Mammography-based radiomics analysis and imaging features for predicting the malignant risk of phyllodes tumours of the breast.

AIM: To determine whether the mammography (MG)-based radiomics analysis and MG/ultrasound (US) imaging features could predict the malignant risk of phyllodes tumours (PTs) of the breast. MATERIALS AND METHODS: Seventy-five patients with PTs were included retrospectively (39 with benign PTs, 36 with borderline/malignant PTs) and divided into thetraining (n=52) and validation groups (n=23). The clinical information, MG and US imaging characteristics, and histogram features were extracted from craniocaudal (CC) and mediolateral oblique (MLO) images. The lesion region of interest (ROI) and perilesional ROI were delineated. Multivariate logistic regression analysis was performed to determine the malignant factors of PTs. Receiver operating characteristic (ROC) curves were generated, and the area under the curve (AUC), sensitivity, and specificity were calculated. RESULTS: There was no significant difference found in the clinical or MG/US features between benign and borderline/malignant PTs. In the lesion ROI, variance in the CC view and mean and variance in the MLO view were independent predictors. The AUC was 0.942, sensitivity and specificity were 96.3% and 92%, respectively, in the training group. In the validation group, the AUC was 0.879, the sensitivity was 91.7%, and the specificity was 81.8%. In the perilesional ROI, the AUCs were 0.904 and 0.939, sensitivities were 88.9% and 91.7%, and the specificities were 92% and 90.9% in the training and validation groups, respectively. CONCLUSIONS: MG-based radiomic features could predict the risk of malignancy of patients with PTs and may be used as a potential tool to differentiate benign and borderline/malignant PTs.

[Value of (18)F-FDG PET-CT imaging to predict epidermal growth factor receptor mutations in patients with lung squamous cell carcinoma].

Objective: To investigate the value of (18)F-fluorodeoxy glucose ((18)F-FDG) positron emission tomography/computed tomography (PET-CT) in predicting the epidermal growth factor receptor (EGFR) mutations in patients with lung squamous cell carcinoma. Methods: We retrospectively analyzed the clinical data and (18)F-FDG PET-CT imaging data of 206 patients with lung squamous cell carcinoma confirmed by pathology and underwent EGFR mutation test in the First Affiliated Hospital of Nanjing Medical University from June 2013 to October 2018. Receiver operating characteristic (ROC) curve analysis was performed to quantify the predictive value of maximum standard uptake value (SUV(max)), metabolic tumor volume (MTV), total lesion glycolysis (TLG). The Chi-squared test was used to assess the difference in PET parameters. A multivariate Logistic regression analysis was performed to yield the parameters with statistic difference. Results: All of 206 patients with lung squamous cell carcinoma showed a high (18)F-FDG uptake. The median of SUV(max), MTV and TLG were 19.14, 37.69 cm(3) and 291.73, respectively. Among the 206 patients, EGFR mutations were identified in 14 cases, including 7 with exon 21 (L858R) mutation, 6 with exon 19 mutation and 1 with exon 20 mutation. ROC curve showed that the AUC of SUV(max), MTV and TLG were 0.624 (95% CI=0.454-0.794, P=0.122), 0.892 (95% CI=0.811-0.973, P<0.001) and 0.860 (95% CI=0.768-0.952, P<0.001), respectively. The median SUV(max) (19.14) was used as the cutoff points due to the small value of AUC. The cutoff point of MTV was 20.09 cm(3), the cutoff point of TLG was 211.07. Univariate analysis showed that the sex, smoking history, M stage, MTV and TLG were associated with EGFR mutations (all P<0.05). Logistic multivariate analysis showed that the sex, smoking history and TLG were the independent predictors of EGFR mutation (all P<0.05). Conclusion: TLG detected by (18)F-FDG PET/CT is an independent factor for predicting EGFR mutation in patients with lung squamous cell carcinoma, and has certain reference value for predicting EGFR mutation.

[Macular buckling for highly myopic traction maculopathy].

