Reveal the relationship between the quality and the cuticle composition of Satsuma mandarin (Citrus unshiu) by postharvest heat treatment.

To investigate the influence of heat treatment (HT) on Satsuma mandarin fruit's postharvest quality and cuticle composition, we immersed the fruit for 3 min in hot water at 52°C and subsequently stored them at room temperature (25°C) for 28 days, and fruit quality parameters, such as good fruit rate, weight loss rate, firmness, total soluble solids, total titratable acidity, and ascorbic acid content, were monitored. Additionally, changes in the peel's cuticle composition were analyzed, and wax crystal morphologies on the fruit surface were examined using scanning electron microscopy (SEM). The findings revealed that appropriate HT effectively preserved fruit quality. The main compositions of wax and cutin on the fruit's surface remained consistent between the HT and the CK during storage. The total content of wax and cutin initially increased, peaking on the 14th day of storage, and then decreased, falling below the levels observed on day 0. Notably, the total amount of cutin in the HT group exceeded that of the control group. Specifically, ω-hydroxy fatty acids with mid-chain oxo groups and mid-oh-ω-hydroxy fatty acids constituted approximately 90% of the total cutin content. Moreover, the HT group exhibited higher (p < 0.05) total wax content in relation to the control. Fatty acids and alkanes were the predominant components, accounting for approximately 87.5% of the total wax. SEM analysis demonstrated that HT caused wax crystals to melt and redistribute, effectively filling wax gaps. It suggests that HT holds promising potential as a green, safe, and eco-friendly commercial treatment for preserving the postharvest quality of Satsuma mandarin. PRACTICAL APPLICATION: In this study, Satsuma citrus (Citrus unshiu) underwent heat treatment (HT) and was subsequently preserved at room temperature (25°C) for 28 days. The findings revealed that HT significantly improved fruit quality compared to the control group. These findings provide valuable insights into the advancement of eco-friendly and pollution-free citrus preservation methods, offering essential strategies and process parameters for their practical application.

Controlled Atmosphere Improves the Quality, Antioxidant Activity and Phenolic Content of Yellow Peach during the Shelf Life.

Controlled atmosphere (CA) has been demonstrated to maintain the shelf-life quality of fruits, but its effect on the antioxidant activities and phenolic content of yellow peach is not comprehensive. This study analyzed the role of CA on the quality of shelf period, phenolic content and antioxidant activity of "Jinxiu" yellow peach. Yellow peach was left under specific aeration conditions (3.5-4% CO2, 2-3% O2, 92-95.5% N2, 1 ± 0.5 °C) and the control (1 ± 0.5 °C) for 21 d, to observe changes in physiological parameters of the fruit during 10 d of the shelf life (25 ± 1 °C). The result showed that CA reduced the weight loss rate (WLR), decay rate (DR), and browning index (BI) of yellow peaches. Furthermore, the CA held a high level of total flavonoid content (TFC), total phenol content (TPC) and phenolic content in the fruit. Antioxidant analysis showed that polyphenol oxidase (PPO) enzyme activity was lower and free radical scavenging capacity (DPPH, ABTS, and FRAP) and antioxidant enzyme activities (POD and PAL) were higher in the CA group. Combining the results of significance analysis, correlation analysis, principal component analysis (PCA) and hierarchical cluster analysis (HCA) clearly identified the differences between the CA group and the control group. The results showed that the CA could maintain higher phenolic content and reduce the oxidation of yellow peach fruit and enhance fruit quality by affecting the antioxidant activities of yellow peach.

Preparation, characterization, and antifungal property of the inclusion complex of Litsea cubeba essential oil/hydroxypropyl-ß-cyclodextrin and its application in preservation of Shatang mandarin.

