[The impact of uncertainty resection on the prognosis of non-small cell lung cancer].

Objective: To explore the impact of uncertain resection on postoperative survival in non-small cell lung cancer. Methods: This is a retrospective cohort study. A retrospective analysis was conducted on the data of 477 patients with non-small cell lung cancer who underwent lobectomy in the Department of Thoracic Surgery, the Fourth Hospital of Hebei Medical University from December 2012 to December 2013. There were 302 males and 175 females, aged (59±8) years (range: 27 to 79 years). According to the surgical resection criteria issued by the International Association for the Study of Lung Cancer, the patients were divided into the intact resection group (R0 group, 286 cases) and the uncertain resection group (R (un) group, 191 cases). Clinical data between the two groups were compared using χ2 test, and propensity score matching (PSM) was performed on patients using the R language, with matching variables including gender, age, smoking history, adjuvant therapy, TNM stage, pathological type, and tumor site. The nearest-neighbor method was used for 1∶3 matching and the caliper value was 0.02. The survival curve was plotted using the Kaplan-Meier method and compared using the Log-rank test. The Cox proportional hazards regression model was used to identify risk factors in overall survival (OS). Subgroup analysis was based on TNM staging and mediastinal lymph node metastasis status. Results: In the R (un) group, 68 patients had positive lymph in the highest group and 129 patients did not undergo complete dissection of the mediastinal lymph nodes. The baseline data for the R0 group and the R (un) group were corrected using PSM, and a total of 369 patients were successfully matched, including 227 cases in the R0 group and 142 cases in the R (un) group. After PSM, the 5-year survival rates of the R0 group and the R (un) group were 64.3% and 52.1%, respectively (P=0.021). The 5-year survival rates of stage Ⅰ, Ⅱ, and Ⅲ patients were 85.2%, 65.9%, and 34.8%, respectively (P<0.01). TNM stage (χ2=46.913, P<0.01), pathological classification of adenosquamous cell carcinoma (HR=5.970, 95% CI: 3.117 to 11.431, P<0.01) and R (un) resection (HR=1.512, 95% CI: 1.065 to 2.147, P=0.021) were prognostic factors for postoperative survival. Subgroup analysis showed that in stage Ⅲ patients, 5-year survival rates of the R0 group and the R (un) group after resection were 45.8% and 9.5%, respectively (P=0.002). Among patients with mediastinal lymph node metastasis, 5-year survival rates of the R0 group and the R (un) group were 50.6% and 7.1%, respectively (P<0.01). Conclusions: TNM staging, pathological type, and R (un) resection are prognostic factors for overall postoperative survival in non-small cell lung cancer. In stage Ⅰ and Ⅱ patients, R (un) is not a prognostic factor for postoperative survival of non-small cell lung cancer. In patients with stage Ⅲ and mediastinal lymph node metastasis, R (un) is a prognostic factor for non-small cell lung cancer after surgery.

The Relationship between Choroidal Thickness and Liver Damage in Simple Auto-immune Hepatitis Patients.

BACKGROUND: There was no sufficient clinical evidence on the relationship between auto-immune hepatitis (AIH) and risk of eye illness, except 11 uveitis cases where related AIH is reported currently. AIM: To determine the relationship between choroidal thickness (ChT) and liver damage in simple AIH patients without ocular symptoms after oral prednisone treatment. PATIENTS AND METHODS: This prospective observational study included simple AIH patients. The patients' ChT was measured by swept-source (SS)-optical coherence tomography (OCT), and the liver damage was evaluated by alanine aminotransferase (ALT) and aspartate aminotransferase (AST). ChT and liver functions were assessed prior to and after treatment. Then comparisons were made prior to and post treatment. The relationships between biochemical indexes of liver injury and ChT were evaluated after a mean (SD) of 24 (1.28) weeks of regular oral prednisone. RESULTS: A total of 35 patients (31 females, aged 45.66 ± 11.62 years) were included. After treatment, ChT was significantly increased in all sectors (including the center sector, superior inner sector, inner nasal sector, inferior inner sector, inner temporal sector, superior outer sector, outer nasal sector, inferior outer sector, and outer temporal sector) (all P < 0.001). After treatment, both ALT (51.34 ± 44.16 vs 255.06 ± 107.84, P < 0.001) and AST (38.66 ± 27.12 vs 164.89 ± 85.58, P < 0.001) were significantly decreased. The increase of ChT in all sectors was significantly related to the decrease of ALT and AST (all P < 0.001). CONCLUSION: The improvement of ChT might reflect the remission of liver damage in simple AIH patients without ocular symptoms during oral prednisone treatment.

