Design of hepadnavirus core protein-based chimeric virus-like particles carrying epitopes from respiratory syncytial virus.

Respiratory syncytial virus (RSV) is one of the most important pathogens causing respiratory tract infection in humans, especially in infants and the elderly. The identification and structural resolution of the potent neutralizing epitopes on RSV fusion (F) protein enable an "epitope-focused" vaccine design. However, the display of RSV F epitope II on the surface of the widely-used human hepatitis B virus core antigen (HBcAg) has failed to induce neutralizing antibody response in mice. Here, we used the hepadnavirus core protein (HcAg) from different mammalian hosts as scaffolds to construct chimeric virus-like particles (VLPs) presenting the RSV F epitope II. Mouse immunization showed that different HcAg-based chimeric VLPs elicited significantly different neutralizing antibody responses, among which the HcAg derived from roundleaf bat (RBHcAg) is the most immunogenic. Furthermore, RBHcAg was used as the scaffold platform to present multiple RSV F epitopes, and the immunogenicity was further improved in comparison to that displaying a single epitope II. The designed RBHcAg-based multiple-epitope-presenting VLP formulated with MF59-like adjuvant elicited a potent and balanced Th1/Th2 immune response, and offered substantial protection in mice against the challenge of live RSV A2 virus. The designed chimeric VLPs may serve as the potential starting point for developing epitope-focused vaccines against RSV. Our study also demonstrated that RBHcAg is an effective VLP carrier for presenting foreign epitopes, providing a promising platform for epitope-focused vaccine design.

Impact of bariatric surgery on glucose and lipid metabolism and liver and kidney function in food-induced obese diabetic rats.

BACKGROUND: Obesity usually causes diabetes mellitus (DM) and is a serious danger to human health. Type 2 DM (T2DM) mostly occurs along with obesity. Foodborne obesity-induced DM is caused by an excessive long-term diet and surplus energy. Bariatric surgery can improve the symptoms of T2DM in some obese patients. But different types of bariatric surgery may have different effects. AIM: To investigate the effect of bariatric surgery on glucose and lipid metabolism and liver and kidney function in rats. METHODS: Male Sprague-Dawley rats aged 6-8 wk underwent Roux-en-Y gastric bypass surgery (RYGB), sleeve gastrectomy (SG), or gastric banding (GB). Glucose and insulin tolerance tests, analyses of biochemical parameters, histological examination, western blot, and quantitative real-time polymerase chain reaction were conducted. RESULTS: In comparison to the sham operation group, the RYGB, SG, and GB groups had decreased body weight and food intake, reduced glucose intolerance and insulin insensitivity, downregulated biochemical parameters, alleviated morphological changes in the liver and kidneys, and decreased levels of protein kinase C ß/ P66shc. The effect in the RYGB group was better than that in the SG and GB groups. CONCLUSION: These results suggest that RYGB, SG and GB may be helpful for the treatment of foodborne obesity-induced DM.

Concomitant gallstone disease was not associated with long-term outcomes in ursodeoxycholic acid-treated patients with primary biliary cholangitis.

OBJECTIVES: Primary biliary cholangitis (PBC) is a rare disease characterized by intrahepatic cholestasis, whereas gallstone disease (GD) is common. In this study, we aimed to investigate the prevalence and impact of GD on the prognosis of PBC in China. METHODS: Medical records of the PBC patients were retrospectively reviewed and their follow-up data were obtained via regular structured, standardized telephone interviews. GD was defined as gallstones on ultrasonography or a history of cholecystectomy for gallstones. Propensity score matching (PSM) and Cox regression analysis were performed. The primary end-point was liver-related death and/or liver transplantation. RESULTS: A total of 985 ursodeoxycholic acid (UDCA)-treated PBC patients were enrolled with a median follow-up duration of 5.3 years (range 1.0-20.9 years). Among them, 258 (26.2%) had GD, including 157 (22.9%) of non-cirrhotic and 101 (33.8%) of cirrhotic patients. Compared with PBC without GD, those with GD were older, more often had type 2 diabetes mellitus, and had a more severe liver disease at baseline. After PSM (1:2), 229 PBC patients with GD were matched with 458 PBC patients without GD based on age, sex, cirrhosis, and total bilirubin level. The transplant-free survival and incidence of hepatic events were similar between the two groups. Furthermore, multivariate Cox regression analysis showed that concomitant GD was not independently associated with a worse prognosis for PBC patients. CONCLUSION: Concomitant GD was common but was not associated with long-term outcomes in patients with UDCA-treated PBC.

