Application of NIRS for hay evaluation at different degrees of sample preparation.

Objective: A study compared the performance of the NIRS (Near-Infrared Spectroscopy) calibration models developed with different degrees of hay sample preparations. Methods: Spectral data of 1-mm ground or whole hay samples were regressed against wet chemistry results of moisture, NDF (neutral detergent fiber), ADF (acid detergent fiber), CP (crude protein), and IVDMD (in vitro dry matter digestibility). A total of 227 imported alfalfa (Medicago sativa L.) and another 360 timothy (Phleum pratense L.) hay samples were used to develop the calibration models. The models developed with ground hay samples were more robust and accurate than whole hay based on cross-validation's R2 (coefficient of determination), standard error, and RPD (ratio percentage deviation). Results: The R2 of cross-validation ranged from 0.61 (moisture of alfalfa) to 0.95 (CP prediction of timothy). Although R2 of calibration models was mainly greater than 0.90, the R2 of cross-validations remained marginal. Conclusion: Estimation of nutrient concentrations in imported hay can be achieved by calibrated NIRS. The NIRS calibration models must be improved by including more imported hay samples from different years and origins. Although the analysis accuracy of NIRS was substantially higher when calibration models were developed with ground samples, less sample preparation will be more advantageous for achieving rapid delivery of hay sample analysis results. Therefore, further research warrants investigating the level of sample preparation inputs compromising analysis accuracy by NIRS.

Diagnostic Efficacy of 18F-FDG PET/CT in Detecting Bone Marrow Infiltration in Patients with Newly Diagnosed Diffuse Large B-Cell Lymphoma.

Objective: Diffuse large B-cell lymphoma (DLBCL) is often associated with bone marrow infiltration, and 2-deoxy-2-(18F) fluorodeoxyglucose positron emission tomography/computed tomography ( 18F-FDG PET/CT) has potential diagnostic significance for bone marrow infiltration in DLBCL. Methods: A total of 102 patients diagnosed with DLBCL between September 2019 and August 2022 were included. Bone marrow biopsy and 18F-FDG PET/CT examinations were performed at the time of initial diagnosis. Kappa tests were used to evaluate the agreement of 18F-FDG PET/CT with the gold standard, and the imaging features of DLBCL bone marrow infiltration on PET/CT were described. Results: The total detection rate of bone marrow infiltration was not significantly different between PET/CT and primary bone marrow biopsy ( P = 0.302) or between the two bone marrow biopsies ( P = 0.826). The sensitivity, specificity, and Youden index of PET/CT for the diagnosis of DLBCL bone marrow infiltration were 0.923 (95% CI, 0.759-0.979), 0.934 (95% CI, 0.855-0.972), and 0.857, respectively. Conclusion: 18F-FDG PET/CT has a comparable efficiency in the diagnosis of DLBCL bone marrow infiltration. PET/CT-guided bone marrow biopsy can reduce the misdiagnosis of DLBCL bone marrow infiltration.

Single-center phase 2 study of PD-1 inhibitor combined with DNA hypomethylation agent + CAG regimen in patients with relapsed/refractory acute myeloid leukemia.

Anti-PD-1 monotherapy had limited clinical efficacy in relapsed/refractory (r/r) AML patients with higher PD-1 and PD-L1 expression. Hence, we investigated the efficacy and safety of PD-1 inhibitor with DNA hypomethylating agent (HMA) + CAG regimen in patients who had failed prior AML therapy. In this phase 2, single-arm study, r/r AML patients received azacitidine or decitabine plus CAG regimen with tislelizumab. Primary endpoints were efficacy (objective response rate [ORR]) and safety. Secondary endpoints included overall survival (OS), event-free survival (EFS) and duration of response (DOR). Statistical analyses were performed using Stata 14.0 and SPSS 20.0 software where P < 0.05 denoted significance. Twenty-seven patients were enrolled patients and completed 1 cycle, and 14 (51.9%) and 4 (14.8%) patients completed 2 and 3 cycles, respectively. ORR was 63% (14: complete remission [CR]/CR with incomplete hematologic recovery [CRi], 3: partial remission (PR), 10: no response [NR]). Median OS (mOS) and EFS were 9.7 and 9.2 months, respectively. With a median follow-up of 8.2 months (1.1-26.9), the mOS was not reached in responders (CR/CRi/PR) while it was 2.4 months (0.0-5.4) in nonresponders (P = 0.002). Grade 2-3 immune-related adverse events (irAEs) were observed in 4 (14.8%) patients and 3 nonresponders died of lung infection after treatment. Tislelizumab + HMA + CAG regimen showed improved outcomes in r/r AML patients with lower pretherapy leukemia burden. irAEs were mild and low-grade and higher pretherapy bone marrow CD4+ CD127+ PD-1+ T cells might serve as a predictor of treatment response.ClinicalTrials.gov identifier NCT04541277.