To explore the potential application of plant essential oil in the postharvest preservation of fruits, the inclusion complex (IC) of Litsea cubeba essential oil (LCEO) with hydroxypropyl-ß-cyclodextrin (HPßCD), prepared by the saturated aqueous solution method, was studied. LCEO/HPßCD-IC was characterized by fourier transform infrared spectroscopy (FT-IR), X-ray diffraction (XRD), scanning electron microscopy (SEM), particle size distribution, and thermogravimetric-differential scanning calorimetry (TG-DSC) analysis. The formation of LCEO/HPßCD-IC was confirmed, and the volume average particle diameter was 24.376 µm. Due to the inclusion of HPßCD, the volatility of LCEO was significantly reduced and the thermal stability was significantly improved. In addition, the antifungal activities of the LCEO ICs were compared, and LCEO/HPßCD-IC was more effective against the citrus postharvest pathogens (P. italicum and G. citri-aurantii). The effects of the LCEO ICs on the postharvest quality of Shatang mandarin were studied. Compared with the control group (CK) and LCEO/ßCD-IC group, the LCEO/HPßCD-IC group showed a significant delay in the decrease of good fruit rate, hardness, total soluble solids (TSSs), and Vitamin C (Vc) content, with a lower weight loss rate of Shatang mandarin. Therefore, LCEO/HPßCD-IC is expected to be used as a green preservative for the storage and preservation of citrus fruits. PRACTICAL APPLICATION: In this study, LCEO was encapsulated in HPßCD by the saturated aqueous solution method and the prepared inclusion complex was characterized. The effects of LCEO/HPßCD-IC and LCEO/ßCD-IC on postharvest preservation of Shatang mandarin were compared. This work offers valuable insights into the postharvest preservation of citrus fruit by essential oil inclusion complexes.

Understanding the volatile organic compounds of 1-methylcyclopropylene fumigation and packaging on yellow-fleshed peach via headspace-gas chromatography-ion mobility spectrometry and chemometric analyses.

Headspace-gas chromatography-ion mobility spectrometry (HS-GC-IMS) and chemometric methods were utilized to analyze changes in volatile organic compounds (VOCs) of yellow-fleshed peach by 1-methylcyclopropylene (1-MCP) treatment and modified atmosphere packaging (MAP). Meanwhile, the storage quality of yellow-fleshed peach at room temperature (25°C) was also studied. Yellow-fleshed peach was treated by four methods, namely, nanomaterial packaging (NA), 1-MCP fumigation and nanomaterial packaging (1-MCP-NA), polyethylene (PE) packaging, and 1-MCP fumigation with polyethylene packaging (1-MCP-PE). Changes in the decay rate, firmness, browning index, soluble solid content, and titratable acid of the fruit were then measured at room temperature at 1, 4 and 10 days of storage. Thirty-two VOCs, including terpenoids, alcohols, esters, aldehydes, and ketones, were identified. Seventeen VOCs were found to be significant with predictive variable important in the projection (VIP) > 1 and p < 0.05 by analysis of variance and orthogonal projection to latent structure discriminant analysis (OPLS-DA). After 10 days of storage, the browning index of 1-MCP-NA group was 51.7%, which was lower than the control fruit (PE, 76.7%). Compared with other three treatments, 1-MCP-NA showed the better ability to delay and inhibit decreases in ester and aldehyde contents, and the ethanol content was lowest in the samples treated by 1-MCP-NA during storage. Differences among treatment groups were distinguished by principal component analysis (PCA) and hierarchical clustering heat map. The results showed that 1-MCP-NA could well maintain the quality and flavor stability of yellow-fleshed peach, and it had a good application prospect in the postharvest preservation of yellow-fleshed peach. PRACTICAL APPLICATION: In this study, 1-methylcyclopropylene (1-MCP) combined with nanomaterial (NA) packaging (1-MCP-NA) proved to have a better fresh preservation effect. 1-MCP-NA showed better ability to delay and inhibit decreases in ester and aldehyde contents by HS-GC-IMS technique. It provided a new strategy for postharvest storage of yellow-fleshed peaches.

Risk factors for urinary tract infection in kidney transplantation from brain death donor and its role in graft function. / è„‘æ­»äº¡å™¨å®˜æçŒ®è‚¾ç§»æ¤æœ¯åŽå‘ç”Ÿå°¿è·¯æ„ŸæŸ“çš„å±é™©å› ç´ åŠå ¶å¯¹ç§»æ¤è‚¾åŠŸèƒ½çš„å½±å“.