[Realization and Clinical Study of Robotic Technology in Cardiovascular Interventional Surgery].

The high incidence of cardiovascular diseases is a serious threat to human health, and endovascular surgery has become the standard treatment for most interventional cardiovascular diseases. The robotassisted endovascular surgery system further enhances surgeons' ability to perform minimally invasive endovascular procedures in interventional cardiology. This study presents a new robotic technique for coronary intervention from the perspective of clinical application. Aiming at clinical application scenarios, this scheme proposed an intuitive guide wire catheter mechanism design, which accurately and perfectly simulates the doctor's hand movements, realizes the positive and negative direction translation of the guide wire catheter, accurate torque control of the guide wire rotation and locking. The results of animal test showed that the R-OneTM has a high degree of dexterity, accuracy and stability,and meets the clinical needs.

Concomitant extrahepatic autoimmune diseases do not compromise the long-term outcomes of primary biliary cholangitis.

BACKGROUND: Primary biliary cholangitis (PBC) patients often have concomitant extrahepatic autoimmune (EHA) diseases including Sjögren's syndrome (SS), systemic sclerosis (SSc), rheumatoid arthritis (RA), and autoimmune thyroid disease. The present study aimed to describe the prevalence of EHA diseases in PBC and explore the impact of EHA diseases on the long-term outcomes of PBC in Chinese patients. METHODS: Medical records of PBC patients diagnosed in our institute were retrospectively reviewed. Patients were followed up by a standardized telephone interview. The endpoints were defined as liver-related death and/or liver transplantation. RESULTS: Totally 247 of the 985 (25.1%) PBC patients enrolled in the study had at least one concomitant EHA disease. Sjögren's syndrome (n = 140, 14.2%) was the most frequent one, followed by rheumatoid arthritis (RA) (n = 56, 5.7%) and Hashimoto's thyroiditis (n = 45, 4.6%). Patients with EHA diseases were more common in females (P < 0.001) and in those with a family history of autoimmune disease (P = 0.017). Overall, no differences were found between PBC patients with and without EHA diseases in terms of biochemical response rates to ursodeoxycholic acid, the incidence of hepatic events, or transplant-free survival. RA and EHA ≥ 2 were protective factors for hepatic events in univariate Cox analysis, but the results became insignificant in multivariate analysis. CONCLUSIONS: Concomitant EHA diseases were common in PBC patients but did not compromise the long-term outcomes of PBC.

Optical Density Value and Ratio as Novel Indexes for Nanoemulsion Sterile Filtration Process Control or Characterization.

Sterile filtration is an effective method to remove any microorganisms present during nanoemulsion preparation. However, it lacks effective control parameters. Here, we established a simple and rapid approach for the process control of nanoemulsion sterile filtration by utilizing optical density detection as a process control parameter. During sterile filtration, the optical density or optical density ratio of the filtrate were continuously monitored to explore the correlation between optical density and the emulsion content and the change in the optical density ratio before and after sterile filtration. In the emulsion stability test, the optical density ratio was determined. A good correlation was obtained between the optical density and the nanoemulsion content during sterile filtration, thereby reducing sterile filtration loss. The optical density ratio changed significantly after sterile filtration, indicating that it could be used as a process control parameter to monitor leakage during emulsion sterile filtration. The optical density ratio can be a characterization index for stability monitoring as it is more sensitive than particle size detection and more convenient than large particle detection. These parameters may be used for sterile filtration process control and as an index for nanoemulsion characterization. This approach overcomes the limitations of existing nanoemulsion characterization methods.