Objective: To observe the efficacy of macular buckling in the treatment of highly myopic traction maculopathy. Methods: Retrospective case series study. The patients with high myopia who underwent macular buckling at the Zhongshan Ophthalmic Center of Sun Yat-sen University from June 2014 to June 2019 were enrolled, including 136 males and 212 females. The age was (56.68±11.59) years old. The outcomes measured included retinal reattachment rate, foveoschisis recovery rate, macular hole closure rate, postoperative best corrected visual acuity (BCVA), axial length (AL), and complications. The measurements were recorded preoperatively and at 1 month, 6 months, 1 year, 2 years, and 3 years postoperatively. The data was statistically analyzed using paired t test. Results: A total of 378 eyes were included, including 216 eyes with foveoschisis and macular detachment and 162 eyes with macular holes and macular detachment. Among them, 296 eyes underwent macular buckling, and the other 82 eyes underwent macular buckling combined with pars plana vitrectomy. During the follow-up period, 373 eyes (98.68%) achieved retinal reattachment; in patients with foveoschisis, 204 eyes (94.44%) were recovered; in patients with macular holes, 89 eyes (54.09%) achieved closure. All the postoperative results of BCVA were better than the preoperative value (1.459±0.841). BCVA continued to increase from postoperative month 1, remained stable at 1 year, and reached 0.908±0.606 at 3 years (t=6.896, P<0.01). All the postoperative results of AL were shorter than the preoperative value. The AL shortened by (4.423±1.740)mm at one month (t=33.144, P<0.01), increased gradually thereafter, remained stable at 1 year, and shortened by (2.101±1.643) mm at three years (t=6.392, P<0.01). The common complications included transient high intraocular pressure in 98 eyes (25.92%), epiretinal hemorrhage in 67 eyes (17.72%), and vitreous hemorrhage in 9 eyes (2.38%), which all resolved spontaneously within 1 month. In the early postoperative period, all patients had a certain degree of eye movement limitation, and 39 eyes (10.31%) had diplopia which resolved within 6 months without treatment. The strabismus surgery was arranged to treat esotropia in 6 eyes (1.58%). The macular buckle was removed from 1 eye (0.26%) because of the inability to tolerate diplopia. There were 8 eyes (2.11%) requiring a second operation to adjust the position of the buckle. The macular buckle was also removed from 4 eyes (1.05%) due to the implant rejection. Conclusion: Macular buckling can effectively shorten the AL, resolve posterior scleral staphyloma, and improve vision in the treatment of highly myopic traction maculopathy. (Chin J Ophthalmol, 2021, 57: 433-439).

Corticosteroid therapy for patients with severe novel Coronavirus disease 2019.

OBJECTIVE: To investigate the effect of corticosteroid on hospital mortality, hospital length of stay, and time of viral clearance in patients with severe and critical COVID-19. PATIENTS AND METHODS: Patients with severe and critical COVID-19 who had been discharged or expired were enrolled in this study. Patients were divided into corticosteroid group and non-corticosteroid group according to the systemic corticosteroid use or not. Clinical data were collected, and hospital mortality, hospital length of stay, time of viral clearance, time of mechanical ventilation, and duration from illness onset to symptom resolution were compared between the two groups. RESULTS: A total of 72 inpatients who were diagnosed with severe and critical COVID-19 were enrolled, in which 47 patients were divided into corticosteroid group and 25 were involved as the non-corticosteroid group. Baseline characteristics were generally similar between the two groups. Four (5.6%) patients died during hospitalization, and 68 (94.4%) were discharged. Among survivors, the mean duration time from admission to discharge was 19.5d (SD 7.05 d). The mean time of viral clearance among survivors was 17.5d (SD 7.67 d), with a maximum of 37 d, and a minimum of 5 d. Hospital mortality (4.3% vs. 8.0%), length of hospital stay (18.7d vs. 21.0d), and time of viral clearance (16.1d vs. 19.4d) had no significant difference between two groups (p>0.05). The duration of symptoms suffering was shorter in the corticosteroid group than non-corticosteroid group, with statistically significant difference (p<0.05). CONCLUSIONS: Corticosteroid therapy in patients with severe COVID-19 cannot reduce the hospital mortality, and is not associated with delayed viral clearance, but it could relieve the inflammatory storm and improve clinical symptoms in brief. Patients with severe COVID-19 could benefit from low-dose corticosteroid treatment.