[Pancreas multidisciplinary team optimizes the diagnosis and treatment of pancreas-related diseases and improves the prognosis of pancreatic cancer patients].

Objectives: To evaluate the role of pancreas multidisciplinary team(MDT) clinic in the diagnosis of pancreatic diseases,patient compliance with MDT advice,and the impact of MDT on the postoperative survival of patients with pancreatic cancer. Methods: The study included 927 patients(554 males,373 females,aged (58.1±13.3)years (range: 15 to 89 years)) that had visited the pancreas MDT clinic of Zhongshan Hospital from May 2015 to December 2021,and 677 patients(396 males, 281 females, aged (63.6±8.9)years(range: 32 to 95 years)) who underwent radical surgery and with pathologically confirmed pancreatic adenocarcinoma from January 2012 to December 2020,of whom 79 patients had attended the pancreas MDT. The clinical and pathological data were collected and analyzed retrospectively. Diseases were classified in accordance with 2010 WHO classification of tumors of the digestive system and usual clinical practices. The Kaplan-Meier method was used for drawing the survival curve and calculating the survival rate. The univariate analysis was done by Log-rank test and the multivariate analysis was done by COX proportional hazards model. Survival rates were compared using χ2 test. Results: Among the 927 patients that had visited the MDT clinic,233 patients(25.1%) were referred due to undetermined diagnosis. A direct diagnosis was made in 109 cases (46.8%,109/233) by the MDT clinic, of which 98 were consistent with the final diagnosis,resulting in an accuracy of 89.9%(98/109). The direct diagnosis rate in the recent years(36.6%(41/112),from June 2019 to December 2021) decreased compared to that in the previous years(56.2%(68/121),from May 2015 to May 2019),yet the accuracy in the recent years(90.2%,37/41) was basically the same as before (89.7%,61/68). The rate of compliance of the entire cohort was 71.5%(663/927), with the compliance rate in the recent two and a half years(81.4%,338/415) remarkably higher than that in the previous four years(63.4%,325/512). Patients with pancreatic cancer that attended the MDT exhibited a trend toward longer median postoperative survival than patients that did not attend the MDT,but the difference was not statistically significant(35.2 months vs.30.2 months,P>0.05). The 1-year and 3-year survival rates of patients that attended the MDT were significanly higher than patients that did not attend the MDT(88.6% vs. 78.4%,P<0.05;32.9% vs. 21.9%,P<0.05,respectively),but the 5-year survival rate was not statistically different(7.6% vs. 4.8%,P>0.05). Conclusions: The pancreas MDT clinic is an accurate and convenient way to diagnose intractable pancreatic diseases,and in the recent years the patients' compliance rate with MDT advice has increased. Pancreatic cancer patients that have attended the MDT have higher 1-year and 3-year postoperative survival rates,but the long-term survival benefits of MDT still needs to be proved by clinical studies on a larger scale.

Molecular Characterisation, Tissue Distribution, and Expression Profiling of the Cathepsin B Gene during Ovarian Follicle Development in Geese.

1. Although there is evidence that Cathepsin B (CTSB) regulates the degradation and absorption of yolk precursors during avian ovarian follicle development, nothing is known about its molecular characteristics, tissue distribution or expression profiles in goose ovarian follicular compartments.2. The intact 1023 bp coding sequence of the goose CTSB gene was obtained for the first time. It encoded a polypeptide of 340 amino acids (AA) containing two conserved functional domains (i.e., Propeptide_C1 and Peptidase_C1A_Cathpsin B) and three active amino acid residues (+108, +279, and +299). Both the nucleotide and AA sequences of goose CTSB gene showed more than 90% similarity with its respective homologs from other avian species.3. The qRT-PCR results showed that CTSB mRNA was ubiquitously expressed in all examined goose tissues, with moderate to high levels in the reproductive organs including the ovarian stroma and oviduct.4. Expression of goose CTSB mRNA in the granulosa layers increased gradually from the 2-4 mm F5 follicles but declined to relatively low levels in the F4-F1 follicles while remaining statistically unchanged in the theca layers throughout follicle development.5. High sequence similarity of goose CTSB gene to other avian species suggested functional conservation of avian CTSB genes, and its fluctuating levels in the granulosa layers may be associated with the orderly progression of goose follicle development. These data laid a foundation for further elucidating the role of CTSB in the avian ovary.

MiR-181a-5p inhibits goose granulosa cell viability by targeting SIRT1.