[Risk factors of oral Helicobacter pylori infection among children in two kindergartens in Suzhou and the effects of oral cleaning on reducing oral Helicobacter pylori infection].

OBJECTIVE: This work aimed to identify the risk factors of Helicobacter pylori (H. pylori) infection in preschool children and provide effective measures for the prevention and reduction of the incidence of H. pylori infections. METHODS: A total of 204 children from two kindergartens in Suzhou city were recruited through a questionnaire survey. Risk factors were selected through the single factor paired data χ² test and multiple factor Logistic regression analysis. Oral and gastric H. pylori infections were detected by using H. pylori saliva detection (HPS) and ¹³C-urea breath test (¹³C-UBT). Special toothpaste for H. pylori control was selected for oral cleaning. Oral H. pylori infection rates at 2 months after special toothpaste treatment were examined by using HPS. RESULTS: The high-risk factors of H. pylori infections among preschool children included poor personal hygiene habits, such as the nibbling of fingers and the avoidance of hand-washing before meals, diet, and parent's history of stomach disease. Among the 204 subjects enrolled in this study, 158 (77.45%), 37 (18.14%), and 28 (13.73%) were HPS positive, ¹³C-UBT positive, and HPS and ¹³C-UBT positive, respectively. The incidence of oral H. pylori infections was significantly higher than that of gastric H. pylori infections (P<0.01). The positive rate of infections significantly decreased after special toothpaste treatment (P<0.01). This result indicates that the intervention was effective. CONCLUSIONS: Children must receive education on good eating habits. Individualized dining habits or the use of public chopsticks must be implemented. H. pylori infections must be detected as early as possible. Specific toothpaste for oral cleaning must be selected. These approaches could drastically prevent or reduce the incidence of H. pylori infections among preschool children.

The biological functions of LGR5 in promoting non-small cell lung cancer progression.

BACKGROUND: The Leucine-rich repeat-containing G-protein-coupled receptor 5 (LGR5), which is used as a marker of adult stem cells and colorectal cancer stem cells (CSCs), is closely associated with the progression of non-small cell lung cancer (NSCLC). This study aimed to identify the clinical significance and biological function of LGR5 in NSCLC. METHODS: Quantitative reverse transcription polymerase chain reaction (RT-PCR) was applied to detect the expression of LGR5 and stemness-related genes in 22 NSCLC patients, and the clinical significance of LGR5 in NSCLC progression was estimated by statistical analysis. LGR5 overexpressing A549- and H1299-transfected cells were established, and CCK-8 and clone formation assays were used to test the proliferation ability. A wound-healing assay was utilized to clarify the migration ability. The invasion ability was confirmed via the Transwell assay kit. RESULTS: LGR5 expression was markedly higher in NSCLC tissues than in the matched adjacent normal tissues, and had a trend to associate with tumor size, lymph node metastasis, and TNM stage. The proliferation rates, clone formation rates, wound healing rates, number of invasive cells, and the NOTCH1 expression of the LGR5 overexpressing groups, were significantly higher than those of the control groups. CONCLUSIONS: LGR5 plays an essential role in NSCLC tumorigenesis and is closely associated with the proliferation, metastasis, and invasion of NSCLC cells. LGR5 may promote NSCLC progression via NOTCH1 and could be a new target for gene-targeted therapies for NSCLC.

Complete plastome sequence of Manilkara zapota (L.) P.Royen (Sapotaceae).

iThe plastome of Manilkara zapota is found to be 158,386 bp long with the typical quadripartite structure of angiosperms, contains two inverted repeats (IRs) of 26,099 bp each, a large single-copy (LSC) region of 87,745 bp, and a small single-copy (SSC) region of 18,443 bp. The plastome contains 114 genes, consisting of 80 unique protein-coding genes, 30 unique tRNA genes, and 4 unique rRNA genes. The overall A/T content in the plastome of M. zapota is 63.00%. The phylogenetic analysis indicated that M. zapota is close to Sideroxylon wightianum within Sapotaceae in this study.

