Identification of the GAPDH gene family in Citrullus lanatus and functional characteristics of ClGAPC2 in Arabidopsis thaliana.

Members of the GAPDH family play important roles in plant growth and development, as well as in stress responses. Our aim was to identify stress resistance genes through systematic analysis of the GAPDH family in watermelon. This could not only provide genetic resources for stress resistance breeding, but also form a basis for the study of plant stress resistance mechanisms. Eight GAPDHs representing four types of plant GAPDH in watermelon were identified (ClGAPA/B, ClGAPC1-3, ClGAPCp1-2 and ClGAPN). A comprehensive analysis of physicochemical properties, chromosome distribution, evolutionary relationships, exon-intron structure and conserved motifs of watermelon GAPDHs was performed using bioinformatics. Expression characteristics were assessed by RT-qPCR. Based on RT-qPCR results, ClGAPC2 was screened as a candidate for subcellular localization analysis and functional verification in Arabidopsis thaliana. Eight GAPDHs were classified into four subfamilies. GAPDHs in each subgroup were generally conserved and shared similarities in structure and conserved motifs. ClGAPDHs had notable tissue specificity and different expression patterns in response to H2 O2 , chilling, salt, osmotic stress, heat, salicylic acid, gibberellin, brassinosterol, ethylene and abscisic acid treatments. Three ClGAPC genes, especially ClGAPC2, were markedly induced by several treatments. ClGAPC2 was located in the nucleus and cytoplasm of tabacum epidermal cells. The ClGAPC2 transgenic Arabidopsis showed enhanced tolerance to salinity at the germination stage. We suggest that ClGAPC2 plays important roles in the adaptation of watermelon to salinity. Our findings provided candidate genes for further improving the salt tolerance of watermelon.

[Clinical features and treatment outcome of patients with non-Hodgkin lymphoma-associated hemophagocytic lymphohistiocytosis].

Objective: To investigate the clinical features and effect of prognostic factors in patients with different pathological types of non-Hodgkin lymphoma-associated hemophagocytic lymphohistiocytosis. Methods: We collected and analyzed the clinical data of 89 patients with non-Hodgkin lymphoma-associated hemophagocytic lymphohistiocytosis who were treated at Huadong Hospital from March 2013 to May 2020. The data were analyzed via log-rank and Cox multivariate analyses. Results: The median overall survival time of the 89 cases was 10.2 months. Patients with B-cell lymphoma-associated hemophagocytic lymphohistiocytosis did not reach the median overall survival time. The median overall survival times of T-cell lymphoma-associated hemophagocytic lymphohistiocytosis and NK-cell lymphoma-associated hemophagocytic lymphohistiocytosis were 10.2 and 3.0 months, respectively. The pathological type of non-Hodgkin lymphoma (OS: P=0041, PFS: P=0.015) , ECOG score ≥ 3 (OS: P=0.031, PFS: P=0.030) , hematopoietic stem cell transplantation (OS: P=0.005, PFS: P=0.040) , lymphadenopathy (OS: P=0.007, PFS: P=0.012) , and splenomegaly (OS: P=0.276, PFS: P=0.324) were related to the overall survival and progression-free survival of patients with non-Hodgkin lymphoma-associated hemophagocytic lymphohistiocytosis. Splenectomy could improve the prognosis of patients with lymphoma-associated hemophagocytic lymphohistiocytosis, especially T-cell lymphoma-associated hemophagocytic lymphohistiocytosis. Conclusion: The clinical characteristics of patients with different pathological types of non-Hodgkin lymphoma-associated hemophagocytic lymphohistiocytosis were similar but were different in the overall survival rate and the effect of prognostic factors. We suggested that patients with non-Hodgkin lymphoma-associated hemophagocytic lymphohistiocytosis should receive more than combined chemotherapy. To improve the prognosis and survival rate of patients, those with B-cell lymphoma-associated hemophagocytic lymphohistiocytosis and NK-cell lymphoma-associated hemophagocytic lymphohistiocytosis promptly require hematopoietic stem cell transplantation. Moreover, patients with T-cell lymphoma-associated hemophagocytic lymphohistiocytosis should consider splenectomy.

Expression of long non-coding RNA linc-ITGB1 in breast cancer and its influence on prognosis and survival.