1. Granulosa cells (GCs) are involved in folliculogenesis, follicular development, and atresia. Previous studies have shown that microRNA-181a-5p (miR-181a-5p) and sirtuin 1 (SIRT1) are involved in GC proliferation and apoptosis, and SIRT1 has been predicted as one target of miR-181a-5p. However, there are few studies with poultry.2. Quantitative real-time PCR (qRT-PCR) was used to detect the expression level of miR-181a-5p in granulosa layers during geese ovarian follicular development. A methyl thiazolyl tetrazolium (MTT) assay was performed to assess the viability of geese granulosa cells treated with miR-181a-5p mimic or inhibitor. The binding sites between the SIRT1 3'-UTR region and miR-181a-5p were evaluated using a luciferase reporter assay system. SIRT1 mRNA levels were detected using qRT-PCR after transfection with miR-181a-5p mimic and inhibitor.3. The miR-181a-5p suppressed geese GC viability and regulated the mRNA expression of viability-related genes in geese GCs. SIRT1 was a target gene of miR-181a-5p and miR-181a-5p suppressed its mRNA expression.4. The miR-181a-5p may target and inhibit SIRT1 expression, thus suppressing GC viability by regulating viability-related key genes.

[Changes of serum markers in acute liver injury induced by oral 1, 2-dichloropropane in BALB/cA-nu mice].

Objective: To investigate the characteristics of inflammatory response in BALB/cA-nu mice induced by oral 1, 2-dichloropropane (1, 2-DCP) , and to provides theoretical reference for further study of subchronic, chronic toxicity and carcinogenic mechanism. Methods: From October 2018, Clean grade healthy BALB/cA-nu mice were randomly divided into 5 groups with 10 mice in each group. And 860, 1150, 1500, 1950, 2535 mg/kg 1, 2-DCP were given by gavage respectively. Meanwhile, blank group and solvent control group (corn oil) were set up. Blood samples were collected from eyeballs and liver and bile tissues were collected for histopathological examination within 24 hours after exposure. The expression of alanine aminotransferase (ALT) , total bilirubin (TBLI) , C-reactive protein (CRP) , interleukin-6 (IL-6) , tumor necrosis factor-α (TNF-α) and tumor necrosis factor-ß (TNF-ß) were detected by enzyme-linked immunosorbent assay. Results: With the increase of the dose of 1, 2-DCP, the number of microbubbles in liver cells and the infiltration of inflammatory cells increased gradually. No pathological changes were found in the gallbladder. Compared with the blank group and solvent control group, the content of serum ALT in each exposure group was increased, the serum levels of IL-6 and TNF-ß in 860, 1150, 1950 and 2350 mg/kg exposure groups were increased, the serum TNF-α and TBLI levels in 1 950, 2535 mg/kg groups were significantly higher (P<0.05) . The levels of ALT, TBLI and TNF-ß in serum of female mice were significantly different (P<0.05) . There were significant differences in ALT, TBLI, CRP, IL-6, TNF-α, TNF-ß in serum of male mice (P<0.05) . Conclusion: Oral 1, 2-DCP may cause acute liver injury in BALB/cA-nu mice and increase the expression of serum inflammatory factors. Moreover, the types of inflammatory factors activated in male mice are more than those in female mice.

Long noncoding RNA SNHG12 promotes the progression of cervical cancer via modulating miR-125b/STAT3 axis.

Increasing evidence showed that long noncoding RNAs (lncRNAs) played an important role in the occurrence and development of tumors. To date, lncRNA small nucleolar RNA host gene 12 (SNHG12) has revealed an oncogenic role in various tumors. However, the role of SNHG12 in cervical cancer is still unclear. Therefore, we focused on the biological function and molecular mechanism of SNHG12 in the tumorigenesis of cervical cancer. In this study, the expression of miR-125b was observably downregulated in cervical cancer cells. Meanwhile, the expression of SNHG12 was obviously upregulated in cervical cancer cell lines (HeLa, SiHa, Caski, C4-1, and C33A) compared with the immortalized cervical epithelial cells. The further assay showed that miR-125b was a target of SNHG12 in cervical cancer. Moreover, a negative relationship between miR-125b and SNHG12 was found in cervical cancer. In addition, SNHG12 inhibition restrained the proliferation, migration, and invasion of cervical cancer cells. Meanwhile, miR-125b mimics repressed the expression of signal transducer and activator of transcription 3 (STAT3). The further assay showed that STAT3 was a target of miR-125b in cervical cancer. In addition, sh-STAT3 repressed the migration and invasion of cervical cancer cells. Furthermore, it showed that miR-125b inhibitors reversed STAT3 expression restrained by the reduction of SNHG12 expression. In general, SNHG12 modulated STAT3 by sponging miR-125b in cervical cancer and played an important role in the development of cervical cancer.