Effects of sodium diacetate and microbial inoculants on fermentation of forage rye.

Rye (Secale cereale L.) is a valuable annual forage crop in Korea but there is limited information about the impact of chemical and biological additives on fermentation characteristics of the crop. This experiment was conducted to investigate fermentation dynamics of wilted forage rye treated with the following six additives; control (no additive), sodium diacetate applied at 3 g/kg wilted forage weight (SDA3), 6 g/kg wilted forage weight (SDA6), inoculations (106 CFU/g wilted forage) of Lactobacillus plantarum (LP), L. buchneri (LB), or LP+LB. The ensiled rye sampled at 1, 2, 3, 5, 10, 20, 30, and 45 days indicated that the acidification occurred fast within five days of storage than the rest of the storage period. The microbial inoculants decline the pH of ensiled forage, more rapidly than the control or SDA treated, which accompanied by the decrease of water-soluble carbohydrates and increase of lactic acid. Compared with the control silage, all treatments suppressed ammonia-nitrogen formation below to 35 g/kg DM throughout the sampling period. Suppression of total microbial counting occurred in SDA6, LP, and LP + LB. The lactic acid production rates were generally higher in microbial inoculation treatments. Acetic acid concentration was lowest in the LP-treated silage and highest in the SDA- and LB-treated silages. The in vitro dry matter (DM) digestibility and total digestible nutrients were the highest in the silage treated with SDA (6 g/kg) at day 45 of ensiling. Based on lower ammonia-nitrogen concentrations and higher feed value, ensiling forage rye treated with SDA at 6 g/kg is promising through enhanced silage quality.

[Characteristics of Phytoplankton Community Structure and Their Relationships with Environmental Factors in Autumn in Qinhe River Basin of Jincheng Region].

In October-November 2020, the phytoplankton and the aquatic environment from 62 sites in the mainstream of the Qinhe River and the largest tributary of the Qinhe River (Danhe River) in the Jincheng region were investigated to clarify the spatial pattern of phytoplankton communities and their driving factors. A total of 7 phyla and 47 species of phytoplankton were identified in the Qinhe River basin and were composed of Cryptophyta, Chlorophyta, Pyrrophyta, Chrysophyta, Bacillariophyta, Cyanophyta, and Crytophyta. Six dominant species in the Qinhe River included:Chlorella vulgaris, Cryptomonas erosa, Chroomonas acuta, Cyclotella stelligera, Chlorococcum, and Euglena viridis. Six dominant species in the Danhe River included:C. erosa, Frustulia vulgaris, E. viridis, C. vulgaris, Trachelomonas oblonga Lemm, and C. stelligera. The Shannon-Wiener diversity index (H') varied from 0.35 to 3.15, with a mean value 1.40. The Pielou evenness index (J) varied from 0.24 to 1.00, with a mean value of 0.68. H' values in the Qinhe River were higher than those in the Danhe River. J values were relatively low in the middle reaches of the Qinhe River and middle-low reaches of the Danhe River. The results in the Qinhe River through a canonical correspondence analysis (CCA) showed that the percent of forest land at a 300 m buffer was the driving factor of Chlorococcum in Chlorophyta, and nitrate, total phosphorus, and the percent of forest land at the 300 m buffer were the driving factors of E. viridis. Cyclotella stelligera was mainly influenced by the percent of urban land and water temperature, whereas C. vulgaris, C. erosa, and C. acuta were mainly influenced by the percent of farmland and residential land at the 300 m buffer. The results in the Danhe River via CCA showed that C. erosa and C. stelligera were mainly influenced by pH and sulfate, E. viridis was mainly influenced by the percent of urban land and grass land, T. oblonga Lemm was mainly influenced by chloride and the percent of forest land, F. vulgaris was mainly influenced by water temperature and the percent of farmland, and C. vulgaris was mainly influenced by ammonia and the percent of farmland.