Clinical Significance of B7-H3 Expression During the Progression of Hepatitis B Virus Infection.

B7-H3, one of the costimulatory members participating in checkpoint pathway, has been shown to be upregulated after hepatitis B virus (HBV) infection. To further explore the clinical significance of dynamic B7-H3 expression during the progression of HBV infection, we systematically investigated the expression pattern of B7-H3 and the correlation of B7-H3 expression with the ratio of T lymphocyte subsets and clinical parameters at different stages in the course of the disease. Flow cytometry and enzyme-linked immunosorbent assay data showed that soluble form of B7-H3 (sB7-H3) was positively correlated with the frequency of Treg cells in acute hepatitis B (AHB), chronic hepatitis B (CHB), and hepatocellular carcinoma patients with HBV infection (HBV-HCC). Membrane form of B7-H3 (mB7-H3) expressed on Treg cells and monocytes was positively correlated with the frequency of Treg cells in CHB. SB7-H3 had relationship with mB7-H3 expressed on Treg cells and monocytes at different stages during HBV infection, except for HBV-HCC. MB7-H3 expressed on Treg cells was positively correlated with that on monocytes in AHB, CHB, HBV-liver cirrhosis, and HBV-HCC. The B7-H3 expression was positively correlated with aspartate aminotransferase and alanine aminotransferase levels in CHB and sB7-H3 level was higher in late tumor/node/metastasis (TNM) stage in HCC. Higher mB7-H3 expression was associated with greater tumor size, later TNM stage, and worse prognosis in HBV-HCC indicated by immunohistochemistry. Taken together, these results suggested that B7-H3 might contribute to the progression of HBV infection by triggering inhibitory signals in effector T cells and it was closely associated with the progression and poor prognosis during HBV infection. B7-H3 could be utilized as a potential clinical indicator and a potential target for therapeutic strategies against HBV infection.

The role of LGR5 and ALDH1A1 in non-small cell lung cancer: Cancer progression and prognosis.

The Leucine rich repeat containing G protein coupled receptor 5 (LGR5), may be a candidate marker of non-small cell lung cancer (NSCLC) cells with stem cell-like properties. Aldehyde dehydrogenase 1A1 (ALDH1A1) is one of NSCLC stem cell markers. To identify the relationship of LGR5 and ALDH1A1 in NSCLC, we analyzed the expression of LGR5 and ALDH1A1 in NSCLC samples, and determined their clinical significance. We performed quantitative RT-PCR for LGR5 and ALDH1A1 expression in 24 NSCLC patients, and showed that LGR5 and ALDH1A1 mRNA were frequently increased in NSCLC tissues in comparison to that in adjacent normal tissues (p = 0.0005 and p < 0.0001, respectively). Besides, the expression of LGR5 and ALDH1A1 mRNA has a significant correlation (r = 0.416, P = 0.0483). The expression of LGR5 and ALDH1A1 in 109 NSCLC tumors and 50 adjacent normal tissues were detected by immunohistochemistry. Positive LGR5 and ALDH1A1 expression was defined in 28.4% and 41.3% of the NSCLC tumors, respectively. Further analysis indicated that 24 of these LGR5⁺ (24/31) samples expressed ALDH1A1(r = 0.3883, p < 0.0001), we also found co-localization of LGR5 and ALDH1A1 in tumor tissue samples. LGR5 and ALDH1A1 expression was significantly associated with higher pathological TNM stage of the disease (stage I + II and III + IV) (P = 0.0311 and p = 0.0221, respectively), the co-expression of LGR5 and ALDH1A1 was associated with nodal status (p = 0.0424). High expression of LGR5 or ALDH1A1 was related to poor prognosis (P = 0.0125 and p = 0.0410, respectively), and NSCLC patients with co-expression of LGR5 and ALDH1A1 had a poorer prognosis than the others (P = 0.0011). Both of them can be an independent risk factor of a poorer prognosis (P = 0.016 and P = 0.024, respectively). The expression of LGR5 and ALDH1A1 were closely associated with the tumorigenicity, metastasis and poor prognosis of NSCLC, and LGR5⁺ cells in NSCLC were likely to be the cancer cells with stem cell-like properties due to the significant correlation between LGR5 and ALDH1A1.

