having discrepant results in forward and reverse ABO blood grouping by serological identification and genetic testing.Methods After routine serological detection with ABO blood group,the ABO gene and ABO blood group-A subgroup genotype were tested by PCR-SSP method.Results The Serological results showed that the specimen was positive type A (anti-A:1 +w),the reverse type is O type (Ac:2 +,Bc:4 +);PCRSSP A-subtype typing showed that the genotype of the child was Ax14/O2.Conclusion Difficult blood type identification sometimes need to combine serology and molecular biology methods to confirm.In this case,the phenotype of the child was Ax,and the genotype was Ax14/O2.

Objective To evaluate percutaneous AngioJet thrombectomy in the treatment of iliofemoral deep venous thrombosis (DVT).Methods A total of 36 patients with 38 legs of lower extremity DVT treated by AngioJet mechanical thrombectomy from Feb 2016 to Dec 2016 were analyzed retrospectively.The effect of lower limb venous recanalization was evaluated by observing the intraoperativeangiography and the results of postoperative follow-up including complaints,signs,and lower extremity venous ultrasound or CT and Villalta scores.Results Thrombosis was completely dissolved by AngioJet thrombectomy device in 32 out of 38 legs (84.2％) at first stage.Slight bleeding occurred in 4 cases,1 patient could not tolerate the operation.23 patients were followed-up for 6 months,venous patency was present in 21 of 23 patients (91.3％).11 patients complete the one-year follow-up,9 patients (81.8％) were in the 0-4 Villalta group.Conclusions AngioJet can effectively and safely remove thrombus in the lower extremity deep venous system.It is especially advantageous for patients who have contraindications for thrombolysis with a satisfactory short term venous patency.

Objective:To study the effect of resveratrol (RES) on interleukin-1β (IL-1β)-induced chondrocytes and its pathways of action.Methods:Wistar mammary rat chondrocytes were extracted and divided into 5 groups: control group, IL-1β group, RES+IL-1β group, RES+IL-1β+EX-527 [silent information regulator 1 (SIRT1) inhibitor] group and RES+IL-1β+AS [frame transcription factor O1 (FOXO1) inhibitor] group. Quantitative real time polymerase chain reaction (qRT-PCR) was used to detect SIRT1, forkhead FOXO1 and matrix metalloproteinase 3 (MMP-3) mRNA expression. Protein expression of chondrocyte type Ⅱ collagen (Col-Ⅱ) detected by immunofluorescence, and the expression of chondrocyte SIRT1 and p-FOXO1/FOXO1 was measured by Western blot. The expression of chondrocyte inflammatory factors IL-6 and TNF-α was measured by enzyme-linked immunosorbent assay. One-way analysis of variance (ANOVA) was performed and two-way comparisons between groups were made using the least significant difference (LSD) method. P< 0.001 was statistically significant. Results:Compared to normal chondrocytes, the mRNA and protein expressions of Col-Ⅱ, SIRT1, FOXO1 and p-FOXO1/FOXO1 in chondrocytes induced by IL-1β was significantly decreased ( P<0.001). The secretion of tumor necrosis factor (TNF)-α [(24.70±2.84), t=19.24, P<0.001] and IL-6 [(3.35±0.28), t=12.97, P<0.001] was significantly increased, and the mRNA expression of MMP-3 [(2.46± 0.23), t=12.61, P<0.001] was significantly increased. The mRNA and protein expressions of Col-Ⅱ, SIRT1, FOXO1 and p-FOXO1/FOXO1 were significantly increased. The secretion of TNF-α [(12.60±1.05), t=10.14, P<0.001] and IL-6 [(2.00±0.15), t=9.89, P<0.001] was significantly reduced by RES treated IL-1β-induced chondrocytes. mRNA expression of MMP-3 [(1.30±0.14), t=10.460, P<0.001] was decreased. After adding SIRT1 inhibitor EX-527 or FOXO1 inhibitor AS, RES significantly reduced the mRNA and protein expression of Col-Ⅱ, SIRT1, FOXO1 and p-FOXO1/FOXO1 in IL-1β-induced chondrocytes ( P<0.001). The secretion of TNFα and IL-6 was significantly decreased ( P<0.001), and the mRNA expression of MMP-3 was significantly decreased ( P<0.001). Conclusion:RES significantly ameliorates IL-1β-induced cartilage extracellular matrix egradation and inflammatory responses via the SIRT1/FOXO1 pathway.