[Reference value of urine tangible component analyzer for healthy adults in Guangzhou area].

Objective: To establish the normal reference for iQ200 urine component analyzer of healthy adults in Guangzhou area. Methods: Five hundreds and four normal urine samples were screened from the healthy population with the normal dry chemistry results of urine and normal biochemical results of plasma, and the RBC, WBC and epithelial cells were determined by iQ200 urine component analyzer. Then 1 ml urine was accurately collected from the samples by centrifugation and the RBC, WBC and epithelial cells were counted with neubauer counter by two inspectors. The samples were divided into 4 groups according to the age (18-45 years and >45 years old) and sex (male and female). The counts of RBC, WBC and epithelial cell of 4 groups with iQ200 urine component analyzer and neubauer counter were statistically analyzed. Results: The difference was not significant in RBC, WBC and epithelial cells between the results of iQ200 urine component analyzer and the results of neubauer counter(all P>0.05). There was not significant difference of RBC, WBC and epithelial cells of the same sex and different age groups (all P>0.05). There was significant difference of RBC, WBC and epithelial cells between the different gender groups(all P<0.05). The 95% confidence interval reference range of iQ200 urine component analyzer were as follows: RBC 0-7 cells/µl (male), 0-8 cells/µl (female); WBC 1-6 cells/µl (male), 1-8 cells/µl (female); epithelial cells 0-2 cells/µl (male), 0-8 cells/µl (female). Conclusion: The reference of this research of iQ200 urine component analyzer will be useful for the laboratory and clinic, but still need the verification experiment before using in the laboratory.

[Study of mumps immunity after administrating measles-mumps-rubella vaccine among children aged 2-7 years old in Jiangsu Province in 2015].

Objective: To investigate the immunity to mumps after administrating measles-mumps-rubella vaccine (MMR) among children aged 2-7 years old in Jiangsu province in 2015. Methods: A total of 4 190 healthy children aged 2-7 years old, living in local places for at least 3 months, and having been vaccinated at least 1 dose MMR were recruited to the study from Wujin district of Changzhou city, Gaogang district of Taizhou city and Ganyu district of Lianyungang city by using stratified cluster random sampling method between September and November, 2015. Those who did not accept MMR vaccination, who refused venous blood collection, who had affected mumps according to the memory of parents or teachers and who were diagnosed serious disease by clinical doctors were excluded from study. The self-designed questionnaire was used to collect the general information of the subjects and their MMR immunization history; and 0.5-2.0 ml of venous blood was collected from each subject. ELISA was used to detect the mumps antibody level in the serum of patients. Positive was defined as the antibody level ≥108 mU/ml, and negative as <108 mU/ml. χ(2) test was used to compare the difference in positive rates among subjects; and analysis of variance was used to compare the GMC changes in different time points after MMR vaccination. Results: Among 4 190 children, 2 280 were males (54.42%) and 1 910 were females(45.58%), and the positive rate of IgG antibody was 81.38% (3 344). There were 3 156 (95.18%) children vaccinated with one dose MMR, 187 (4.80%) children with two dose MMR, and 1 (0.02%) child with three dose MMR. The difference in positive rate of IgG antibody among different aged subjects showed statistical significance (χ(2)=58.61, P<0.001), the highest positive rate was in group of subjects aged 4-5 years old, at 89.43% (406/454), while the lowest positive rate was found among subjects aged 6-7 years old, at 75.63% (1 648/2 179). The positive rate after one dose of MMR vaccination was 79.14% (3 156/3 988), significantly less than it after two doses (93.03%, 187/201) (χ(2)=22.93, P<0.001). The GMC level at years<1, 1-<2, 2-<3, 3-<4, ≥4 following one dose MMR in the 3 988 children was 152.47, 227.78, 167.08, 126.91, 79.43 mU/ml, whose difference was statistically significant (F=51.29, P<0.001). Conclusion: The sero-prevalence of IgG antibody in the children aged 2-7 years old in Jiangsu province was high. The positive rate among who received two doses MMR was significantly higher than it among who received just one dose, and the GMC level waned with times.