[The value of preoperative CA19-9 combined with platelet-to-lymphocyte ratio in predicting invasive malignancy in intraductal papillary mucinous neoplasms].

Objective: To explore preoperative predictive markers for invasive malignancy in intraductal papillary mucinous neoplasm(IPMN). Methods: The retrospective case-controlled study was adopted.Seventy-nine patients who underwent surgery and with pathologically confirmed IPMN from January 2005 to December 2014 at Department of Pancreatic Surgery, Zhongshan Hospital Fudan University were enrolled.Forty-six patients were male and 33 were female,with an average age of (62.9±8.9)years (range:37-82 years).Tumor sites:56 tumors were located at the head of the pancreas,22 were located at the body and tail of the pancreas,and 1 was located across the whole pancreas.Surgical procedures: 51 patients underwent pancreaticoduodenectomy, 22 patients underwent distal pancreatectomy, 4 patients underwent segmental pancreatectomy and 2 patients underwent total pancreatectomy.IPMNs were classified into non-invasive lesions and invasive carcinomas according to the histopathological findings of the tumor.Thirty-two tumors were non-invasive lesions and 47 were invasive carcinomas.The preoperative findings were compared between patients with non-invasive IPMN and patients with invasive carcinoma by univariate analysis using t test and χ(2) test accordingly,and factors with statistically significance were subsequently submitted to multivariate analysis. Results: Univariate analysis showed that tumor size(P=0.022), carcinoembryonic antigen(P=0.012), CA19-9(P=0.011), lymphocytes(P=0.034), neutrophil-to-lymphocyte ratio(P=0.010)and platelet-to-lymphocyte ratio(PLR)(P=0.004)were predictive markers with statistical significance.Multivariate analysis showed that CA19-9(P=0.012)and PLR(P=0.025) were independent predictive markers for invasive malignancy in IPMN.The area under curve of the combination factor of CA19-9 and PLR(0.864) was larger than that of CA19-9(0.806) or PLR(0.685) alone, and all the authentic indicators of the combination factor were better than those of each alone. Conclusions: CA19-9 and PLR are independent predictive markers for invasive malignancy in IPMN.The combination of CA19-9 and PLR has improved efficacy than each alone.

[Evaluating the level of occupational stress and its influence factors among traffic police in a district in Shanghai].

Objective: To investigate the high occupational stress and its influencing factors in traffic police in Shanghai. Methods: 728 traffic police were selected as the study subjects, and the《Occupational Health Questionnaire》was used to investigate and evaluate their job demand-control (JDC) and effort-reward imbalance model (ERI) occupational stress situation respectively. The related influencing factors were analyzed. Results: The prevalence rates of high occupational stress in JDC and ERI models were 74.6% (543/728) and 51.5% (375/728) . The influencing factors of JDC were education, marriage, average weekly hours (χ(2)=16.82, 10.04, 18.71, P<0.05) , and The influencing factors of ERI were gender, age, marriage, real monthly income level, education, work experience, and average weekly hours (χ(2)=7.02, 26.18, 6.73, 50.42, 4.75, 26.61, 112.98, 6.19, P<0.05) . The JDC multivariate logistic analysis indicated that the risk of occupational stress of married police was 2.81 times as high as that of Unmarried ones. The risk of occupational stress of traffic police with more education was 1.92 times as high as that of low eduacation, average weekly working 41-50 hours and≥51 hours was 2.53, 3.12 times as high as that of ones with average working 40 hours, respectivly. Meanwhile, the ERI multivariate logistic analysis indicated that high income level is the protective factor of occupational stress. The traffic police with 15-<20 working years were more likely to occur higher occupational stress. The traffic police with the more average weekly hours had greater possibility of higher occupational stress. Conclusion: The main influencing factors of JDC and ERI are marriage, real monthly income level, education, work experience, and average weekly hours.

Gene expression profiles indicate tissue-specific obesity regulation changes and strong obesity relevant tissues.