Effects of sodium diacetate or microbial inoculants on aerobic stability of wilted rye silage.

OBJECTIVE: The primary goal was to identify the effectiveness of chemical or biological additives in delaying the deterioration of early-harvested wilted rye silage after exposure to air. METHODS: Rye harvested as a whole plant at the early heading stage was wilted for 24 h. The wilted forage was divided into treatments including sodium diacetate (SDA) at 3 (SDA3) and 6 g/kg (SDA6), Lactobacillus plantarum (LP), L. buchneri (LB), or their equal mixture (LP+LB) at 1×106 colony-forming unit/g fresh matter. RESULTS: After 60 d of conservation in 20-L silos, lactic acid was greater in LP and LP+LB silages than other treatments (102 vs 90.2 g/kg dry matter [DM]). Acetic acid was greatest in SDA6 (32.0 g/kg DM) followed by LB (26.1 g/kg DM) and was lowest in LP treatment (4.73 g/kg DM). Silage pH was lower with microbial inoculation and the lowest and highest values were observed in LP and untreated silages, respectively. After 60 d, neutral detergent fiber concentration was lowest in SDA6 silages, resulting in the greatest in vitro DM digestibility (846 g/kg DM). Aerobic stability was longest in SDA6 (176 h) followed by LB treatment (134 h). Instability after aerobiosis was greatest in LP silages (68 h), about 8 h less than untreated silages. After aerobic exposure, yeast and mold numbers were lowest in SDA6 silages, resulting in DM loss minimization. Exhaustion of acetic acid and lactic acid after aerobic exposure was lowest with SDA6 but greatest with untreated and LP silages. CONCLUSION: Treatment of early-cut wilted rye forage with SDA at 6 g/kg resulted in silages with higher feeding value and fermentation quality, and substantially delayed deterioration after aerobic exposure, potentially qualifying SDA at this load for promotion of silage quality and delaying aerobic spoilage of early-harvested (low DM) rye forage.

Evaluation of forage production, feed value, and ensilability of proso millet (Panicum miliaceum L.).

Whole-plant corn (Zea may L.) and sorghum-sudangrass hybrid [Sorghum bicolor (L.) Moench] are major summer crops that can be fed as direct-cut or silage. Proso millet is a short-season growing crop with distinct agronomic characteristics that can be productive in marginal lands. However, information is limited about the potential production, feed value, and ensilability of proso millet forage. We evaluated proso millet as a silage crop in comparison with conventional silage crops. Proso millet was sown on June 8 and harvested on September 5 at soft-dough stage. Corn and sorghum-sudangrass hybrid were planted on May 10 and harvested on September 10 at the half milk-line and soft-dough stages, respectively. The fermentation was evaluated at 1, 2, 3, 5, 10, 15, 20, 30, and 45 days after ensiling. Although forage yield of proso millet was lower than corn and sorghum-sudangrass hybrid, its relative feed value was greater than sorghum-sudangrass hybrid. Concentrations of dry matter (DM), crude protein, and water-soluble carbohydrate decreased commonly in the ensiling forage crops. The DM loss was greater in proso millet than those in corn and sorghum-sudangrass hybrid. The in vitro dry matter digestibility declined in the forage crops as fermentation progressed. In the early stages of fermentation, pH dropped rapidly, which was stabilized in the later stages. Compared to corn and sorghum-sudangrass hybrid, the concentration of ammonia-nitrogen was greater in proso millet. The count of lactic acid bacteria reached the maximum level on day 10, with the values of 6.96, 7.77, and 6.95 Log10 CFU/g fresh weight for proso millet, corn, and sorghum-sudangrass hybrid, respectively. As ensiling progressed, the concentrations of lactic acid and acetic acid of the three crops increased and lactic acid proportion became higher in the order of sorghum-sudangrass hybrid, corn, and proso millet. Overall, the shorter, fast-growing proso millet comparing with corn and sorghum-sudangrass hybrid makes this forage crop an alternative option, particularly in areas where agricultural inputs are limited. However, additional research is needed to evaluate the efficacy of viable strategies such as chemical additives or microbial inoculants to minimize ammonia-nitrogen formation and DM loss during ensiling.