OBJECTIVES: Urinary tract infection (UTI) is the most common infection complication after kidney transplantation, and the reports of the incidence vary greatly among different centers. This study aims to explore the risk factors for UTI after kidney transplantation with the donation from brain death (DBD) and the impact on graft function, thus to provide theoretical basis for comprehensive prevention and treatment of UTI after kidney transplantation. METHODS: The clinical and laboratory data of DBD kidney transplantation from January 2017 to December 2018 in Xiangya Hospital, Central South University were collected and retrospectively analyzed. Patients were assigned into an UTI group and a non-UTI group. The base line characteristics, post-transplant complications, and graft function were compared between the 2 groups. Multivariate logistic regression was used to analyze the risk factors for UTI. RESULTS: A total of 212 DBD kidney transplant recipients were enrolled in this study. UTI occurred in 44 (20.75%) patients after transplantation. The female, the time of indwelling catheter, and postoperative urinary fistula were independent risk factors for UTI after DBD kidney transplantation. A total of 19 strains of gram-positive bacteria, 12 strains of gram-negative bacteria , and 10 strains of fungi were isolated from the urine of 44 UTI patients. The UTI after kidney transplantation significantly increased time of hospital stay (P<0.001) and raised the cost for antibiotics (P=0.004). The graft function was much worse in the UTI group compared with the non-UTI group (P<0.001) at 3 months after transplantation. Twenty (45.45%) patients recurred UTI within one year after transplantation. Non-hemodialysis before transplantation and perioperative combination of antibacterial and antifungal drugs were independent risk factors for recurrence of UTI. CONCLUSIONS: UTI after DBD kidney transplantation transplantation affects the renal function at 3 months and increases the patient's economic burden.

Detection of toxic methylenecyclopropylglycine and hypoglycin A in litchi aril of three Chinese cultivars.

As a subtropical fruit with high commercial values, litchi is also a source of methylenecyclcopropylglycine (MCPG) and hypoglycin A (HGA), which could cause hypoglycemia and fatal encephalopathy in human. In this work, a quantitative method was developed well to detect MCPG and HGA present in litchi aril of different cultivars. Method validation was evaluated well by linearity, recovery, precision and sensitivity. Among three cultivars, 'Feizixiao' contained the highest toxin level with 0.60-0.83 mg kg-1 of MCPG and 10.66-14.46 mg kg-1 of HGA, followed by 'Huaizhi' with 0.08-0.12 mg kg-1 of MCPG and 0.63-1.54 mg kg-1 of HGA, and 'Nuomici' with 0.09-0.11 mg kg-1 of MCPG and 0.35-0.91 mg kg-1 of HGA. The toxin levels were highly associated with litchi cultivar and storage time. These findings can provide new knowledge to help to recommend the safe consumption of fresh litchi based on human health.

MicroRNA­27a­3p promotes epithelial­mesenchymal transition by targeting NOVA alternative splicing regulator 1 in gastric cancer.

NOVA alternative splicing regulator 1 (NOVA1) dysregulation has been detected in the gastric cancer microenvironment. Decreased NOVA1 expression has been linked to the progression and poor prognosis of gastric cancer; however, the role of NOVA1 in regulating epithelial­mesenchymal transition (EMT) remains unclear in this disease. Experimental evidence has shown that miR­27a­3p is a potential oncogene in gastric cancer. In the present study, we observed that miR­27a­3p expression was increased in gastric cancer and was inversely associated with overall survival. Overexpression of miR­27a­3p promoted EMT in AGS gastric cancer cells. Additionally, overexpression of miR­27a­3p inhibited NOVA1 expression, while silencing of NOVA1 promoted EMT in AGS cells. A total of 108 gastric cancer samples were examined for NOVA1 expression by immunohistochemistry. Decreased NOVA1 expression was linked to lymph node metastasis, tumor­node­metastasis stage and shorter overall survival. Therefore, these results indicated that NOVA1 could be a potential tumor suppressive gene and that miR­27a­3p promotes EMT by targeting NOVA1 in gastric cancer.

Effect of Ozone Treatment on Flavonoid Accumulation of Satsuma Mandarin (Citrus unshiu Marc.) during Ambient Storage.