OBJECTIVE: Long noncoding RNA linc-ITGB1 (linc-ITGB1) was reported to serve as a tumor promoter in breast cancer (BC). However, the clinical significance of linc-ITGB1 has not been reported. The present study aimed to determine the relationship between linc-ITGB1 expression and clinicopathological features and survival. PATIENTS AND METHODS: qRT-PCR was used to quantify the expression levels of linc-ITGB1 in BC and adjacent non-cancerous breast tissues. The X2 test was performed to determine the associations between linc-ITGB expression and the clinicopathological characters. The overall survival time (OS) and disease-free survival (DFS) were collected by follow-up and analyzed by Kaplan-Meier analysis. Multivariate Cox regression analysis was used to identify the independent risk factors for BC. RESULTS: The results showed that linc-ITGB1 levels were lower in tumor tissues of BC patients in comparison to adjacent non-cancerous breast tissues (p < 0.001). Linc-ITGB1 expression was significantly associated with lymph node metastasis, pathological differentiation and TNM stage (all p < 0.05). Furthermore, Kaplan-Meier analysis demonstrated that high-linc-ITGB1 expression level was associated with poorer OS (p = 0.006) and DFS (p = 0.003). Cox proportional hazards risk analysis demonstrated that linc-ITGB1 was an independent predictor for both OS (p = 0.004) and DFS (p = 0.002) in BC. CONCLUSIONS: These results indicated, for the first time, that linc-ITGB1 be a potential biomarker in the prognosis of BC.

Down-regulation of AQP4 Inhibits Proliferation, Migration and Invasion of Human Breast Cancer Cells.

Aquaporins (AQPs), proteinaceous water channels, have been proposed as mediators of tumour development and progression. However, the role of aquaporin 4 (AQP4), a member of the AQP family, in breast cancer has not been distinctly evaluated. The aim of the present study was to examine the effect of AQP4 down-regulation on proliferation, migration and invasion in human breast cancer. To determine this effect, siRNA interference was used to knock down its expression in T47D and MCF-7 cell lines. Down-regulation of AQP4 resulted in increased expression of E-cadherin along with an inhibitory effect on the proliferation, migration and invasion in breast cancer cells. In addition, AQP4 regulation of cell proliferation could be related with the ERK/Ecadherin pathway. In conclusion, the present data have suggested that down-regulation of AQP4 inhibits breast cancer cell proliferation, migration and invasion.

[Clinicopathological characteristics and prognosis of different molecular types of breast cancer].

OBJECTIVE: To investigate the clinicopathological characteristics of 4 subtypes of breast cancer, Luminal A, Luminal B, human epidermal growth factor receptor 2(HER2), triple-negative and their associated prognostic factors. METHODS: The clinical characteristics and prognosis of 102 patients with breast cancer who treated in Wuhan University Renmin Hospital from January 2011 to November 2012 were retrospectively analyzed. RESULTS: The proportion of each subtype was Luminal A 27.5%, Luminal B 35.3%, HER2 14.7%, and triple-negative 22.5%. The age range was from 22 to 75 years old, with median age of 48.0 years old. All patients underwent surgery and 98 cases were also treated with chemotherapy/endocrine+ Herceptin therapy (96.1%)after surgery. The tumor size, histological grade, and lymph node metastasis had significant difference in different subtypes of breast cancer (P<0.05). Further analysis showed the proportion of HER2 tumor≤2 cm (4, 26.7%)had a tendency of smaller than that in luminal A(20, 71.4%) or triple negative(16, 69.6%) (P=0.009>0.008, P=0.019>0.008). Histological Ⅲ grade proportion in patients with HER2 subtype (8, 53.3%) was in greater tendency than that with Luminal A subtype(23, 82.1%) (P=0.039>0.008). The proportion of patients with no metastatic lymph nodes in triple negative subtype(7, 30.4%) had a tendency of smaller than that in HER2 subtype(7, 46.7%) or Luminal B subtype(17, 47.2%)(P=0.019>0.008, P=0.016>0.008). There were no significant differences in age of onset, menstruation status, operation, family cancer history, and the risk of recurrence and metastasis in different subtypes (P>0.05). In Luminal A subtype, three-years overall survival rate was 100%, and one-, two-, three-year event-free survival rate were all 92.9%. In Luminal B subtype, three-year overall survival rate was 100%, event-free survival rate at one-, two- and three-year were all 97.2%. In HER2 subtype, three-year overall survival rate was 100%, one-, two- and three-year event-free survival rate were 100%, 100%, and 93.3%, respectively. The 1, 2, 3 years overall survival rate of triple-negative subtype were 100%, 95.7%, and 95.7%, respectively. One-, two- and three-year event-free survival rate of triple-negative subtype were 95.7%, 91.3%, and 90.0% respectively. Whether triple negative subtype, recurrence risk, age of onset, menstrual status, tumor size, histological grade, lymph node metastasis, surgery, breast cancer or other family history of cancer were not significantly related with three-year event-free survival rate (P>0.05). CONCLUSIONS: Luminal subtype had the largest proportion and better prognosis. The proportion of tumor >2 cm and histological grade Ⅲ in patients with HER2 subtype was larger, and three-year event-free survival rate of this patients was lower. Triple-negative subtype had a greater tendency of lymph node metastasis and the lowest three-year event-free survival. Tumor size, histological grade and lymph node metastasis may be the important factors for the prognosis of breast cancer.