Activated human hydroxy-carboxylic acid receptor-3 signals to MAP kinase cascades via the PLC-dependent PKC and MMP-mediated EGFR pathways.

BACKGROUND AND PURPOSE 3-Hydroxy-octanoate, recently identified as a ligand for, the orphan GPCR, HCA(3), is of particular interest given its ability to treat lipid disorders and atherosclerosis. Here we demonstrate the pathway of HCA(3)-mediated activation of ERK1/2. EXPERIMENTAL APPROACH Using CHO-K1 cells stably expressing HCA(3) receptors and A431 cells, a human epidermoid cell line with high levels of endogenous expression of functional HCA(3) receptors, HCA(3)-mediated activation of ERK1/2 was measured by Western blot. KEY RESULTS HCA(3)-mediated activation of ERK1/2 was rapid, peaking at 5 min, and was Pertussis toxin sensitive. Our data, obtained by time course analyses in combination with different kinase inhibitors, demonstrated that on agonist stimulation, HCA(3) receptors evoked ERK1/2 activation via two distinct pathways, the PLC/PKC pathway at early time points (≤ 2 min) and the MMP/ epidermal growth factor receptor (EGFR) transactivation pathway with a maximum response at 5 min. Furthermore, our present results also indicated that the ßγ-subunits of the G(i) protein play a critical role in HCA(3)-activated ERK1/2 phosphorylation, whereas ß-arrestins and Src were not required for ERK1/2 activation. CONCLUSIONS AND IMPLICATIONS We have described the molecular mechanisms underlying the coupling of human HCA(3) receptors to the ERK1/2 MAP kinase pathway in CHO-K1 and A431 cells, which implicate the G(i) protein-initiated, PLC/PKC -and platelet-derived growth factor receptor/EGFR transactivation-dependent pathways. These observations may provide new insights into the pharmacological effects and the physiological functions modulated by the HCA(3)-mediated activation of ERK1/2.

Pulmonary toxicity in mice exposed to low and medium doses of water-soluble multi-walled carbon nanotubes.

Carbon nanotubes (CNTs) are a class of new allotrope of carbon. Different functionalized CNTs may vary from their physical and chemical properties to the biological property. In this study, the toxicity of water-soluble taurine multi-walled CNTs (tau-MWNTs), raw MWNTs and positive control crystalline silicon dioxide particles on mouse lungs via intratracheal instillation (i.t.) was investigated. The dosages we used were 0.125, 0.25, 0.5 or 1 mg/kg of tau-MWNTs and raw MWNTs, and 1 mg/kg of silicon dioxide particles; Serum and lungs were collected at 1, 7, 14 or 28 days postexposure. The biochemical and cellular parameters were assessed, which include the ratio of the lung weight and body weight (lung indices), lactate dehydrogenase (LDH), alkaline phosphatase (ALP) and angiotensin converting enzyme (ACE) in serum, and malondialdehyde (MDA), reduced glutathione (GSH), total sulfhydryl group (TSH) in lung tissue homogenates as well as the hydroxyproline in lungs. The characteristic recovery of the lung injury at 28 days postexposure was examined by the assessment of LDH, ALP, lung indices, and histopathology. ACE, MDA, GSH, TSH and histopathological changes showed that tau-MWNTs were less toxic than the raw MWNTs. Histopathological and ultrastructural investigation indicated that the acute pulmonary inflammation in lungs alleviated after 7d postexposure, and were greatly recovered within 28d. Meanwhile, the entrapment of tau-MWNTs reduced greatly by the 28d postexposure. Whereas the heavier pathologic changes induced by raw MWNTs lasted 7 days more than that of tau-MWNTs. Notably, no occurrence of granulomas and fibrosis were found in this study both in the two CNTs samples through 28d postexposure. Silicon dioxide particles, on the contrary, produced more severe damage to lungs than CNTs did in lung index, as well as other biochemical and cellular parameters. These findings indicate that water-soluble tau-MWNTs in low and medium doses induce slight and recoverable pulmonary inflammation in mice, and are less toxic than the insoluble raw MWNTs.