This study aimed to compare the flavonoid accumulation between ozone-treated and untreated Satsuma mandarin (Citrusunshiu Marc.) fruits. The fruits exposed to gaseous ozone were found to have higher antioxidant activities and content of flavonoid during the storage period by ultra-high performance liquid chromatography (UPLC). To reveal the molecular regulation of flavonoid accumulation by ozone, chalcone synthase (CHS), chalcone isomerase (CHI), ß-1,3-glucanase (GLU), chitinase (CHT), phenylalanine ammonia-lyase (PAL), and peroxidase (POD) were identified and their expression was examined by quantitative real-time polymerase chain reaction (q-PCR). These results support the promising application of ozone treatment as a safe food preservation technique for controlling postharvest disease and extending shelf-life of harvested Satsuma mandarin.

Pericarp and seed of litchi and longan fruits: constituent, extraction, bioactive activity, and potential utilization.

Litchi (Litchi chinensis Sonn.) and longan (Dimocarpus longan Lour.) fruits have a succulent and white aril with a brown seed and are becoming popular worldwide. The two fruits have been used in traditional Chinese medicine as popular herbs in the treatment of neural pain, swelling, and cardiovascular disease. The pericarp and seed portions as the by-products of litchi and longan fruits are estimated to be approximately 30% of the dry weight of the whole fruit and are rich in bioactive constituents. In the recent years, many biological activities, such as tyrosinase inhibitory, antioxidant, anti-inflammatory, immunomodulatory, anti-glycated, and anti-cancer activities, as well as memory-increasing effects, have been reported for the litchi and longan pericarp and seed extracts, indicating a potentially significant contribution to human health. With the increasing production of litchi and longan fruits, enhanced utilization of the two fruit by-products for their inherent bioactive constituents in relation to pharmacological effects is urgently needed. This paper reviews the current advances in the extraction, processing, identification, and biological and pharmacological activities of constituents from litchi and longan by-products. Potential utilization of litchi and longan pericarps and seeds in relation to further research is also discussed.

Comparative volatile compounds and primary metabolites profiling of pitaya fruit peel after ozone treatment.

BACKGROUND: Ozone treatment can effectively inhibit fruit decay in many fruits during postharvest storage. However, little information is available for pitaya fruit. RESULTS: Ozone treatment significantly reduced the decay rate and induced the enzyme activities of peroxidase and polyphenol oxidase, and also reduced the levels of reactive oxygen species. In total, 103 metabolites were detected and changed the content after ozone treatment, including 54 primary metabolites and 49 aromatic compounds. After significance and importance analysis, 37 metabolites were important. Some metabolites were induced by peel senescence to respond to senescence stress, including d-fructose, d-glucose, mannose, inositol, galactonic acid, ethanedioic acid and stearic acid. Some metabolic products of peel senescence were reduced by ozone treatment, including d-arabinose, glucaric acid, galacturonic acid, 1-hexanol, 4-ethylcyclohexanol, ß-linalool, palmitoleic acid and 2-hydroxy-cyclopentadecanone. Some metabolites induced by ozone treatment might play a vital role in delaying the senescence and decay, including malic acid, succinic acid, pentenoic acid, eicosanoic acid, 2-hexenal, hexanal, 2-heptenal, 4-heptenal, 2-octenal and nitro m-xylene. CONCLUSION: Ozone treatment significantly reduced decay and prolonged shelf-life without reducing fruit quality. In total, 37 metabolites might play an important role in ozone delayed fruit decay. © 2018 Society of Chemical Industry.

Essential role of microRNA-650 in the regulation of B-cell CLL/lymphoma 11B gene expression following transplantation: A novel mechanism behind the acute rejection of renal allografts.