BACKGROUND: With the growing evidence that other tissues, apart from adipose, could have strong relevance to obesity, it is necessary to comprehensively understand the relationship between obesity and other tissues, and to point out the most relevant tissues. METHODS: There were 549 participants with 20 different tissue types involved in this study. We firstly employed both Spearman's correlation test and WGCNA (weighted correlation network analysis) to identify body mass index (BMI)-related genes. Subsequently, we performed enrichment analyses with obesity genes and pathways to see the different regulation patterns among tissues. In addition, we compared obesity genes identified by genome-wide association studies (GWAS) with BMI-related genes to find the overlapping proportion in each tissue. Finally, we integrated preceding results to identify six strong obesity relevant tissues and indicate three categories to represent different obesity relevant tissues. RESULTS: Statistical analyses revealed diverse BMI-related genes and tissue-specific enrichment patterns among tissues. Comparison between BMI-related genes and GWAS findings showed tissue-specific expression changes of GWAS genes. Ultimately, six tissues that showed predominant performance in enrichment analyses and significantly embraced GWAS genes were referred to as strong obesity relevant tissues, including adipose, esophagus, nerve, pancreas, pituitary and skin. We also proposed three categories to represent different obesity relevant tissues. CONCLUSIONS: We performed the first study to investigate the BMI-related gene expression changes across 20 tissues at the same time. With valid data analyses and comparison with GWAS findings, our study provides a holistic view of how different tissues correlate with obesity, and proposes target tissues for obesity pathogenesis investigation.

Post hoc support vector machine learning for impedimetric biosensors based on weak protein-ligand interactions.

Impedimetric biosensors for measuring small molecules based on weak/transient interactions between bioreceptors and target analytes are a challenge for detection electronics, particularly in field studies or in the analysis of complex matrices. Protein-ligand binding sensors have enormous potential for biosensing, but achieving accuracy in complex solutions is a major challenge. There is a need for simple post hoc analytical tools that are not computationally expensive, yet provide near real time feedback on data derived from impedance spectra. Here, we show the use of a simple, open source support vector machine learning algorithm for analyzing impedimetric data in lieu of using equivalent circuit analysis. We demonstrate two different protein-based biosensors to show that the tool can be used for various applications. We conclude with a mobile phone-based demonstration focused on the measurement of acetone, an important biomarker related to the onset of diabetic ketoacidosis. In all conditions tested, the open source classifier was capable of performing as well as, or better, than the equivalent circuit analysis for characterizing weak/transient interactions between a model ligand (acetone) and a small chemosensory protein derived from the tsetse fly. In addition, the tool has a low computational requirement, facilitating use for mobile acquisition systems such as mobile phones. The protocol is deployed through Jupyter notebook (an open source computing environment available for mobile phone, tablet or computer use) and the code was written in Python. For each of the applications, we provide step-by-step instructions in English, Spanish, Mandarin and Portuguese to facilitate widespread use. All codes were based on scikit-learn, an open source software machine learning library in the Python language, and were processed in Jupyter notebook, an open-source web application for Python. The tool can easily be integrated with the mobile biosensor equipment for rapid detection, facilitating use by a broad range of impedimetric biosensor users. This post hoc analysis tool can serve as a launchpad for the convergence of nanobiosensors in planetary health monitoring applications based on mobile phone hardware.

[Effect of Gas6 in silica-induced inflammation on differentiated human acute monocytic leukemia (THP-1) macrophages].

Objective: To investigate the modulation role of Gas6 in silica-induced inflammatory effect on human macrophages. Methods: Differentiated THP-1 macrophages were exposed to different concentrations of silica for 6 h and 24 h. Additionally, silica-activated macrophages were treated with different concentrations of recombine human Gas6 and Gas6 antibody respectively. Cell viabilities were determined by CCK-8 kit. Expression levels of Gas6 and inflammatory cytokines (TNF-α, IL-1ß and IL-6) were measured by ELISA assay kits. Results: Silica particles induced clear dose-dependent decreases of cell viability and Gas6 expression at both 6 h and 24 h. The cell viability of 24 h is lower than 6 h at the same concentration of silica (P<0.05). Furthermore, silica activated macrophages treated with Gas6 antibody induced significant decreases of Gas6 both at 6 h and 24 h (P<0.05). After pretreated with various concentrations of Gas6 antibody, silica induced higher expressions of inflammatory cytokines (TNF-α, IL-1ß, IL-6) in dose-dependent manners at two time points. Addition of exoge-nous Gas6 significantly suppressed silica-induced inflammatory cytokines concentrations mentioned above in the cell culture supernatants in clear dose-dependent manners. Conclusion: Exogenous Gas6 could inhibit the secre-tion of inflammatory cytokines in macrophages, while the block of Gas6 might enhance this inflammation.

pulSED: pulsed sonoelectrodeposition of fractal nanoplatinum for enhancing amperometric biosensor performance.