[Establishment of a sandwich ELISA for testing serum sB7-H3 and detection of sB7-H3 levels in liver disease].

AIM: To establish quantitative ELISA for soluble sB7-H3 and evaluate its clinical application. METHODS: Two mAbs of mouse anti-human sB7-H3(4H7 and 2E6)established by our lab were used. The mAb 4H7 was used as coating antibody and the 2E6 as a sandwich antibody, which recognized a different epitope and was labeled by biotin. The serum sB7-H3 levels of healthy volunteers and patients were measured by this method. RESULTS: Sandwich ELISA was established with the linear range covering from 8.192 ng/L to 2 000 ng/L. The outcomes of repeatability test, recovery test, stability test and specificity test were quite good. The increased levels of sB7-H3 were found in schistosomiasis cirrhosis, the reduced levels in severe hepatitis and the difference in both situations showed significance. CONCLUSION: A specific and sensitive sandwich ELISA is developed, the detection in liver disease indicates that the ELISA kit provides a potential tool in the clinical application.

[Preparation of a 96-microwell plate DNA diagnostic chip for detection of foodborne bacteria and its application in an incident of food poisoning].

OBJECTIVE: To develop a 96-microwell plate DNA diagnostic chip for simultaneous detection of 9 major foodborne bacteria. METHODS: Type-specific PCR primers labeled with biotin and oligonucleotide probes were designed according to the conservative genes of 9 major foodborne bacteria Staphylococcus aureus, Salmonella spp., Escherichia coli O157:H7 (Stx1 and Stx2), Shigella spp., Listeria monocytogenes, Bacillus cereus, Yersinia enterocolitica, Vibrio cholerae and Vibrio parahaemolyticus. A one-tube multiplex PCR system for simultaneous amplification of these bacteria was established, and the DNA probes were spotted and immobilized in the wells of the plate in 5x5 array format. Stable hybridization system between PCR products and oligonucleotide probes in the microwell was established after condition optimization. Alkaline phosphatase-conjugated streptavidin and NBT/BCIP were used to detect the hybridized PCR products. RESULTS: Twenty standard bacteria strains were used to validate the 96 microwell plate DNA diagnostic chip and highly specific and stable experiment results were obtained. Using this chip assay, the causal pathogen Staphylococcus aureus was identified within 12 h after the sampling from an incident of food poisoning, and the result was consistent with that obtained using conventional bacterial culture and biochemical identification. CONCLUSION: The novel 96 microwell plate DNA diagnostic chip allows rapid, accurate, automated and high-throughput bacterial detection and is especially valuable for quick response to such public health emergencies as food poisoning.

[Development a method of quantitative assay for enterovirus 71].

OBJECTIVE: To establish a quantitative assay for enterovirus 71, this can be used in detecting the virus content during vaccine development and production. METHODS: We established the method of quantitative assay for EV71 by using double antibody sandwich ELISA. The sensitivity, accuracy,precision and specificity of the method were evaluated. RESULTS: We developed an ELISA method to quantitative assay for EV71. The quantitation limit of the method is 0.23 microg/ml and the quantitation scope of the method is 7.32-0.23 microg/ml, the coefficient correlation is R2 = 0.9976; The method showed good accuracy, precision and specificity. The recovery is between 90%-110% and the variation coefficient is lower than 10%. CONCLUSION: An ELISA method was developed for the quantitative assay of EV71 virus, which can be used for the rapid quantitative determination of EV71 virus during vaccine development and production.