Kidney transplantation is an effective final therapeutic procedure for patients with end-stage kidney failure. Although advanced immunosuppressive therapy is administered following transplantation, certain patients still suffer from acute allograft rejection. MicroRNAs (miRs) have a potential diagnostic and therapeutic value for acute renal allograft rejection; however, their underlying mechanism of action is largely unknown. In the present study, an increased level of miR-650 was identified to be associated with the downregulation of B-cell CLL/lymphoma 11B (BCL11B) expression in acute renal allograft rejection. Furthermore, in vitro study using human renal glomerular endothelial cells (HRGECs) transfected with a miR-650 mimic revealed that key characteristics of acute renal allograft rejection were observed, including apoptosis, the release of cytokines and the chemotaxis of macrophages, while the effects were reduced in HRGECs transfected with a miR-650 inhibitor. The existence of a conserved miR-650 binding site on the 3'-untranslated region of BCL11B mRNA was predicted by computational algorithms and confirmed by a luciferase reporter assay. Knockdown of BCL11B with small interfering RNA (siRNA) significantly increased the apoptotic rate and significantly decreased the proliferation ability of HRGECs compared with the negative control group. HRGECs transfected with a combination of BCL11B siRNA and the miR-650 mimic demonstrated a significant increase in the rate of apoptosis compared with the control. These results suggest that the upregulation of miR-650 contributes to the development of acute renal allograft rejection by suppression of BCL11B, which leads to apoptosis and inflammatory responses. Thus, miR-650 and BCL11B may represent potential therapeutic targets for the prevention of acute renal allograft rejection.

Comparative Transcriptome Analysis of Penicillium citrinum Cultured with Different Carbon Sources Identifies Genes Involved in Citrinin Biosynthesis.

Citrinin is a toxic secondary metabolite of Penicillium citrinum and its contamination in many food items has been widely reported. However, research on the citrinin biosynthesis pathway and its regulation mechanism in P. citrinum is rarely reported. In this study, we investigated the effect of different carbon sources on citrinin production by P. citrinum and used transcriptome analysis to study the underlying molecular mechanism. Our results indicated that glucose, used as the sole carbon source, could significantly promote citrinin production by P. citrinum in Czapek's broth medium compared with sucrose. A total of 19,967 unigenes were annotated by BLAST in Nr, Nt, Swiss-Prot and Kyoto Encyclopedia of Genes and Genomes (KEGG) databases. Transcriptome comparison between P. citrinum cultured with sucrose and glucose revealed 1085 differentially expressed unigenes. Among them, 610 were upregulated while 475 were downregulated under glucose as compared to sucrose. KEGG pathway and Gene ontology (GO) analysis indicated that many metabolic processes (e.g., carbohydrate, secondary metabolism, fatty acid and amino acid metabolism) were affected, and potentially interesting genes that encoded putative components of signal transduction, stress response and transcription factor were identified. These genes obviously had important impacts on their regulation in citrinin biosynthesis, which provides a better understanding of the molecular mechanism of citrinin biosynthesis by P. citrinum.

[Treatment of ureteral fistula and stricture following renal transplantation with Boari flap].

OBJECTIVE: To estimate the incidence rate of ureteral fistula and stricture after kidney transplantation, and to evaluate the effect of bladder flap (Boari flap) on ureteral complication of the transplanted kidney. 
 Methods: The clinical data and risk factors from 270 recipients of renal transplantation, who came from the Centre of Organ Transplantation, Xiangya Hospital, Central South University from January 2010 to January 2015, were retrospectively analyzed. The surgical management included Boari flap for ureteral reconstruction, neoureterocystostomy and endoscopic therapy with double-J (DJ) stent placement. Surgical proceeding and the effectiveness were evaluated.
 Results: The incidence rate of ureteral fistula following renal transplantation was 3.3%. The risk factors for ureteral fistula included elder donor age (P<0.05), delayed graft function (P<0.01), bladder spasm (P<0.05), and multiple renal arteries in allograft (P<0.01). Four cases were recovered after conservative treatment, and the other 5 cases were recovered after the treatment with Boari flap for ureteric reconstruction. The incidence rate of ureteral stricture was 4.4%. The risk factors for ureteral stricture included elder donor age (P<0.05), delayed graft function (P<0.05), cystospasm (P<0.05), ureteral fistula (P<0.01) and multiple renal arteries in allograft (P<0.01). Four cases underwent endoscopic therapy, 2 of them carried out percutaneous nephrostomy followed by antegrade DJ stent placement and the other 2 patients by retrograde DJ stent placement under ureteroscopy. Eight patients underwent surgery, 6 of them was treated by Boari flap for ureteral reconstruction and 2 patients were treated by neoureterocystostomy. All the patients recovered after surgical management.
 Conclusion: The ureteral complications after renal transplantation include ureteral fistula and stricture. Although the total incidence is low, the complications can result in adverse effects to the graft function and the life quality of the recipients. The risk factors for ureteral complication include elder donor age, delayed graft function, cystospasm, and multiple renal arteries in allograft. Ureteral fistula is the risk factor for ureteral fracture. Boari flap for ureterial reconstruction is an effective method in the treatment of the ureteral fistula and stricture.