For the first time, we combine pulsed electrodeposition with out-of-phase pulsed sonication for controlled synthesis of fractal nanoplatinum structures as the transducer layer in electrochemical sensing. We develop and test this technique, called bimodal pulsed sonoelectrodeposition (pulSED), as a simple approach for creating highly conductive transducer nanometals for use in sensing and biosensing. We first compared the efficiency of pulSED nanoplatinum to other pulsed electrodeposition techniques, and then explored the effect of duty cycle and plating time on electroactive surface area and nanoparticle size/morphology. The developed pulSED nanoplatinum displayed fractal features with a relatively homogenous size distribution (26.31 ± 1.3 nm) and extremely high electroactive surface (0.28 ± 0.04 cm(2)) relative to other electroplating techniques (up to one order of magnitude higher). A high duty cycle (900 mHz) promotes formation of stable nanostructures (including fractal nanostructures) and reduces amorphous structure formation due to bubble cavitation and enhanced mass transport of metal ions to the electrode surface. To demonstrate the applicability of the pulSED technique, non-enzymatic and enzymatic sensors were developed for measuring hydrogen peroxide and glucose. The sensitivity for non-enzymatic peroxide sensing (3335 ± 305 µA cm(-2) mM(-1)), non-enzymatic glucose sensing (73 ± 14 µA cm(-2) mM(-1)) and enzymatic glucose biosensing (155 ± 25 µA cm(-2) mM(-1)) was higher than, or similar to, other nanomaterial-mediated amperometric sensors reported in the literature. The pulSED technique is a one pot method for tunable synthesis of nanometal structures as a transducer layer in electrochemical sensing and biosensing that requires no precursors or capping agents, and can be carried out at room temperature with inexpensive hardware.

A chemical-structural model for coherent martensite/parent interface in Mn-based antiferromagnetic shape memory alloys.

The martensite/parent coherent interface of Mn-based shape memory alloys (SMAs) is a significant part in the research of their martensitic transformation, reversible shape memory effect and magnetic shape memory effect. In the present work, a chemical-structural model was proposed to calculate the martensite/parent coherent interfacial energy of Mn-X (X = Cu, Fe) alloys. In this model, the coherent heterophase interfacial energy consists of chemical and structural parts. Resulting from the formation process of the heterophase interface, the chemical interfacial energy is expressed as the incremental value of bond energy, while the structural part is obtained by calculating the interfacial strain energy. The results show that the structural interfacial energy plays the chief role in the total interfacial energy, and the total interfacial energy decreases as the temperature rises when the alloy composition is fixed. In addition, the preferred orientation has noteworthy influence on the total interfacial energy. Using the proposed model, interfacial energy, interfacial entropy, interfacial enthalpy and interfacial heat capacity are found to be correlated with temperature and interface preferred orientation. Furthermore, the influences of alloy composition, modulus softening, and the index of the habit plane on the results were discussed.

Effects of grafting on key photosynthetic enzymes and gene expression in the citrus cultivar Huangguogan.

Grafting influences scion photosynthetic capacity and fruit quality. Ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco), which strongly affects photosynthetic rate, and Rubisco activase (RCA), which regulates Rubisco activity, are two key photosynthetic enzymes. However, little information is available regarding the effect of grafting on the concentration and expression of Rubisco and RCA in the citrus cultivar Huangguogan. The objective of this study was to investigate the effect of grafting Huangguogan plants onto trifoliate orange, tangerine, and orange on: 1) the concentration of Rubisco and RCA; 2) the mRNA levels of rbcL, rbcS, and rca; and 3) fruit quality. Overall, the results showed that when Huangguogan plants budded on tangerine and orange, they had better fruit quality, while on trifoliate orange they had higher Rubisco concentration. Tangerine and orange are probably the most suitable rootstocks for Huangguogan plants given the environmental conditions of Sichuan Province, China.

[Treatment of liver cancer in vitro and in mice by monoclonal antibody targeting epithelial specific antigen-positive liver cancer stem cells in combination with cisplatin].

