[Comparison of quantitative detection of BCR::ABL1 p210 transcript levels: a multicenter study].

Objective: To assess the capability of seven reference medical laboratories to detect BCR::ABL1 p210 transcription levels and to compare the results among those laboratories. Methods: The interlaboratory comparison was carried out in two stages. The samples were prepared by the reference laboratory. The quantitative values of BCR::ABL1 p210 of the comparison samples covered 0.001%-0.01%, 0.01%-0.1%, 0.1%-1%, 1%-10% and>10% in each stage. Real-time quantitative PCR (RT-PCR) and dPCR (digital PCR) were used to examine the samples. The conversion factor (CF) was calculated and validated for each laboratory. Results: In the RT-PCR comparison, one laboratory was failed to detect BCR::ABL1 p210 in fourteen samples at the first stage. The results of the other six laboratories were qualified with the bias <±1.2 folds (-0.133-0.338) and 95% limits of agreement within ±5 folds (upper limit 0.147-0.785, lower limit -0.770--0.109), and the corresponding CF values were calculated and validated. In the dPCR comparison, one laboratory did not report results at the second stage. The results of the other six laboratories were qualified with the bias <±1.2 folds (-0.026-0.267) and 95% limits of agreement within±5 folds (upper limit 0.084-0.991, lower limit -0.669--0.135), and the corresponding CF values were calculated and validated. The samples with BCR::ABL1 p210 quantitative values of 0.01%-0.1%, 0.1%-1%, 1%-10% and >10% could be detected by both RT-PCR and qPCR. When the quantitative value of BCR::ABL1 p210 was 0.001%-0.01%, the detection rate of dPCR was higher than that of RT-PCR (85.56% vs. 68.00%). Conclusions: A good consistency is present among various laboratories. The quantitative value of BCR::ABL1 p210 is comparable among laboratories as shown by the CF value conversion. For quantitative detection of BCR::ABL1 p210 deep molecular reaction, dPCR has a higher positive detection rate and more advantages than RT-PCR. To ensure the accuracy and reproducibility of the BCR::ABL1 p210 test, it is imperative for every laboratory to enhance their daily quality control practices.

Periodontitis and the risk of oral, gastric and esophageal cancers: a two-sample Mendelian randomization study.

BACKGROUND: Periodontitis is a common oral disease and the chronic inflammation caused by it may influence the development of cancers in the upper gastrointestinal tract. Many observational studies have established a relationship between the two, but the results are not entirely consistent. METHODS: Two-sample MR was performed using publicly available genome-wide association studies data for periodontitis, oral, gastric and oesophagal cancers. The Inverse Variance Weighting (IVW) method serves as the primary method, with MR Egger, Weighted Median, Simple Model and Weighted Model Algorithm methods as complementary methods to assess genetic causal associations. Cochran Q-test, MR-Egger regression and MR polytropic residuals and outliers were used to assess heterogeneity and horizontal pleiotropy. RESULTS: IVW results did not support a causal association between periodontitis and oral (OR = 1.00, 95% CI: 1.00, 1.00) and oesophagal cancer (OR = 1.00, 95% CI: 1.00, 1.00). Similarly, there was again no causal association between periodontitis and gastric cancer, which was integrated with an OR of 1.04 (95% CI: 0.97, 1.12). Complementary method results were consistent with IVW and heterogeneity and horizontal pleiotropy were not found in most studies. CONCLUSIONS: The findings of our MR study do not support a causal relationship between periodontitis and oral, gastric and oesophagal cancers.

[Clinical features of non-cirrhotic portal hypertension in patients with common variable immunodeficiency].

We wished to summarize the clinical features of common variable immunodeficiency (CVID) complicated by non-cirrhotic portal hypertension (NCPH) and to deepen our understanding of it. The case data of CVID complicated with NCPH admitted to Peking Union Medical College Hospital from January 1983 to May 2021 were analyzed retrospectively to summarize their clinical characteristics. Six patients with CVID combined with NCPH (three of each sex; 16-45 years) were assessed. Four patients had portal hypertension. All patients had anemia, splenomegaly, a normal serum level of albumin and transaminases, and possibly increased levels of alkaline phosphatase and gamma-glutamyl transpeptidase. Two patients were diagnosed with esophagogastric fundic varices by gastroscopy. Two patients underwent splenectomy (which improved hematologic abnormalities partially). Four patients had autoimmune disease. Two cases were diagnosed with nodular regenerative hyperplasia (NRH) upon liver biopsy. Six patients were administered intravenous immunoglobulin-G (0.4-0.6 g/kg bodyweight) once every 3-4 weeks as basic therapy. Often, CVID complicated with NCPH has: (1) The manifestations of portal hypertension as the primary symptom. (2) Autoimmune-related manifestations. Imaging can provide important diagnostic clues. The etiology may be related to hepatic NRH and splenomegaly due to recurrent infections.