Ineffectiveness of Skin Tests in Predicting Allergic Reactions Induced by Chinese Herbal Injections.

OBJECTIVE: To evaluate whether skin tests are suitable to predict the allergy reactions induced by Chinese herbal injections (CHIs). METHODS: The skin tests including skin prick tests (SPT), intradermal tests (IDT) and provocation tests including subcutaneous tests and intravenous tests were administered to 249 healthy subjects and 180 allergic patients for 3 CHIs, including ginkgolide injection, diterpene ginkgolide meglumine injection and Salvianolate lyophilized injection. The results of the provocation tests were used as the "gold standard" to determine the sensitivity and specificity of the skin tests. RESULTS: The results did not show any significant differences between the healthy and allergy groups in both skin tests and provocation tests (P>0.05). The specificities of SPT and IDT were 0.976 and 0.797, respectively, and the sensitivities of both SPT and IDT were 0. CONCLUSION: Skin tests are insufficient to predict the likelihood of allergic reactions resulting from CHIs. (ChiCTR-CPC-15006921).

Effects of formic acid and lactic acid bacteria inoculant on main summer crop silages in Korea.

To improve the fermentation quality of silage and reduce the nutrients loss of raw materials during the ensiling process, silage additives are widely used. The effect of additives on silage is also affected by the species of crop. Therefore, this study was designed to explore the effects of formic acid (FA) and lactic acid bacterial inoculant on the quality of main summer crop silage. The experiment was consisted on split-plot design with three replications. The experiment used the main summer forage crops of proso millet ("Geumsilchal"), silage corn ("Gwangpyeongok"), and a sorghum-sudangrass hybrid ("Turbo-gold"). Treatments included silage with Lactic acid bacterial Inoculant (Lactobacillus plantarum [LP], 1.0 × 106 CFU/g fresh matter), with FA (98%, 5 mL/kg), and a control (C, without additive). All silages were stored for 60 days after preparation. All additives significantly increased the crude protein content and in vitro dry matter digestibility (IVDMD) of the silages and also reduced the content of ammonia nitrogen (NH3-N) and pH. Corn had the highest content of IVDMD, total digestible nutrients and relative feed value among silages. Compared with the control, irrespective of whether FA or LP was added, the water soluble carbohydrate (WSC) of three crops was largely preserved and the WSC content in the proso millet treated with FA was the highest. The treatment of LP significantly increased the lactic acid content of the all silage, while the use of FA significantly increased the content of acetic acid (p < 0.05). The highest count of lactic acid bacteria (LAB) was detected in the LP treatment of corn. In all FA treatment groups, the total microorganism and mold numbers were significantly lower than those of the control and LP groups (p < 0.05). In conclusion, both additives improved the fermentation quality and nutritional composition of the main summer forage crops. The application of FA effectively inhibited the fermentation of the three crops, whereas LAB promoted fermentation. So, both FA and LP can improve the quality of various species of silage.

Changes in fermentation pattern and quality of Italian ryegrass (Lolium multiflorum Lam.) silage by wilting and inoculant treatments.