[Radiation therapy in thyroid cancer]. / Place de la radiothérapie externe dans les cancers de la thyroïde.

Anaplastic thyroid cancers represent 1-2% of all thyroid tumours and are of very poor prognosis even with multimodality treatment including external beam radiation therapy. Conversely, differentiated thyroid carcinomas (at least 80% of thyroid cancers) hamper good prognosis with surgery with or without radioiodine and there is hardly any room for external beam radiation therapy. Insular and medullar carcinomas have intermediary prognosis and are rarely irradiated. We aimed to update recommendations for external beam irradiation in these different clinical situations and put in light the benefits of new irradiations techniques. A search of the French and English literature was performed using the following keywords: thyroid carcinoma, anaplastic, chemoradiation, radiation therapy, surgery, histology and prognostic. Non-mutilating surgery (often limited to debulking) followed by systematic external beam radiation therapy is the standard of care in anaplastic thyroid cancers (hyperfractionated-accelerated radiation therapy with low-dose weekly doxorubicin with or without cisplatin if possible). Given anaplastic thyroid cancers' median survival of 10 months or less, neoadjuvant and adjuvant chemotherapy may also be discussed. Ten-year survival rates for patients with papillary, follicular and Hürthle-cell carcinomas are 93%, 85%, and 76%, respectively. Massive primary incompletely resected iodine-negative disease indicates external beam radiation therapy. Older age (45 or 60-year-old), poor-prognosis histological variants (including tall cell cancers) and insular cancers are increasingly reported as criteria for external beam radiation therapy. Massive extracapsular incompletely resected nodal medullary disease suggests external beam radiation therapy. Radiation therapy morbidity has been an important limitation. However, intensity modulated radiation therapy (IMRT) offers clear dosimetric advantages on tumour coverage and organ sparing, reducing late toxicities to less than 5%. The role of radiation therapy is evolving for anaplastic thyroid cancers using multimodal strategies and new chemotherapy molecules, and for differentiated cancers using minor criteria, such as histological variants, with IMRT becoming a standard of care.

Indications of external beam radiation therapy in non-anaplastic thyroid cancer and impact of innovative radiation techniques.

BACKGROUND: The mainstay of treatment for differentiated thyroid carcinomas is surgery. There is hardly any room for radiation therapy in differentiated thyroid carcinomas. We aimed to update recommendations for RT in the context of histological variants, increased use of radioiodine and new irradiations techniques. MATERIALS AND METHODS: A search of the French and English literature was performed using thyroid carcinoma, radiation therapy, surgery, variants and radioiodine. RESULTS: Papillary, follicular, Hürthle and medullary carcinomas represent about 80%, 11%, 3% and 4% of all thyroid carcinomas, respectively. Ten-year survival rates for patients with papillary, follicular and Hürthle cell carcinomas are 93%, 85%, and 76%, respectively. The occurrence of criteria such as older age (45 or 60 years-old), massive primary disease, extensive extracapsular spread and macroscopic iodine-negative components inconsistently indicate external beam irradiation (EBRT). The impact of EBRT on poorer-prognosis histological variants is an emerging issue. Noteworthy, the incidence of laryngeal and wound healing complications has been an important limitation to EBRT. However, intensity modulated radiation therapy (IMRT) offers clear dosimetric advantages on tumor coverage and organ sparing such as the larynx, thus reducing late toxicities to less than 5%. Iodine contrast agents should be avoided during 4-6 weeks before radioiodine. PET CT is increasingly used in iodine-negative tumors. CONCLUSION: There are elective indications for EBRT and IMRT has the potential to improve local control.

Chemoradiation in anaplastic thyroid carcinomas.