OBJECTIVE: To investigate the biological characteristics of monoclonal antibodies against human liver cancer stem cells and its therapeutic effect in combination with cisplatin in the treatment of hepatocellular carcinoma. METHODS: Cell culture in serum-free medium and PKH26 staining were used to determine the existence of cancer stem cells in human liver Bel7402-V3 cell line. The co-expression of antigen recognized by monoclonal antibody (McAb) 15D2 and epithelial specific antigen (ESA) and PKH26-positive cells in the Bel7402-V3 cells were detected by immunofluorescence assay. Serum-free suspension culture was used to detect the self-renewal ability of 15D2-positive Bel7402-V3 cells sorted by flow cytometry and the effect of 15D2 on the self-renewal ability of Bel7402-V3 cells. The effect of 15D2 on cisplatin resistance in the cells was examined by CCK8 method. The inhibitory effect of 15D2 combined with cisplatin on the transplanted tumor growth in mice was also observed. RESULTS: Single PKH26-positive cells were observed in the Bel7402-V3 cell spheroids cultured for 11 days. Immunofluorescence assay showed that the 15D2-recognized antigen could be conjugated with PKH26 and ESA and co-localized on Bel7402-V3 cells. The spheroid formation rate of 15D2-positive cells in serum-free medium was significantly higher than that of 15D2-negative cells [(30.4±3.4)% vs. (8.8±1.8)%, P<0.01]. The cisplatin resistance of 15D2-positive cells was obviously higher than that of 15D2-negative cells (IC50: 1.014 µmol/L vs. 0.365 µmol/L). McAb 15D2 significantly suppressed the spheroid formation of Bel7402-V3 cells, with an inhibition rate of 37.5%. McAb 15D2 also notably inhibited the cisplatin resistance of Bel7302-V3 cells. The IC50 was 0.211 µg/ml in the 15D2 group and 0.325 µg/ml in the control group. The mouse experiment showed that the tumor growth rates of 50 mg/kg, 25 mg/kg and 12.5 mg/kg 15D2-treatment groups were 82.6%, 71.4% and 60.0%, respectively; that of the 50 mg/kg 15D2 + cisplatin group was 91.0%, and that of the cisplatin monotherapy was 56.7%. CONCLUSION: McAb 15D2 is a functional monoclonal antibody targeting liver cancer stem cells, which could be a potential monoclonal antibody drug for the stem cell-targeted therapy of liver cancer.

A comparative study of graphene-hydrogel hybrid bionanocomposites for biosensing.

Hydrogels have become increasingly popular as immobilization materials for cells, enzymes and proteins for biosensing applications. Enzymatic biosensors that utilize hydrogel as an encapsulant have shown improvements over other immobilization techniques such as cross linking and covalent bonding. However, to date there are no studies which directly compare multiple hydrogel-graphene nanocomposites using the same enzyme and test conditions. This study compares the performance of four different hydrogels used as protein encapsulants in a mediator-free biosensor based on graphene-nanometal-enzyme composites. Alcohol oxidase (AOx) was encapsulated in chitosan poly-N-isopropylacrylamide (PNIPAAM), silk fibroin or cellulose nanocrystals (CNC) hydrogels, and then spin coated onto a nanoplatinum-graphene modified electrode. The transduction mechanism for the biosensor was based on AOx-catalyzed oxidation of methanol to produce hydrogen peroxide. To isolate the effect(s) of stimulus response on biosensor behavior, all experiments were conducted at 25 °C and pH 7.10. Electroactive surface area (ESA), electrochemical impedance spectroscopy (EIS), sensitivity to methanol, response time, limit of detection, and shelf life were measured for each bionanocomposite. Chitosan and PNIPAAM had the highest sensitivity (0.46 ± 0.2 and 0.3 ± 0.1 µA mM(-1), respectively) and electroactive surface area (0.2 ± 0.06 and 0.2 ± 0.02 cm(2), respectively), as well as the fastest response time (4.3 ± 0.8 and 4.8 ± 1.1 s, respectively). Silk and CNC demonstrated lower sensitivity (0.09 ± 0.02 and 0.15 ± 0.03 µA mM(-1), respectively), lower electroactive surface area (0.12 ± 0.02 and 0.09 ± 0.03 cm(2), respectively), and longer response time (8.9 ± 2.1 and 6.3 ± 0.8 s, respectively). The high porosity of chitosan, PNIPAAM, and silk gels led to excellent transport, which was significantly better than CNC bionanocomposites. Electrochemical performance of CNC bionanocomposites were relatively poor, which may be linked to poor gel stability. The differences between the Chitosan/PNIPAAM group and the Silk/CNC group were statistically significant (p < 0.05) based on ANOVA. Each of these composites was within the range of other published devices in the literature, while some attributes were significantly improved (namely response time and shelf life). The main advantages of these hydrogel composites over other devices is that only one enzyme is required, all materials are non-toxic, the sensor does not require mediators/cofactors, and the shelf life and response time are significantly improved over other devices.