Identification and molecular characterization of tea-oil camellia-associated totivirus 1.

A novel dsRNA virus was identified by high-throughput sequencing from tea oil trees in China. Its complete genome of 4714 bp contains two open reading frames (ORFs). ORF1 encodes a putative coat protein (CP) of 702 amino acids (aa), and ORF2 codes for an RNA-dependent RNA polymerase (RdRp) of 855 aa. The virus shares the highest aa sequence identity of 45.21% in RdRp with taro-associated totivirus L (MN_119621), a member of the genus Totivirus in the family Totiviridae. Phylogenetic analysis of the aa sequences of the RdRp places the new virus in a group with other totiviruses, suggesting that this virus, which is provisionally named "tea-oil camellia-associated totivirus 1", should be considered a member of the genus Totivirus.

Management and maintenance of the airway in cervical necrotising fasciitis: a retrospective analysis of 15 cases.

Cervical necrotising fasciitis is a progressive deep infection of the neck associated with high mortality, and skillful management of the airway is critical for operations under general anaesthesia. Tracheostomy under local anaesthesia has been considered the gold standard of airway management in patients with deep neck infections, but it may be difficult or impossible in advanced cases. We report here our experience over 6 years (January 2008 and December 2013) during which a total of 15 patients was diagnosed with cervical necrotising fasciitis. Of 6 patients, admitted between January 2008 and March 2010, 5 had routine tracheostomy under local anaesthesia, 1 had direct laryngoscopy intubation, and 9 who were admitted between Spring 2010 and December 2013 were treated with nasotracheal intubation. Postoperatively all patients were given moderate sedation and analgesia. Nasotracheal intubation was continued until the infection had been controlled. During intubation patency of the endotracheal tube was maintained by humidification with a continuous pump of 0.45% sodium chloride and suction. All 15 patients (10 men and 5 women, mean age 62 years, range 36-93) required an emergency drainage procedure under general anaesthesia. Fourteen of the 15 had evidence of compromise of the airway, but emergency intervention was not required. Since Spring 2010, 9 consecutive patients had required nasotracheal intubation, including 7 video laryngoscopies and 2 fibreoptic bronchoscopies. No other interventions were required. Patients were intubated postoperatively from 3 to 14 days, and there were no problems with the airway. Advanced techniques for control of the airway have a high rate of success in patients with necrotising fasciitis and could be an appropriate alternative to a traditional airway. Postoperative sedation and analgesia should be considered as routine management of pain and anxiety.

Antioxidant micronutrients improve intrinsic and UV-induced apoptosis of human lymphocytes particularly in elderly people.

OBJECTIVE: Aging and oxidative stress may lead to enhanced cellular damage and programmed cell death. To study the association of intrinsic apoptosis with age and the effect of antioxidant supplementation on intrinsic and UV-induced apoptosis in children, young and elderly people. METHODS: The study was a 2 months, double-blind, randomized trial. Three age groups were studied: children, young adults and elderly people. A total of 274 healthy subjects were allocated to a group supplemented with moderate amounts of retinol, ß-carotene, α-tocopherol, ascorbic acid and selenium or placebo. Plasma oxidative stress parameters were detected and apoptosis of lymphocytes was evaluated with TUNEL staining. RESULTS: At baseline, percentages of intrinsic apoptosis were 13.8% and 11.1% in elderly and young people, respectively, both significantly higher than children (6.3%). A decrease of 1.7% and 2.3% in intrinsic apoptosis of lymphocytes was found in the supplemented groups of young and elderly people compared with their control groups (all p values <0.001), but no significant decrease in children. Moreover, percentages UV-induced apoptosis significantly decreased by 1.4%, 1.9% and 3.1% in children, young and elderly people, respectively, compared with control groups after the trial. There were considerable increments in concentrations of plasma ß-carotene, retinol, tocopherol, ascorbic acid and selenium in all three treated groups after the supplementation. CONCLUSIONS: Young and elderly people have a higher intrinsic apoptosis than children, which was improved by antioxidant supplementation. UV-induced damage was attenuated by the supplementation in all three age groups.