Immune imbalance is believed to play a dominant role in the pathogenesis of chronic urticaria.While Th1/Th2 imbalance used to be considered as the main contributing factor of the development of chronic urticaria.Recently,Th17/Treg imbalance is found to be an important immune mechanism leading to the development of chronic urticaria.To be more specific,according to traditional Chinese medicine(TCM)'s comprehensive understanding of the etiology of chronic urticaria.it is generally believed that the pathogenesis of chronic urticaria is due to a lack of innate endowment,a lack of solidity of the body surface,and repeated exposure to six pathogenic factors.Another possible reason lies with dietary disorders that generate heat and wind or chronic illness and weakness and loss of nourishment of qi and blood.Therefore,in terms of the treatment,from the perspective of sthenia syndrome,it is advisable to remove the wind and disperse the pathogenic factors and clear the dampness and heat.From the perspective of asthenia syndrome,it is advisable to nourish the qi and blood and support the righteousness.As for mixed excess and deficiency,both support the healthy atmosphere and dispel the pathogenic factors are important.Regarding the effects of TCM on the balance of Th17/Treg in chronic urticaria and immune diseases,it mainly involved herbal compounding,herbal active ingredients and single herbs.However,the research attention has been drawn to investigating the role of TCM in the treatment of chronic urticaria and various immune diseases based on the research outcomes in modern pharmacological research.This can not only provide scientific evidence for TCM treatment of chronic urticaria,but also bring benefits to more patients with immune diseases.Therefore,the author reviews the recent research progress of TCM on the effects of Th17/Treg immune imbalance in chronic urticaria and other immune diseases by explaining the effects of Th17 and Treg cells in chronic urticaria.

In the differentiation and treatment of recurrent urinary tract infection （rUTI） from the perspective of the seminal orifice， it is proposed that the urinary tract belongs to the category of "seminal orifice"， and the physiological process of urination is closely related to the function of the seminal orifice. From the three dimensions of orifice body， orifice pivot and orifice spirit， the physiological relationship between seminal orifice and the function of five zang-organs （脏） is constructed， that is， lung heat， yin damage and pathogen counter-restriction lead to malnutrition of orifice body； burning heart fire and spirit disorder lead to unfavorable orifice spirit， and kidney deficiency， liver constraint and spleen stagnation lead to unfavorable orifice pivot. In the early stage of rUTI， there is usually unfavo-rable orifice pivot， for which the treatment principle should be treating the root and the branch simultaneously， consi-dering both deficiency and excess， and paying attention to the management of accompanied symptoms. Zishui Qinggan Beverage （滋水清肝饮） and Modified Shenzhuo Decoction （肾着汤加减） are often selected based on syndrome differentiation. In the middle stage， lack of nourishment of the orifice body and unfavorable orifice spirit and pivot coexist， and the treatment should be draining the orifice and unblocking strangury， commonly withmodified Qingxin Lianzi Beverage （清心莲子饮）. In the late stage， loss of nourishment of the orifice body is the main pathogenesis， and it is necessary to further nourish the orifice body to prevent recurrence， and modifed Wuzi Yanzong Pills and Erxian Decoction （五子衍宗丸合二仙汤） is often used. Furthermore， the specific medicinals should be selected targeting at the orifice body， orifice pivot， and orifice spirit， so as to nourish orifice body by dispelling external pathogens and rectify healthy qi， to drain orifice pivot by freeing emotions and minds and unblocking qi movement， and to calm orifice spirit by unblocking heart and kidney and nourishing heart spirit.