Exploring rat plasmatic proteomes: what triggered the liver regeneration?

To further clarify the priming mechanism of liver regeneration, proteins and protein complexes from rat plasma (normal group, partial hepatectomy (PHx) group and sham-operation group) were comparatively studied by two-dimensional gel electrophoresis and two-dimensional blue native gel electrophoresis. Our results suggested that Kupffer cell--NF-kappaB/ROS might trigger the liver regeneration.

Diameter effects on cytotoxicity of multi-walled carbon nanotubes.

In this work, we focus on the diameter effects on cytotoxicity of different diametered multi-walled carbon nanotubes (MWNTs). Cellular viability, phagocytotic ability, and apoptosis of guinea pig alveolar macrophages (AM) exposed to MWNTs were studied. MWNTs in smaller diameter showed less cytotoxicity than the larger ones at the same dosage. The phagocytosis ability of AM was partially impaired when the concentration of MWNTs was up to 20 microg/ml, where a further characteristic feature of AM apoptosis was observed. The results indicate that MWNTs in different diameters exhibited quite different cytotoxicity, and for the first time revealed the diameter-effect of MWNTs on cytotoxicity.

Determination and pharmacokinetic study of indirubin in rat plasma by high-performance liquid chromatography.

A specific, simple and sensitive HPLC method with UV detection was developed and validated for the pharmacokinetic studies of indirubin in rat plasma for the first time. Indirubin, with osthole as the internal standard, was extracted from plasma samples by liquid-liquid extraction. Chromatographic separation was conducted on a reverse-phase ODS column (200 mm x 4.6 mm, i.d., 5 microm), using a mixture of methanol-water (75:25, v/v) as the mobile phase at a flow rate of 1.0 ml/min with UV detection at 289 nm. The calibration curve of indirubin was linear over the range of 6.5-1950 ng/ml in rat plasma. The lower limit of quantification (LLOQ) was found to be 6.5 ng/ml. The present method was successfully applied for estimating the pharmacokinetic parameters of indirubin following intravenous and intraperitoneal administration of indirubin to rats.

Bcl-xL transformed peanut (Arachis hypogaea L.) exhibits paraquat tolerance.

The human Bcl-xL gene was transformed into peanut cultivar Georgia Green via microprojectile bombardment. Following selection on hygromycin-containing medium and regeneration, eighty hygromycin-resistant callus clusters were recovered. Southern blot analysis of ten fertile lines revealed multiple insertions of the Bcl-xL transgene in most lines. Western blot analysis of primary plants and T1 progenies demonstrated detectable levels of Bcl-xL expression in four transgenic lines. We could not detect Bcl-xL protein in other tested lines even though transcripts were identified by RT-PCR and northern blot. Three of the western-positive transgenic lines either were sterile or the progenies lost the expressive copy of Bcl-xL. Only T1 progenies from line BX25-4-2a-19 continued to express an intermediate level of Bcl-xL. This line demonstrated paraquat tolerance at the 5 microM level. Tolerance to salt of T1 and T2 seeds from seven other transgenic lines also was tested, but no tolerance was found in these lines. A high level of Bcl-xL transgene expression may be deleterious to plant growth and development even though the gene may confer tolerance to other abiotic and biotic stresses such as drought and pathogens.