Structural, Infrared and Magnetic Properties of Nanosized Ni(x)Zn1-xFe2O4 Powders Synthesized by Sol-Gel Technique.

Ni-Zn ferrites Ni(x)Zn1-xFe2O4 (x = 0.2, 0.4, 0.5, 0.6, 0.8) powders were synthesized by sol-gel technique. Structural, infrared and magnetic properties of samples were investigated. Spinel structural characteristics are shown by XRD spectra and the morphologies observed by atomic force microscopy demonstrate the samples are in nano-range. For all the samples, FTIR spectra exhibit obvious v1 infrared absorbing bands, in the range 500-600 cm-1, corresponding to intrinsic stretching vibrations of the metal ions at the tetrahedral site (Td), Mtetra <--> O. Furthermore, the central position of v1 band is tending to shift to larger wave numbers with the increasing Ni contents in the samples. For the samples Ni(x)Zn1-xFe2O4 (x = 0.2, 0.4), the v2 infrared absorbing bands, in the range 450-385 cm(-1), corresponding to stretching vibrations of the metal ions at the octahedral-metal stretching (Oh), Mocta <--> O, were also observed. However, for samples Ni(x)Zn1-xFe2O4 with higher Ni content (x = 0.5, 0.6, 0.8), the v2 infrared absorbing bands were obscure. The magnetic hysteretic loops at room temperature obtained from vibration samples magnetometer reveal the soft magnetism of the samples. The sample with lowest Ni content, Ni0.2Zn0.8Fe2O4, presents much higher saturation field than the other samples. The coercive field rises with increased Ni content, which is ascribed to the increased magnetocrystalline anisotropy constant with Ni content.

Effects of Ni Deposition on the Electrochemical Properties of CNT/Ni Electrode and Its Application for Glucose Sensing.

A low density CNT forest was fabricated by plasma enhanced chemical vapor deposition, and Ni nanoclusters were well distributed on the sidewall and on top of CNT forest by magnetron sputtering. The Ni deposition time plays an important role in electrochemical properties of the CNT/Ni electrodes, and the optimized deposition time is 150 to 240 s. Cyclic voltammetry and chronoamperometry were used to evaluate the catalytic activities of the CNT/Ni electrodes. The sensitivity of the glucose sensor based on a Ni24OS electrode is able to reach 1433 µA mM(-1) cm(-2), which is much higher than that found using a NiOS electrode.

Facile Preparation and Self-Assembly of Monodisperse Polystyrene Nanospheres for Photonic Crystals.

We have developed boiling emulsifier-free emulsion polymerization recipes for the synthesis of monodisperse polystyrene (PS) nanospheres with diameters between ca. 100 and 300 nm. The morphologies of the nanospheres during growth were characterized and the results showed that the PS nanospheres with uniform structures could be synthesized rapidly by modifying the reaction conditions. These nanospheres readily self-assemble into three-dimensionally colloidal photonic crystal film and whose photonic band-stop could be tuned over the entire visible spectral region by altering the sphere diameters.

[Transient stopping immunosuppressive agents during the post-transplant pulmonary infection does not affect the long-term outcome of renal transplantation].

OBJECTIVE: To determine the effect of transient withdrawal of immunosuppressive agents during the treatment of pulmonary infection on long-term survival of patients and graft s. METHODS: A total of 104 patients with post-transplant pulmonary infection were enrolled in this study. These patients received renal transplantation in Center for Organ Transplantation, Xiangya Hospital, Central South University, during December 2005 and August 2014. Among them, 50 patients stopped immunosuppressive agents during the treatment of infection. These patients served as stopping drug (SD) group, whereas the remaining patients who served as a control group did not stop immunosuppressive drugs. The five-year cumulative patient survival, graft survival, and laboratory results were compared between the 2 groups. RESULTS: The five-year cumulative patient survival rates in the SD group were significantly lower than those in the control group [(69.8 ± 7.0)% vs (94.2 ± 3.2)%, P=0.001]. There was no significant difference in the allograft survival rates between the 2 groups [(81.7 ± 6.6)% vs (90.9 ± 4.3)%, P=0.113]. In patients who survived from pulmonary infection, there was no significant difference in long-term survival rates between the 2 groups (P=0.979). CONCLUSION: Pulmonary infection impacts allograft survival after patients underwent renal transplantation. Transient stopping immunosuppressive agents during the treatment of infection is a safe and necessary treatment strategy for patients with serious post-transplant pulmonary infection.

shRNA-mediated GSTP1 gene silencing enhances androgen-independent cell line DU145 chemosensitivity.