OBJECTIVE: This study was conducted to investigate the effects of wilting and microbial inoculant treatment on the fermentation pattern and quality of Italian ryegrass silage. METHODS: Italian ryegrass was harvested at heading stage and ensiled into vinyl bags (20 cm×30 cm) for 60d. Italian ryegrass was ensiled with 4 treatments (NWNA, no-wilting noadditive; NWA, no-wilting with additive; WNA, wilting no-additive; WA, wilting with additive) in 3 replications, wilting time was 5 hours and additives were treated with 106 cfu/g of Lactobacillus plantarum. The silages samples were collected at 1, 2, 3, 5, 10, 20, 30, 45, and 60 days after ensiling and analyzed for the ensiling quality and characteristics of fermentation patterns. RESULTS: Wilting treatment resulted in lower crude protein and in vitro dry matter digestibility and there were no significant differences in acid detergent fiber (ADF), total digestible nutrient (TDN), water-soluble carbohydrate (WSC), ammonia content, and pH (p>0.05). However, wilting treatment resulted in higher ADF and neutral detergent fiber content of Italian ryegrass silage (p<0.05), and the WNA treatment showed the lowest TDN and in vitro dry matter digestibility. The pH of the silage was higher in the wilting group (WNA and WA) and lower in the additive treatment group. Meanwhile, the decrease in pH occurred sharply between the 3-5th day of storage. The ammonia nitrogen content was significantly lower in the additive treatment (p<0.05), and wilting had no effect. As fermentation progressed, the lactic and acetic acid contents were increased and showed the highest content at 30 days of storage. CONCLUSION: The wilting treatment did not significantly improve the silage fermentation, but the inoculant treatment improved the fermentation patterns and quality of the silage. So, inoculation before ensiling is recommended when preparing high quality of Italian ryegrass silage, and when wilting, it is recommended to combine inoculation for making high quality silage.

Detection of EP300-ZNF384 fusion in patients with acute lymphoblastic leukemia using RNA fusion gene panel sequencing.

EP300-ZNF384 fusion is a rare recurrent cytogenetic abnormality associated with B cell acute lymphoblastic leukemia (B-ALL), which was rarely studied in Chinese patient cohort. Here, we used a customized RNA fusion gene panel to investigate gene fusions in 56 selected acute leukemia patients without conventional genetic abnormalities. Two EP300-ZNF384 fusion forms were detected in ten cases, which were in-frame fusions of EP300 exon 6 fused with exon 3 or 2 of ZNF384. The fusions led to the lack of most functional domains of EP300. We firstly reported EP300-ZNF384 fusion in a mixed-phenotype acute leukemia (MPAL) patient whose CD33 and CD13 were negative. The rest nine B-ALL patients with EP300-ZNF384 fusion expressed CD33 and/or CD13. Fifty-six percent of B-ALL patients (5/9) with EP300-ZNF384 fusion were positive with CD10. After the diagnosis of EP300-ZNF384 fusion, 70% of the patients achieved remission after chemotherapy. Our observations indicated that EP300-ZNF384 fusion consists of a distinct subgroup of B-ALL with a characteristic immunophenotype. These patients are sensitive to current chemotherapy regimen and have an excellent outcome.

Acidic leucine-rich nuclear phosphoprotein-32A expression contributes to adverse outcome in acute myeloid leukemia.

BACKGROUND: Acidic leucine-rich nuclear phosphoprotein-32A (ANP32A) is a novel regulator of histone H3 acetylation and promotes leukemogenesis in acute myeloid leukemia (AML). However, its prognostic value in AML remains unclear. METHODS: In this study, we evaluated the prognostic significance of ANP32A expression using two independent large cohorts of cytogenetically normal AML (CN-AML) patients. Multivariable analysis in CN-AML group was also presented. Based on the ANP32A expression, its related genes, dysregulation of pathways, interaction network analysis between microRNAs and target genes, as well as methylation analysis were performed to unveil the complex functions behind ANP32A. RESULTS: Here we demonstrated overexpression of ANP32A was notably associated with unfavorable outcome in two independent cohorts of CN-AML patients (OS: P=0.012, EFS: P=0.005, n=185; OS: P=0.041, n=232), as well as in European Leukemia Net (ELN) Intermediate-I group (OS: P=0.018, EFS: P=0.045, n=115), National Comprehensive Cancer Network (NCCN) Intermediate Risk AML group (OS: P=0.048, EFS: P=0.039, n=225), and non-M3 AML group (OS: P=0.034, EFS: P=0.011, n=435). Multivariable analysis further validated ANP32A as a high-risk factor in CN-AML group. Multi-omics analysis presented overexpression of ANP32A was associated with aberrant expression of oncogenes and tumor suppressor, up/down-regulation of metabolic and immune-related pathways, dysregulation of microRNAs, and hypomethylation on CpG island and 1st Exon regions. CONCLUSIONS: We proved ANP32A as a novel, potential unfavorable prognosticator and therapeutic target for AML.