The adhesive properties of endothelial cells on endovascular stent coated by substrates of poly-l-lysine and fibronectin.

Optimizing endothelial cell growth and adhesion on the surface of metallic stents implanted in the vascular system is a fundamental issue in understanding and improving their long-term biocompatibility. The ability of the endothelial cell to attach and adhere to the luminal stent surface as well as the capacity to withstand the significant shear stress associated with blood flow are important determinants. The adhesive characteristics of human umbilical vein endothelial cellsectin (HUVEC) on stent surfaces coated with either Poly-L-Lysine (PLL) or fibron (FN) were compared with uncoated controls. Increasing concentrations of PLL and FN were measured using a micropipette aspiration system. The adhesivenamic properties of HUVECs under static flow conditions were compared to a dy environment on endovascular stents using a parallel-plate-flow chamber. A scanning electron microscope picture was used to measure the number and the adhesive cell ratio as well as the percentage of surface coverage of stent by endothelial cells. The adhesive forces of HUVECs on foreign surfaces coated with PLL and FN were higher compared to uncoated surfaces, and were dependent on incr ing concentrations. These coatings resulted in significant increase of the adhesive force of HUVECs. The influence of substrates on the adhesion of the endothelial cell monolayer under static or dynamic flow conditions was highly significant compared with controls (p<0.01). No significant differences were observed between PLL and FN substrates. Both PLL and FN coated surfaces can significantly increase the adhesion and growth of HUVECs on metallic stent surfaces.

PYY3-36 as an anti-obesity drug target.

The neuropeptide Y (NPY)/peptide YY (PYY) system has been implicated in the physiology of obesity for several decades. More recently ignited enormous interest in PYY3-36, an endogenous Y2-receptor agonist, as a promising anti-obesity compound. Despite this interest, there have been remarkably few subsequent reports reproducing or extending the initial findings, while at the same time studies finding no anti-obesity effects have surfaced. Out of 41 different rodent studies conducted (in 16 independent labs worldwide), 33 (83%) were unable to reproduce the reported effects and obtained no change or sometimes increased food intake, despite use of the same experimental conditions (i.e. adaptation protocols, routes of drug administration and doses, rodent strains, diets, drug vendors, light cycles, room temperatures). Among studies by authors in the original study, procedural caveats are reported under which positive effects may be obtained. Currently, data speak against a sustained decrease in food intake, body fat, or body weight gain following PYY3-36 administration and make the previously suggested role of the hypothalamic melanocortin system unlikely as is the existence of PYY deficiency in human obesity. We review the studies that are in the public domain which support or challenge PYY3-36 as a potential anti-obesity target.

Latent membrane protein 1 encoded by Epstein-Barr virus modulates directly and synchronously cyclin D1 and p16 by newly forming a c-Jun/Jun B heterodimer in nasopharyngeal carcinoma cell line.

Recently we confirmed that latent membrane protein 1 (LMP1) encoded by Epstein-Barr virus (EBV) accelerates a newly forming active c-Jun/Jun B heterodimer, a transcription factor, but little is known about the target gene regulated by it. In this paper, results indicated that a c-Jun/Jun B heterodimer induced by LMP1 upregulated cyclin D1 promoters activity and expression, on the contrary, downregulated p16, and maladjustment of cyclin D1 and p16 expression accelerated progression of cell cycle. Firstly, we found a c-Jun/Jun B heterodimer regulated synchronously and directly cyclin D1 and p16 in the Tet-on-LMP1-HNE2 cell line, in which LMP1 expression is regulated by Tet-on system. This paper investigated in depth function of the newly forming active c-Jun/Jun B heterodimer, and built new connection between environmental pathogenic factor, signal transduction and cell cycle.

Physiology: does gut hormone PYY3-36 decrease food intake in rodents?