PURPOSE: Design short hairpin RNA (shRNA) interference sequence to silence glutathione S-transferase P1 (GSTP1) gene of androgen-independent prostate cancer cell line DU145, and then to explore its effect on sensitivity to chemotherapeutics. METHODS: Target sequence was picked up to form the shRNA. DU145 cell was divided into five groups according to the shRNA added for transfection: shRNA255, shRNA554, shRNA593, negative-shRNA and blank group. Fluorescence microscope was used to pick up the shRNA with the highest transfection ratio. Western blotting and RT-PCR were taken to pick up the shRNA with the best gene silencing result. 3-(4,5-Dimethylthiazol-2-yl)-2, 5-diphenyl tetrazolium bromide assay and terminal de-oxynucleotidyl transferase-mediated dUTP nick end-labeling assay were used to detect survival ratio and apoptosis ratio of DU145 administered of fluorouracil (5-FU) or paclitaxel (PA) at different concentrations before and after shRNA transfection. RESULTS: Three different shRNA oligonucleotides (shRNA255; shRNA554; shRNA593) targeting the coding sequence of GSTP1 mRNA and one negative control shRNA were constructed. The transfection ratio of shRNA554 (76.2 ± 0.68 %) was higher than that of shRNA255 (63.3 ± 1.04 %) (P < 0.01) or shRNA593 (72.7 ± 0.33 %) (P < 0.01). After transfection of shRNA554, the mRNA and protein of level were the lowest, P < 0.01. The survival ratio of DU145 administered with 5-FU of different concentrations (30, 60, 120, 240 µg/ml) declined after transfection (P < 0.01). Besides, the apoptosis ratio increased after transfection (P < 0.01). Similarly the survival ratio of DU145 administered with PA of different concentrations (0.2, 2, 10, 20 µg/ml) declined (P < 0.01) and the apoptosis ratio increased (P < 0.01) after transfection. CONCLUSIONS: The gene GSTP1 silence via shRNA transfection to androgen-independent prostate cancer cell line DU145 enhances the sensitivity to chemotherapeutics.

[Effect of gene GSTP1 silencing via shRNA transfection on androgen independent prostate cancer cell line Du145].