Study of 1-3-2 type piezoelectric composite transducer array.

Based on a new structure of precise location and decoupling between the transducer elements, high frequency underwater transmission transducer arrays with 4 elements and 8 elements serried uniform linear array were studied, using novel 1-3-2 type piezoelectric composite as the sensing material. There are ceramic framework supports in the transverse and longitudinal directions in 1-3-2 type piezoelectric composite structure; the transducer is free from the influence of outside mechanical impact and the environment temperature change. Transducer and array samples have been designed, fabricated and measured. The resonant frequency is 150 kHz, resonant transmission response greater than 160 dB, and bandwidth is from 140 kHz to 160 kHz. The results indicate that, the transducer array has wide bandwidth, high sensitivity, stabile performance, and good coherence.

A study of a 1-3-2 type piezoelectric composite.

A novel 1-3-2 type piezoelectric composite is developed. There are piezoelectric ceramic framework supports at transverse and longitudinal directions in its structure, so it is free from the influence of any outside mechanic impact and environmental temperature change. The sample characteristic is good coherence. It has many advantages, such as a high piezoelectricity, lower density and acoustic impedance matching with water and tissue easily. Based on Newnham's series and parallel theory of composites, the calculation formulae of dielectric and piezoelectric constants of a 1-3-2 type piezoelectric composite are deduced. The sample's characteristics indicate that a 1-3-2 type piezoelectric composite's theoretic calculation values are in good agreement with the measurement results. Element samples have been designed, manufactured and measured. The results indicate that the resonant frequency, resonant impedance, piezoelectricity and static capacity of these samples are stable. It is easy to design sensor array elements using this material.

[Three-dimensional reconstruction of human organs based on magnetic resonance imaging data].

OBJECTIVE: To build proper three-dimensional models of human organs for radiation dose calculation. METHOD: Human organ models were built by contour reconstruction algorithm conforming to 4 criteria. The contours of the organs were extracted based on the contours of the reconstructed organ surfaces, and the three-dimensional models with smooth surface, exact structure and good visibility were completed by computer graphic techniques such as lighting, smoothing, and normal average. RESULT AND CONCLUSION: Several organ models were successfully reconstructed, suggesting that this method is suitable for building three-dimensional digital human organ models which can meet the need for dose calculation.

Elastic registration of medical images through multiquadric method.

OBJECTIVE: To improve the precision and reliability of elastic registration of the medical images and to simplify the registration process. METHODS: Previous study concerning elastic registration mostly focused on manual selection of the landmarks and then use of adequate interpolating for elastic transformation. The landmarks extraction, however, was prone to error that often showed impact on the registration results, besides the difficulty and time consumption of manual identification of the landmarks. On the basis of Multiquadric method that allowed smooth adjustment of the parameters, we utilized a semi-automatic method to extract the landmarks by combining these 2 steps, and proposed a novel registration method. RESULTS: Using this method for medical image elastic registration, rapid and accurate registration between standard and deformed images was achieved. CONCLUSION: The method proposed presently is accurate, convenient and reliable.

A method for three-dimensional surface reconstruction of volume data field.

OBJECTIVE: To explore a novel method of three-dimensional (3D) reconstruction based on vector field smoothing, for the purpose of 3D surface reconstruction of DICOM format volume data sets. METHODS: 3D external surface of three sets of volume data, namely craniocerebral volume data, pelvis volume data, and rat embryo volume data, were respectively extracted by Marching Cubes algorithm using small triangle flakes to approach the original 3D structure surfaces. Vector field smoothing was performed on the extracted 3D surfaces. The reconstructed 3D structures were rendered from different angles of view through arbitrary rotation. RESULTS: High-quality results of 3D surface reconstruction were obtained for each set of volume data, demonstrating fine 3D surface details and high fidelity. CONCLUSION: This method can improve 3D surface reconstruction from DICOM volume data sets, promising high quality, fidelity and reality.