BACKGROUND: ATC represents 1-2% of all thyroid carcinomas. Median survival is poor (3-10 months). Our goal is to update recommendations for RT in the context of new irradiation techniques. MATERIALS AND METHODS: A search of the French and English literature was performed with terms: thyroid carcinoma, anaplastic, chemoradiation, radiation therapy and surgery. Level-based evidence remains limited in the absence of prospective studies and the small size of retrospective series of this rare tumor. RESULTS: Surgery when possible should be as complete as possible but without mutilation given the 8-month median survival of ATC. It should be followed by systematic chemoradiation in ATC. Initiation of treatment is an emergency given fast tumor doubling time. The most promising results of chemoradiation to date have been shown in series of radiation therapy (+/- acceleration) combined with doxorubicin +/- taxanes or cisplatin. Adjuvant chemotherapy (doxorubicin, cisplatine and/or taxane-based) may also be recommended given the metastatic potential of ATC and warrants further investigations. Data on neoadjuvant chemotherapy are missing. Intensity modulated radiation therapy offers clear dosimetric advantages and has the potential to improve tumor and nodal (posterior neck, mediastinum) coverage, i.e., locoregional control while optimally sparing the spinal cord, larynx, parotids, trachea and esophagus. PET-CT and MRI may be used for RT planning. CONCLUSION: Chemoradiation with debulking surgery whenever possible is the mainstay of treatment of anaplastic thyroid carcinomas (ATC). EBRT using IMRT has the potential to improve local control. Taxane-doxorubicin concomitant chemoradiotherapy is worth further investigation.

Comparison of the therapeutic effects of leflunomide and mycophenolate mofetil in the treatment of immunoglobulin A nephropathy manifesting with nephrotic syndrome.

OBJECTIVE: To evaluate the therapeutic effects and safety of a combination therapy of leflunomide (LEF) and prednisone for the treatment of immunoglobulin A (IgA) nephropathy manifesting with nephrotic syndrome. METHODS: 40 patients with IgA nephropathy manifesting with nephrotic syndrome were randomly divided into two groups. The treatment group was administered with a combination therapy of prednisone and LEF, and the control group with a combination therapy of prednisone and MMF. For the following comparison 24-h urinary protein excretion and the serum levels of albumin, cholesterol, and creatinine before and after the therapy were assessed. RESULTS: In the treatment group, the medication was markedly effective in 5 cases and effective in 7 cases; the total efficacy rate was 60.0%. In the control group, the treatment was markedly effective in 5 cases and effective in 8 cases; the total efficacy rate was 65.0%. The IgA levels in both groups decreased after therapy. There were no significant differences between the results for the two groups (p > 0.05). The treatments were well tolerated in both groups. CONCLUSION: LEF is a safe and effective drug for the treatment of IgA nephropathy manifesting with nephrotic syndrome.

Inhibitory effects of low-dose polymyxin B on hemorrhage-induced endotoxin/bacterial translocation and cytokine formation in rats.

The current experiments were performed to determine the effects of a subtherapeutic dose of polymyxin B sulfate on gut origin endotoxemia/bacterial translocation, and tumor necrosis factor (TNF) and interleukin-1 (IL-1) release following hemorrhagic shock (30 mm Hg, 90 min) in rats. The results showed that significant portal and systemic endotoxemia took place in the control group (portal, 0.269 to 0.845 endotoxin units (EU)/mL; systemic, 0.164 to 0.655 EU/mL), but not in the treatment group (except 0.5 hour in portal blood: 0.207 +/- 0.094 EU/mL). Concomitantly, the incidence of bacterial translocation to the mesenteric lymph nodes and viscera were reduced significantly at 0.5, 2, 6, and 24 hours postresuscitation in animals receiving polymyxin B (p < 0.05 to 0.01), whereas there were no differences with respect to number of translocating bacteria between the two groups (p > 0.05). Marked elevation of plasma TNF levels and IL-1 activities of peritoneal macrophages were also found in untreated controls at 0.5 to 2 hours (p < 0.05) and 6 to 24 hours (p < 0.05 to 0.01), respectively, but prevented by administration of low-dose polymyxin B. The 48-hour survival rate was improved from 41.7% in the control group to 75.0% in the treatment ones (p > 0.05). These data suggest that pretreatment with a subtherapeutic dose of polymyxin B is effective to inhibit hemorrhage-induced endotoxin/bacterial translocation from the gut and excessive TNF and IL-1 production.