[Analysis of Clinical Characteristics of Acute B Lymphoblastic Leukemia with EP300-ZNF384 Fusion Gene Positive].

OBJECTIVE: To investigate the clinical manifestations and laboratory features of B-ALL patients with EP300-ZNF384 fusion gene positive, so as to improve the understanding of this subtype disease. METHODS: The clinical data of 3 B-ALL patients with EP300-ZNF384 fusion gene positive admitted in Department of Hematology, the first medical center of Chinese PLA general hospital from February 2017 to February 2018 were collected and analyzed retrospectively. The clinical and laboratory characteristics as well as the therapentic outcome in B-ALL patients with EP300-ZNF384 fusion gene positive were analyzed. RESULTS: The fusion gene of EP300-ZNF384 was detected in 8.1%(3/37) of B-ALL patients. All cases showed the normal karyotype and aberrant CD13 and/or CD33 expression for immunophenotype. 3 patients were sensitive to traditional chemotherapy. CONCLUSION: The B-ALL with EEP300-ZNF384 fusion gene positive may be a subgroup of B-ALL with a uniqe clinical characteristis and laboatorial features. EP300-ZNF384 positive patients show a good response to conventional chemotherapy, suggesting a favorable prognosis.

Comparison of intramuscular and intravenous pharmacokinetics of ginsenosides in humans after dosing XueShuanTong, a lyophilized extract of Panax notoginseng roots.

ETHNOPHARMACOLOGICAL RELEVANCE: Many bioactive constituents of Chinese herbal medicines have poor oral bioavailability. Besides oral administration, herbal medicines in China are also prepared for parenteral administration. Unlike for intravenous route, little is known about the intramuscular pharmacokinetics of herbal compounds. To facilitate rational use of herbal medicine, it is important to better understand such intramuscular pharmacokinetics. AIM OF THE STUDY: Bioactive constituents of XueShuanTong (a lyophilized extract of Panax notoginseng roots, extensively used in treatment of ischemic heart and cerebrovascular diseases) predominantly comprise ginsenosides Rb1 and Rd of 20(S)-protopanaxadiol-type and ginsenosides Rg1, and Re, and notoginsenoside R1 of 20(S)-protopanaxatriol-type; these saponins are poorly absorbed from the gastrointestinal tract. This study aimed to compare intramuscular and intravenous pharmacokinetics of these ginsenosides after dosing XueShuanTong. METHODS: Pharmacokinetics of ginsenosides was assessed in human volunteers receiving an intramuscular injection or 1.5-h intravenous infusion of XueShuanTong, both at 150 mg/person, and the plasma and urine samples were analyzed by liquid chromatography/mass spectrometry. RESULTS: Like after intravenous administration, the unchanged saponins were the major circulating forms after intramuscular administration, while their metabolites were poorly detected. These ginsenosides exhibited intramuscular bioavailability of 100%-112%, relative to the respective intravenous data. Similar to that after intravenous infusion, the 20(S)-protopanaxadiol-type ginsenosides after the intramuscular injection exhibited notably longer terminal half-lives (46-106 h) than the 20(S)-protopanaxatriol-type ginsenosides (1.1-1.4 h). CONCLUSIONS: Intramuscular route might be an effective alternative to intravenous route for XueShuanTong, from the pharmacokinetic perspective.

[Clinical Retrospective Analysis of Chronic Myeloid Leukemia in Myeloid Blast Crisis].