Resveratrol appears to protect against oxidative stress and steroidogenesis collapse in mice fed high-calorie and high-cholesterol diet.

The detrimental effects on Leydig cells steroidogenesis in mice on high-calorie and high-cholesterol diet (HCD) were determined, and the possible protection conferred by resveratrol supplementation was investigated. Male C57BL/6J mice were fed high-calorie and alone (HCD group) or with resveratrol supplementation (HCD + Res group) for 18 weeks. Male C57BL/6J mice fed standard diet without or with the same dose of resveratrol served as controls. At the end of the experiment, there were significant declines of serum testosterone and luteinising hormone (LH) in HCD group as compared to controls. In line with the hormone alterations, the expressions of StAR and steroidogenic enzymes in testicular tissues were significantly down-regulated in HCD group. Resveratrol supplementation could significantly improve expressions of StAR and steroidogenic enzymes, and increase serum testosterone and LH concentrations in HCD + Res group. Mice in HCD group also showed a statistically significant down-regulation in the mRNA expressions of MnSOD and GPx4. Resveratrol supplementation improved testicular MnSOD and GPx4 expression in comparison with HCD group. We propose that resveratrol may attenuate detrimental effects on Leydig cells steroidogenesis in HCD-fed mice, and its upregulations of antioxidant defence mechanisms and LH level may play a role in its protection. Our data suggest resveratrol appears to have the potential for therapeutic approaches targeting male obesity-associated secondary hypogonadism.

[Clinical application of lateral great toe flap in decoratively reconstruction of thumb pulp defect].

OBJECTIVE: To investigate the validity and the outcome of a new surgery method that utilizes lateral great toe flap to decoratively reconstruct the thumb pulp loss. METHODS: From Jan. 2009 to Jan. 2014, 22 cases with unilateral thumb pulp defect were included, for whom, lateral great toe flap was deployed to reconstruct the thumb. Blood circulation was re-established by the anastomosis of the digital vessels of toes and fingers in 18 cases, the other 4 cases were conducted by a dorsal metatarsal artery and vein anastomosed to the deep branch of the radial artery and the dorsal metacarpal vein respectively; as for the nerve repair, toe-to-finger digital nerve anastomosis was performed. The donor sites were covered by full-thickness inguinal skin grafts. RESULTS: All the 22 flaps survived without complications. With 6 to 12 months follow-up, the surviving fingers all presented aesthetically pleasant appearance with vivid loops and whorls. Normal functions like sweating were restored as well, and the two-point-discrimination ranged 3.0 to 6.0 mm (4.2 mm in average). The skin grafts to the donor sites healed uneventfully, the appearance and functions of the feet were unaffected. CONCLUSION: Lateral toe flap should be the first-line choice for reconstructing thumb defect, as it conveniently restores both functions and outlook.

Tim-3 expression on peripheral monocytes and CD3+CD16/CD56+natural killer-like T cells in patients with chronic hepatitis B.

Hepatitis B virus (HBV) infection is one of the major causes of chronic liver inflammation. Tim-3 acts as a negative regulatory molecule and plays a critical role in immune tolerance. In the current study, we investigated Tim-3 expression on peripheral monocytes and CD3+CD16/CD56+ natural killer like T (NKT-like) cells in chronic hepatitis B (CHB) patients. Peripheral blood mononuclear cells (PBMCs) were isolated from 52 CHB patients and 60 healthy controls. Tim-3+CD14+ cells and Tim-3+CD3+CD16/CD56+ cells were analyzed by flow cytometry. Results showed that expression of Tim-3 was significantly increased on both the monocytes and NKT-like cells in CHB patients than in controls (P = 0.002 and P < 0.001, respectively). Tim-3 levels on monocytes and NKT-like cells were further upregulated in patients with acute-on-chronic liver failure (ACLF). In addition, we assessed the correlation of Tim-3 expression with levels of alanine aminotransferase (ALT) and tumor necrosis factor alpha (TNF-α). Data revealed that Tim-3 expression on both monocytes and NKT-like cells was positively correlated with level of ALT (r = 0.59, P < 0.001, and r = 0.60, P < 0.001, respectively), whereas Tim-3 expression on NKT-like cells was negatively correlated with serum level of TNF-α (r = -0.54, P < 0.001) in CHB patients. Our results suggest that Tim-3 may play important roles in the pathogenesis of CHB.