[Changes in expression of interleukin-2 and interleukin-2 receptor in lymphocytes in patients with burns].

IL-2 mainly combines with the IL-2R on the membrane of the lymphocyte, both to promote activation and proliferation. The expression of IL-2 and IL-2R on the lymphocyte was found to be significantly depressed after burn injury. IL-2 was decreased by 40%-60%, and the expression of IL-2R was decreased by 25%-55%, especially on the 7th day after burn. The results demonstrated that burn injury not only influenced the production of IL-2 from T helper cell, but also depressed the expression of IL-2R.

Electron-capture GLC determination of pseudoephedrine in serum.

A GLC assay was developed for pseudoephedrine in serum using 3-methylamino-1-phenyl-1-propanol as an internal standard. After extraction from serum with benzene under alkaline conditions, pseudoephedrine was derivatized with pentafluorobenzyl bromide and quantitated by electron-capture GLC. The method has a detection limit of approximately 0.02 microgram/ml of serum using 1-ml samples.

Rapid determination of theophylline in serum by electron-capture GLC.

A rapid and sensitive GLC procedure was developed for the determination of theophylline in serum. After extraction from serum with ethyl acetate, theophylline and the internal standard were derivatized with pentafluorobenzyl bromide under alkaline conditions. The derivatives were quantitated by electron-capture detection. The method has a sensitivity of 0.1 microgram/ml with a 0.1-ml serum sample.

Rapid, sensitive GLC determination of pentobarbital and other barbiturates in serum using nitrogen-specific detector.

A GLC method for the analysis of pentobarbital in serum was developed. After extraction from serum, a methyl derivative was prepared and quantitated by nitrogen-specific detection. The method has a senstivity of 0.08 microgram/ml for pentobarbital with only 0.1 ml of serum. Secobarbital was the internal standard. Derivatives of other barbiturates also were prepared.

Rapid GLC determination of chlordiazepoxide and metabolite in serum using on-column methylation.

A rapid GLC method was developed for the assay of chlordiazepoxide in serum. After chlordiazepoxide was extracted with ether, it was methylated with trimethylanilinium hydroxide in the injection port and detected by electron capture. The assay is simple and sensitive and can be automated for large-scale clinical analysis.

Fluorescence--TLC densitometric determination of diazepam and other 1,4-benzodiazepines in serum.

A sensitive fluorescence--TLC densitometric procedure was developed for the specific determination of diazepam (I) and its two metabolites, desmethyldiazepam (II) and oxazepam (III), in serum. After extraction from serum with benzene, the compounds were separated by TLC and converted with a sulfuric acid spray to greenish-yellow fluorescence spots with Rf values of 0.72, 0.43, and 0.17, respectively. Quantitation of the TLC plate was accomplished by scanning with a densitometer at 380 nm. The sensitivities of the assay were 18 (I), 6 (II), and 5 (III) ng/ml of serum. This procedure was also was applicable to other 1,4-benzodiazepines in biological fluids.

Quantitative determination of chlordiazepoxide and its metabolites in serum by fluorescence TLC--densitometry.

A sensitive fluorescence TLC--densitometric procedure was developed for the specific determination of chlordiazepoxide (I) and its two metabolites, demoxepam (II) and desmethylchlordiazepoxide (III), in serum. After extraction from serum with ether, I, II, and II were separated by TLC and converted with a sulfuric acid spray to pale blue (Rf 0.63), green (Rf 0.54), and blue (Rf 0.45) fluorescence spots, respectively. Quantitation was accomplished by scanning the plate with a densitometer at 390 (I), 430 (II), and 390 (III) nm. The sensitivities were 0.05 (I), 0.01 (II), and 0.01 (III) microgram/ml of serum. The procedure was successfully applied to measurement of I-III in human serum after oral administration of 20 mg of chlordiazepoxide hydrochloride.

Determination of pentobarbital in serum by electron-capture GLC.

A GLC method was developed for pentobarbital in serum. After extraction from serum, a pentafluorobenzyl derivative was prepared and quantitated by electron-capture detection. The method has a sensitivity of 0.1 microng/ml of serum, and the amount detectable is less than 0.2 ng/injection. Hexethal was used as the internal standard. Derivatives of other barbiturates were also made. NMR and mass spectrometric analyses confirmed the proposed structure of the 1,3-bis(pentafluorobenzyl) derivative of pentobarbital. The procedure was successfully applied to measurement of serum pentobarbital levels in humans.