OBJECTIVE: To retrospectively analyze the clinical manifestation, laboratorial test features and prognosis of patients with CML in myeloid blast crisis. METHODS: The clinical data of 10 patients with CML in myeloid blast crisis admitted in Chinese PLA General Hospital from June 2011 to May 2018 were collected, and their clinical features, laboratorial data and long-term survival were analyzed. RESULTS: The median age of these 10 cases was 32.5 (23-73) years old. Nine cases had chronic phase history. The median chronic phase was 17(4-84) months. All the 10 cases had splenomegaly; B-ultrasonography showed that the median spleen size was 5.2 (4-7.8) cm in thickness, and 14.6 (11.4-19.8) in length. When chronic myeloid leukemia was in blast crisis, the median WBC count was 41.705（11.9-344.41）×109 /L and the median platelet count was 159 (13-2326) ×109 /L. The Ph+ chromosome and BCR-ABL1 fusion gene coulld be detected in all the cases. The chromosome karyotyping showed that additional chromosome abnormalities were found in 5 cases. One case was of low diploid, and two cases were with complex karyotype. ABL1 mutation was detected in 6 out of these 10 cases. ABL1 T315I mutation was detected in 2 of them and one was with deletion of combined P53 in genetic tests. The median follow-up time was 10.5(0.2-78) months. There were 5 cases treated sequentially by chemotheraphy with or without TKI and allo-HSCT. Three cases reached CP2 before transplantation. Among them, two cases still survived without progression for 67 months and 69 months after the transplantation respectively. One case died of transplantation-related mortality (suffered from cerebral hemorrhage 7 months after the transplantation). Two cases were NR before the transplantation, and both died of disease relapse or progression at the time points of one or three months after the transplantation. Five cases treated by TKI ± chemotheraphy and without HSCT succumbed to disease progression. The median time was 6(0.2-22) months. CONCLUSION: CML patients in myeloid blast crisis treated by chemotheraphy combined with TKI gain CP2, the survival time of patients treated by sequential allo-HSCT is prolonged.

Intravenous formulation of Panax notoginseng root extract: human pharmacokinetics of ginsenosides and potential for perpetrating drug interactions.

XueShuanTong, a lyophilized extract of Panax notoginseng roots (Sanqi) for intravenous administration, is extensively used as add-on therapy in the treatment of ischemic heart and cerebrovascular diseases and comprises therapeutically active ginsenosides. Potential for XueShuanTong-drug interactions was determined; the investigation focused on cytochrome P450 (CYP)3A induction and organic anion-transporting polypeptide (OATP)1B inhibition. Ginsenosides considerably bioavailable for drug interactions were identified by dosing XueShuanTong in human subjects and their interaction-related pharmacokinetics were determined. The CYP3A induction potential was determined by repeatedly dosing XueShuanTong for 15 days in human subjects and by treating cryopreserved human hepatocytes with circulating ginsenosides; midazolam served as a probe substrate. Joint inhibition of OATP1B by XueShuanTong ginsenosides was assessed in vitro, and the data were processed using the Chou-Talalay method. Samples were analyzed by liquid chromatography/mass spectrometry. Ginsenosides Rb1, Rd, and Rg1 and notoginsenoside R1 were the major circulating XueShuanTong compounds; their interaction-related pharmacokinetics comprised compound dose-dependent levels of systemic exposure and, for ginsenosides Rb1 and Rd, long terminal half-lives (32‒57 and 58‒307 h, respectively) and low unbound fractions in plasma (0.8%‒2.9% and 0.4%‒3.0%, respectively). Dosing XueShuanTong did not induce CYP3A. Based on the pharmacokinetics and inhibitory potency of the ginsenosides, XueShuanTong was predicted to have high potential for OATP1B3-mediated drug interactions (attributed chiefly to ginsenoside Rb1) suggesting the need for further model-based determination of the interaction potential for XueShuanTong and, if necessary, a clinical drug interaction study. Increased awareness of ginsenosides' pharmacokinetics and XueShuanTong-drug interaction potential will help ensure the safe use of XueShuanTong and coadministered synthetic drugs.

Inhibitory effects of lappaconitine on the neuronal isoforms of voltage-gated sodium channels.