Objective:To explore the relationship and underlying mechanism between exosomes derived from doxorubicin-resistant osteosarcoma cells and MDR1 and miRNAs. Methods:MG63 and U2OS cell lines were selected to construct doxorubicin-resistant strains, and the 50% inhibitory concentration (half maximal inhibitory concentration, IC 50) of drug-resistant and sensitive strains was detected by MTT, and fluorescence staining was performed at intervals of 15 min between 15 and 120 min to detect the change of fluorescence intensity. RT-PCR and Western Blot were used to detect the expression levels of MDR1 P-gp to verify the drug resistance of osteosarcoma cells. Exosomes were identified by particle size analysis and Western Bolt detection. The endocytosis of PKH26-labeled exosomes from doxorubicin-resistant cells was observed, and the proliferation level and migration of exosomes from doxorubicin-resistant cells co-cultured with osteosarcoma cells were detected by MTT assay and cell scratch assay. The differential expression levels of miRNAs in osteosarcoma-sensitive and drug-resistant cells were verified by sequencing and bioinformatics analysis and RT-PCR assay. Tumor growth, serum exosome identification and mRNA expression level of miR-21-5p in tumor-bearing nude mice between normal osteosarcoma cell group and drug-resistant group, drug-resistant+normal exosome group, drug-resistant+drug-resistant+drug-resistant exosome group were observed. MDR1 expression level in tumor tissue was detected by RT-PCR, Western Blot and immunohistochemistry. Results:The IC 50 of two adriamycin resistant strains were 2.21 vs. 11.81 μg/ml and 0.93 vs. 11.81 μg/ml, respectively, and the fluorescence intensity decreased faster than that of normal strains. The relative mRNA expression levels of MDR1 in two cell lines were normal 1.12±0.16, 1.02±0.11 and drug-resistant 2.15±0.10, 2.127±0.12, respectively. The relative protein expression of P-gp was normal 0.92±0.11, 0.73±0.10 and drug-resistant 0.46±0.03, 0.30±0.04, the differences were statistically significant ( P<0.05). Drug-resistant exosomes can enter osteosarcoma cells through endocytosis and concentrate in the cytoplasm when co-cultured with normal strains. Osteosarcoma cells were co-cultured with drug-resistant exosomes at 2, 4, 6, and 8 μg/ml adriamycin, respectively. Compared with normal group, the proliferation level in drug-resistant group was significantly increased. Compared with the normal cell group 35.95±3.92, 6.72±3.55 and the normal exosome group 51.22±5.55, 19.31±1.93, the drug-resistant cell group 54.20±9.32, 19.24±2.88 and drug-resistant exosome group 76.40±5.41, 30.26±4.87, all had significantly higher cell mobility, the difference was statistically significant ( P<0.05). Exosome sequencing and biogenic analysis of 10 highly upregated miRNAs to validate mRNA expression differences between normal and drug-resistant strains by RT-PCR, showing a significant increase in miR-21-5p expression level of drug-resistant strains (5.89±0.26 vs. 0.99±0.06; 1.05±0.07 vs. 8.80±0.93, P<0.05), the difference was statistically significant ( P<0.05). In MG63 and U2OS, the normal cell group and drug-resistant cell group, and the normal exosome group and drug-resistant exosome group were compared, the tumor volume and the terminal tumor weight of nude mice were increased to varying degrees. MRNA relative expression levels of miR-21-5p in serum exosomes of nude mice after drug intervention were 0.86±0.07 and 0.86±0.05 in normal cell group, respectively. The values were 1.13±0.12, 1.14±0.12 in drug-resistant cell group, 0.71±0.05, 0.75±0.03 in normal exosome group, and 0.90±0.07, 0.93±0.04 in drug-resistant exosome group. Compared with normal and drug-resistant strains, the expression levels of normal and drug-resistant exosome groups were increased, with statistical significance ( P<0.05). Conclusion:The exosomes of drug-resistant cells in osteosarcoma could enhance the proliferation level and migration ability of cells through intercellular transfer of MDR1 and miRNAs. The expression of MDR1 and miR-21-5p in drug-resistant cells and tumor-forming nude mouse serum and tumor tissues were up-regulated which suggested that it might be involved in regulating the drug resistance process of osteosarcoma.