Increased levels of peroxisome proliferator-activated receptor gamma, coactivator 1 alpha (PGC-1alpha) improve lipid utilisation, insulin signalling and glucose transport in skeletal muscle of lean and insulin-resistant obese Zucker rats.

AIMS/HYPOTHESIS: Reductions in peroxisome proliferator-activated receptor gamma, coactivator 1 alpha (PGC-1alpha) levels have been associated with the skeletal muscle insulin resistance. However, in vivo, the therapeutic potential of PGC-1alpha has met with failure, as supra-physiological overexpression of PGC-1alpha induced insulin resistance, due to fatty acid translocase (FAT)-mediated lipid accumulation. Based on physiological and metabolic considerations, we hypothesised that a modest increase in PGC-1alpha levels would limit FAT upregulation and improve lipid metabolism and insulin sensitivity, although these effects may differ in lean and insulin-resistant muscle. METHODS: Pgc-1alpha was transfected into lean and obese Zucker rat muscles. Two weeks later we examined mitochondrial biogenesis, intramuscular lipids (triacylglycerol, diacylglycerol, ceramide), GLUT4 and FAT levels, insulin-stimulated glucose transport and signalling protein phosphorylation (thymoma viral proto-oncogene 2 [Akt2], Akt substrate of 160 kDa [AS160]), and fatty acid oxidation in subsarcolemmal and intermyofibrillar mitochondria. RESULTS: Electrotransfection yielded physiologically relevant increases in Pgc-1alpha (also known as Ppargc1a) mRNA and protein ( approximately 25%) in lean and obese muscle. This induced mitochondrial biogenesis, and increased FAT and GLUT4 levels, insulin-stimulated glucose transport, and Akt2 and AS160 phosphorylation in lean and obese animals, while bioactive intramuscular lipids were only reduced in obese muscle. Concurrently, PGC-1alpha increased palmitate oxidation in subsarcolemmal, but not in intermyofibrillar mitochondria, in both groups. In obese compared with lean animals, the PGC-1alpha-induced improvement in insulin-stimulated glucose transport was smaller, but intramuscular lipid reduction was greater. CONCLUSIONS/INTERPRETATIONS: Increases in PGC-1alpha levels, similar to those that can be induced by physiological stimuli, altered intramuscular lipids and improved fatty acid oxidation, insulin signalling and insulin-stimulated glucose transport, albeit to different extents in lean and insulin-resistant muscle. These positive effects are probably attributable to limiting the PGC-1alpha-induced increase in FAT, thereby preventing bioactive lipid accumulation as has occurred in transgenic PGC-1alpha animals.

Coordinately regulated expression of FAT/CD36 and FACS1 in rat skeletal muscle.

Protein-mediated fatty acid uptake and intracellular fatty acid activation are key steps in fatty acid metabolism in muscle. We have examined (a) the abundance of fatty acid translocase (FAT/CD36) mRNA (a fatty acid transporter) and long-chain acyl CoA synthetase (FACS1) mRNA in metabolically heterogeneous muscles (soleus (SOL), red (RG) and white gastrocnemius (WG)), and (b) whether FAT/CD36 and FACS1 mRNAs were coordinately up-regulated in red (RTA) and white tibialis muscles (WTA) that had been chronically stimulated for varying periods of time (0.25, 1, 6 and 24 h/day) for 7 days. FAT/CD36 mRNA and FACS1 mRNA abundance were scaled with (a) the oxidative capacity of muscle (SOL > RG > WG) (p < 0.05), (b) the rates of fatty acid oxidation in red and white muscles, and (c) fatty acid uptake by sarcolemmal vesicles, derived from red and white muscles. In chronically stimulated muscles (RTA and WTA), FAT/CD36 mRNA and FACS1 mRNA were up-regulated in relation to the quantity of muscle contractile activity (p < 0.05). FAT/CD36 mRNA and FACS1 mRNA up-regulation was highly correlated (r = 0.98). The coordinated expression of FAT/CD36 and FACS is likely a functional adaptive response to facilitate a greater rate of fatty acid activation in response to a greater rate of fatty acid transport, either among different types of muscles or in muscles in which capacity for fatty acid metabolism has been enhanced.