Batterham et al. report that the gut peptide hormone PYY3-36 decreases food intake and body-weight gain in rodents, a discovery that has been heralded as potentially offering a new therapy for obesity. However, we have been unable to replicate their results. Although the reasons for this discrepancy remain undetermined, an effective anti-obesity drug ultimately must produce its effects across a range of situations. The fact that the findings of Batterham et al. cannot easily be replicated calls into question the potential value of an anti-obesity approach that is based on administration of PYY3-36.

Chronic ethanol consumption induces gene expression of pancreatic monitor peptide, but not SPINK1/PSTI-56, in rats.

The primary factors that predispose humans to the development of alcoholic pancreatitis are unknown. One of the earliest observations in humans in whom this disease develops is pancreatic hypersecretion caused by unknown mechanisms. Messenger RNA (mRNA) differential display was performed in a rat model to investigate the molecular mechanisms associated with ethanol-induced pancreatic hypersecretion. Male Wistar rats were pair-fed Lieber-DeCarli diets with or without ethanol for 7 days or 4 weeks. Total RNA was extracted from the pancreas and its neurohormonal control sites. Differentially expressed complementary DNA (cDNA) tags were isolated, cloned, and sequenced. One 248-bp cDNA was consistently and strongly induced in the pancreata of rats fed ethanol for 4 weeks. The sequence was highly homologous to both rat pancreatic monitor peptide (MP) and pancreatic secretory trypsin inhibitor (PSTI-56), also known as serine protease inhibitor, Kazal type 1 (SPINK1). Confirmatory reverse-transcription-polymerase chain reaction showed that PSTI-56 expression remained unchanged, whereas MP mRNA levels were elevated more than four times in the pancreata of ethanol-fed rats. These results indicate that long-term ethanol ingestion increases MP mRNA levels in the rat pancreas. Because MP stimulates cholecystokinin release and cholecystokinin is an important stimulant of pancreatic secretion, the enhanced MP gene expression may contribute to pancreatic hypersecretion.

Transgenic peanut plants obtained by particle bombardment via somatic embryogenesis regeneration system.

After pre-culture and treatment of osmosis, cotyledons of immature peanut (Arachis hypogaea L.) zygotic embryos were transformed via particle bombardment with a plasmid containing a chimeric hph gene conferring resistance to hygromycin and a chimeric intron-gus gene. Selection for hygromycin resistant calluses and somatic embryos was initiated at 10th d post-bombardment on medium containing 10-25 mg/L hygromycin. Under continuous selection, hygromycin resistant plantlets were regenerated from somatic embryos and were recovered from nearly 1.6% of the bombarded cotyledons. The presence and integration of foreign DNA in regenerated hygromycin resistant plants was confirmed by PCR (polymerase chain reaction) for the intron-gus gene and by Southern hybridization of the hph gene. GUS enzyme activity was detected in leaflets from transgenic plants but not from control, non-transformed plants. The production of transgenic plants are mainly based on a newly improved somatic embryogenesis regeneration system developed by us.

Rat mitochondrial ATP synthase ATP5G3: cloning and upregulation in pancreas after chronic ethanol feeding.

Individuals with chronic excessive alcohol ingestion are put at the risk of acute and chronic pancreatitis. Underlying molecular mechanisms are unknown. Differential gene expression in the pancreas was profiled using mRNA differential display by comparison between control and ethanol-consuming rats. Male Wistar rats were fed with diets containing 6.7% (vol/vol) ethanol for 4 wk. A cDNA tag that was overexpressed in the pancreas of rats fed ethanol was isolated. A 723-bp cDNA was cloned from a rat pancreatic cDNA library, which encodes a novel rat mitochondrial ATP synthase subunit 9, isoform 3 (ATP5G3), which is homologous to a human ATP5G3 gene. Real-time PCR demonstrated that all three nuclear gene isoforms (ATP5G1, ATP5G2, and ATP5G3) were consistently upregulated in the pancreas of alcohol-consuming rats, parallel with mitochondrial injury. The cellular response to mitochondrial damage and metabolic stress may reflect an adaptive process for mitochondrial repair in pancreatic acinar cells during chronic ethanol ingestion.