OBJECTIVE: To design short hairpin RNA (shRNA) interference sequence to silence glutathione S-transferase P1 (GSTP1) gene of androgen independent prostate cancer cell line DU145, and to explore its effect on proliferation and sensitivity to chemotherapeutics. METHODS: The target sequence was picked up to form the shRNA, and the 3 shRNA expression vectors were shRNA255, shRNA554 and shRNA593. The DNA template was cloned to plasmid pGPU6/GFP/Neo. The shRNA was identified by enzyme digesting and gene sequencing. The screening experiment was done to pick up the shRNA expression vector with the highest transfection ratio and best gene silencing results. DU145 cells were divided into a blank plasmid group and a shRNA transfected group. According to the chemotherapeutics the DU145 cells were divided into a fluorouracil (FU) group and a paclitaxel (PA) group, and the 2 groups were subdivided into 4 subsets according to the chemotherapeutic concentrations (FU: 30, 60, 120, and 240 µg/mL; PA: 0.2, 2, 10, and 20 µg/mL), meanwhile a blank control group was included respectively. 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay was used to evaluate the proliferation after the transfection. MTT and terminal de-oxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL) assay were used to detect the inhibition effect of different concentrations of 5-FU or PA on the proliferation and induction of apoptosis of DU145. RESULTS: The transfection ratio of the 3 shRNA expression vectors (shRNA255, shRNA554, and shRNA593) was (63.30±1.04)%, (76.20±0.68)%, and (72.70±0.33)%, and the transfection ratio of shRNA554 was the highest. there was significant difference among the above 3 shRNA expression vectors (P<0.01). After the transfection, the mRNA was 128.31±2.50, 43.24±4.30 and 85.62±6.30, the GSTP1 protein was 163.92±12.40, 65.38±9.30 and 114.25±16.70. After the transfection of shRNA554, the mRNA and protein of GSTP1 were the lowest level. there was significant difference among the above 3 shRNA expression vector (P<0.01). MTT analysis showed that before the transfection, the survival ratio of cells under different concentrations of FU (30, 60, 120, and 240 µg/mL) was (95.60±2.11)%, (90.20±0.86)%, (83.10±3.12)% and (74.60±1.32)%; however after the transfection, the survival ratio of cells was (91.30±1.43)%, (84.60±2.13)%, (73.20±1.52)%, and (65.5±0.942)%. TUNEL assay showed that before the transfection, the apoptosis ratio of cells under different concentrations of FU (30, 60, 120, and 240 µg/mL) was (5.50±0.88)%, (10.20±1.64)%, (15.20±2.39)%, and (25.10±2.59)%; however after the transfection, the apoptosis ratio of cells was (10.8±0.62)%, (15.7±1.32)%, (20.4±1.89)%, and (34.9±2.54)%. After the transfection, the cell survival ratio decreased under the same concentration of FU, and the apoptosis ratio increased, with statistical significance (both P<0.01). MTT analysis showed that before the transfection, the survival ratio of cells under different concentrations of PA (0.2, 2, 10, and 20 µg/mL) was (98.50±2.34)%, (95.20±1.32)%, (89.40±0.68)%, and (82.70±1.73)%; after the transfection the survival ratio of cells was (94.20±0.78)%, (86.50±2.13)%, (78.70±1.34)%, and (70.10±0.76)%. TUNEL assay showed that before the transfection, the apoptosis ratio of cells under different concentrations of PA (0.2, 2, 10, and 20 µg/mL) were (2.40±1.07)%, (5.20±1.33)%, (10.50±2.41)%, (20.70±1.92)%; after the transfection the apoptosis ratio of cells was (5.46±2.13)%, (13.80±1.24)%, (21.20±2.39)%, and (29.20±2.21)%. After the transfection, the cell survival ratio decreased under the same PA concentration, and the apoptosis ratio increased, with statistical significance (both P<0.01). CONCLUSION: gene GSTP1 silence via shRNA transfection to androgen independent prostate cancer cell line DU145 can inhibit its proliferation in time dependent manner, and induce apoptosis and raise its sensitivity to chemotherapeutics.

Analysis of the relationship between postoperative ophthalmic complications and dialysis time of pre-kidney transplantation.

AIM: To determine the influence of the dialysis time before kidney transplantation on postoperative ophthalmic complications. METHODS: One hundred and eighty three patients who were given the follow-up after kidney transplantation were selected, including 124 males and 59 females. The dialysis time before kidney transplantation was (2.9±2.1) years. Among them, there were 93 cases having cadaveric renal transplantation and 90 cases having living relative renal transplantation. The conditions of ophthalmic complications in all the patients after kidney transplantation were investigated and the incidence rate on ophthalmic complications having different dialysis time before kidney transplantation was given Chi-square test and Chi-square linear trend test. RESULTS: Among 183 patients with kidney transplantation, 95 patients (51.9%) had at least one ophthalmic complication and the rest 88 patients (48.1%) had no significant abnormality at the eye region. The most common ophthalmic complications were pinguecula/conjunctival degeneration (31 cases), the following was caligo lentis (24 cases). The main manifestations were grayish white granule and plaque turbidity occurred in posterior capsule at the posterior pole of crystaline lens. The angulus iridocornealis of 5 patients (5.3%) with cataract and glaucoma were all open-angle through the detection by gonioscope. Through visual field examination, there were 2 patients with paracentral scotoma, 2 patients with arcuate scotoma and one case with nasal step. CONCLUSION: The experiments verify that the incidence of glaucomawas relates to the dialysis time before kidney transplantation, and the incidence rate might be higher if the dialysis time is longer.