Epinephrine translocates GLUT-4 but inhibits insulin-stimulated glucose transport in rat muscle.

We examined the effects of epinephrine (25, 50, and 150 nM) on 1) basal and insulin-stimulated 3-O-methylglucose (3-MG) transport in perfused rat muscles and 2) GLUT-4 in skeletal muscle plasma membranes. Insulin increased glucose transport 330-600% in three types of skeletal muscle [white (WG) and (RG) gastrocnemius and soleus (SOL)]. Glucose transport was also increased by epinephrine (22-48%) in these muscles (P < 0.05). In contrast, the insulin-stimulated 3-MG transport was reduced by epinephrine in all three types of muscles; maximal reductions were observed at 25 nM epinephrine in WG (-25%) and RG (-32.5%). A dose-dependent decrease occurred in SOL (-27% at 25 nM; -55% at 150 nM, P < 0.05). Insulin (20 mU/ml) and epinephrine (150 nM) each translocated GLUT-4 to the plasma membrane, and no differences in translocation were observed between insulin and epinephrine (P > 0.05). In addition, epinephrine did not inhibit insulin-stimulated GLUT-4 translocation, and the combined epinephrine and insulin effects on GLUT-4 translocation were not additive. The increase in surface GLUT-4 was associated with increases in muscle cAMP concentrations, but only when epinephrine alone was present. No relationship was evident between muscle cAMP concentrations and surface GLUT-4 in the combined epinephrine and insulin-stimulated muscles. These studies indicate that epinephrine can translocate GLUT-4 while at the same time increasing glucose transport when insulin is absent, or can inhibit glucose transport when insulin is present.

Increments in skeletal muscle GLUT-1 and GLUT-4 after endurance training in humans.

We investigated the time course of training-induced changes in the expression of GLUT-1 and GLUT-4 in human skeletal muscle. Seven healthy males trained for 2 h/day (approximately 60% pretraining VO2peak) for 31 days (31D). Muscle biopsies were obtained before training (PRE) and after 5 (5D) and 31 days (31D) of training. Training resulted in progressive increases in muscle GLUT-4 with increasing training duration (PRE<5D<31D; P<0.01). Muscle GLUT-1 content was also increased (P<0.05) after training; however, the increase was not observed until 31D (131%). Increases in muscle hexokinase (HK) activity were complete by 5D (P<0.01). Muscle malate dehydrogenase activity was not elevated after 5D of training but was increased (+35%; P<0.01) at 31D. Results from this study show that increases in both GLUT-4 and HK represent early training-induced adaptations to prolonged exercise training. As training progresses, further increases in GLUT-4, but not HK, occur in conjunction with an increase in muscle mitochondrial potential and GLUT-1.

[Effect of cyclic nucleotides (cAMP) on experimental HSV-I keratitis].

The authors report an experimental study on the effect of cAMP on HSV-I keratitis. Rabbit corneas developed typical dendritic keratitis on the 3rd day after inoculation of HSV-I. On the 9th day, the cAMP level in plasma significantly decreased from 174.9 +/- 20.2 to 52.1 +/- 18.2 pmol/ml, while the cGMP level increased. The cAMP level in aqueous humour also decreased, with an increase in cGMP level increased. The cAMP level in aqueous humour also decreased, with an increase in cGMP level. cAMP was used subconjunctivally for treatment of HSV-I keratitis. 48 hours after injection, the cAMP level in aqueous humour markedly rose from 11.1 +/- 2.0 to 35.6 +/- 12.9 pmol/ml and 6 days later to 65.0 +/- 30.9 pmol/ml, with concurrent decrease in cGMP level, and the ratio of cAMP/cGMP increased from 1.6 +/- 0.6 to 15.3 +/- 4.6. In the group of treatment by acyclovir, the level of cAMP in aqueous humour increased by 9.5 times after 3 days of treatment, with no remarkable effect on the cGMP level. These results showed that cAMP was equally effective as ACV in treatment of experimental HSV 1 keratitis. The antiviral action of cAMP was discussed and the pharmacological effect of cAMP approached.