OBJECTIVE: To analyze the efficacy of integrated minimally invasive surgery for iliac vein compression syndrome with varicose veins of lower extremities. METHODS: From January 2017 to January 2018, 11 patients with iliac vein compression syndrome accompanied by varicose veins of lower extremities underwent left iliac vein stent implantation and radiofrequency thermal ablation of lower extremity veins in the First Affiliated Hospital of Zhejiang University School of Medicine. The left iliac vein stent was implanted through the puncture point approach of the main great saphenous vein, and then radiofrequency thermal ablation of the main saphenous vein was performed. Rivaroxaban and aspirin were administered from the day of surgery for 6 months and 12 months, respectively. After discharge, patients were followed up for more than 6 months. The lower extremity veins, iliac veins were reexamined by Doppler ultrasound or CT angiography at 2 weeks, 2 months and 6 months after surgery. RESULTS: The operations were successfully performed in 11 patients, and no complication was observed during the operation. The rates of soreness and swelling remission, pigmentation and skin quality improvement, and the iliac vein stent patency were 100%. No varicose vein recurrence, iliofemoral vein thrombosis and pulmonary embolism were found. CONCLUSIONS Integrated minimally invasive surgery is safe, effective and less invasive for iliac vein compression syndrome with varicose veins of lower extremities.
Humans ; Lower Extremity ; surgery ; May-Thurner Syndrome ; complications ; surgery ; Minimally Invasive Surgical Procedures ; standards ; Radiofrequency Ablation ; Saphenous Vein ; surgery ; Treatment Outcome ; Varicose Veins ; complications ; surgery

Objective To determine the radiation dose of sensitive organs under different protective methods in lung CT scanning environment, and to explore the best protective scheme of corresponding organs. Methods Annealed thermoluminescence dose elements were placed in the stomach, liver, colon, and thyroid gland of a simulated human body model. The dose effect experiment of protective methods included non-protective group, half lead apron group, and full lead apron group. The dose effect experiment of protective thickness included 0.50 mmpb full lead apron group and 0.35 mmpb full lead apron group. The same exposure conditions of lung CT scan were used in the above experiments. Results Compared with the non-protective group, the exposure dose of the stomach, liver, colon, and thyroid gland increased significantly in the half lead apron group (P < 0.05), and the exposure dose of the thyroid gland and colon decreased significantly in the full lead apron group (P < 0.05). There were no significant differences in the exposure dose of the liver, stomach, and colon in the simulated human body model between the 0.35 mmpb full lead apron group and the 0.50 mmpb full lead apron group. Conclusion For lung CT scan, the protective measure of lead apron may not reduce the exposure dose of subjects. The protective thickness of lead apron does not necessarily have a substantial influence on the exposure dose of human body.

Objective    To investigate the preoperative localization of pulmonary glabrous nodules. Methods    A total of 192 patients admitted to General Hospital of  Northern Theater Command from April 2012 to September 2019 were selected for the study. There were 95 males and 97 females at an age of 56.47±11.79 years. All patients completed preoperative examination, and were divided into a positioning group (n=97) and a non-positioning group (n=95) according to whether the preoperative positioning was performed. And the surgical indicators between the two groups were compared. According to the substance of ground-glass opacity, they were divided into a pure ground-glass nodules group (n=23) and a mixed ground-glass nodules group (n=74) in the positioning group and a pure ground-glass nodules group (n=14) and a mixed ground-glass nodules group (n=81) in the non-positioning group . According to the size and distance of the nodules from the pleura and whether the nodules could be detected, the corresponding linear function was obtained. Results    The operative time of methylene blue localization group was shorter than that of the no localization group. In the scatter plot, the corresponding diameter and depth of the nodules and the corresponding coordinate points which can be explored were described. And linear regression was performed on all the coordinate points to obtain the linear function: depth=0.648×diameter–1.446 (mm). It can be used as an indication for the preoperative localization of pure ground-glass nodules in Da Vinci robotic surgery. Linear function: depth=0.559 5×diameter+0.56 (mm). It can be used as an indication of preoperative localization of mixed ground-glass nodules in Da Vinci robotic surgery. Conclusion    This equation can be used as a preoperative indication for clinical peripheral pulmonary ground-glass nodules.