Lappaconitine (LA) has been widely used for postoperative and cancer pain control. LA exhibits excellent analgesic activity with a longer effective time than common local anesthetics such as tetracaine and bupivacaine. However, the mechanisms underlying the featured analgesic activity of LA remain largely unknown. Here, we report that LA is an inhibitor of voltage-gated sodium channel 1.7 (Nav1.7) stably expressed in human embryonic kidney (HEK293) cells. LA inhibited Nav1.7 in a voltage-dependent manner with an IC50 value (with 95% confidence limits) of 27.67 (15.68-39.66) µmol/L when the cell was clamped at -70 mV. In comparison with the quick and reversible inhibition of Nav1.7 by tetracaine and bupivacaine, the inhibitory effect of LA was rather slow and irreversible. It took more than 10 min to achieve steady-state inhibition when LA (300 µmol/L) was administered. Unlike tetracaine and bupivacaine, LA affected neither the voltage-dependent activation nor the inactivation of the channels. Five residues in domain III and domain IV have been reported to be critical for the effects of the two local anesthetics on Nav channels. But our mutant study revealed that only two residues (F1737, N1742) located in domain IV were necessary for the inhibitory activity of LA. The slow onset, irreversibility, and lack of influence on channel activation and inactivation accompanied with the different molecular determinants suggest that LA may inhibit Nav1.7 channels in a manner different from local anesthetics. These results may help to understand the featured analgesic activity of LA, thus benefiting its application in the clinic and future drug development.

Human pharmacokinetics of ginkgo terpene lactones and impact of carboxylation in blood on their platelet-activating factor antagonistic activity.

Terpene lactones are a class of bioactive constituents of standardized preparations of Ginkgo biloba leaf extract, extensively used as add-on therapies in patients with ischemic cardiovascular and cerebrovascular diseases. This investigation evaluated human pharmacokinetics of ginkgo terpene lactones and impact of their carboxylation in blood. Human subjects received oral YinXing-TongZhi tablet or intravenous ShuXueNing, two standardized ginkgo preparations. Their plasma protein-binding and platelet-activating factor antagonistic activity were assessed in vitro. Their carboxylation was assessed in phosphate-buffered saline (pH 7.4) and in human plasma. After dosing YinXing-TongZhi tablet, ginkgolides A and B and bilobalide exhibited significantly higher systemic exposure levels than ginkgolides C and J; after dosing ShuXueNing, ginkgolides A, B, C, and J exhibited high exposure levels. The compounds' unbound fractions in plasma were 45-92%. Apparent oral bioavailability of ginkgolides A and B was mostly >100%, while that of ginkgolides C and J was 6-15%. Bilobalide's bioavailability was probably high but lower than that of ginkgolides A/B. Terminal half-lives of ginkgolides A, B, and C (4-7 h) after dosing ShuXueNing were shorter than their respective values (6-13 h) after dosing YinXing-TongZhi tablet. Half-life of bilobalide after dosing the tablet was around 5 h. Terpene lactones were roughly evenly distributed in various body fluids and tissues; glomerular-filtration-based renal excretion was the predominant elimination route for the ginkgolides and a major route for bilobalide. Terpene lactones circulated as trilactones and monocarboxylates. Carboxylation reduced platelet-activating factor antagonistic activity of ginkgolides A, B, and C. Ginkgolide J, bilobalide, and ginkgo flavonoids exhibited no such bioactivity. Collectively, differences in terpene lactones' exposure between the two preparations and influence of their carboxylation in blood should be considered in investigating the relative contributions of terpene lactones to ginkgo preparations' therapeutic effects. The results here will inform rational clinical use of ginkgo preparations.

Enhancing inactivation rather than reducing activation of Nav1.7 channels by a clinically effective analgesic CNV1014802.

The Nav1.7 channel represents a promising target for pain relief. In the recent decades, a number of Nav1.7 channel inhibitors have been developed. According to the effects on channel kinetics, these inhibitors could be divided into two major classes: reducing activation or enhancing inactivation. To date, however, only several inhibitors have moved forward into phase 2 clinical trials and most of them display a less than ideal analgesic efficacy, thus intensifying the controversy regarding if an ideal candidate should preferentially affect the activation or inactivation state. In the present study, we investigated the action mechanisms of a recently clinically confirmed inhibitor CNV1014802 using both electrophysiology and site-directed mutagenesis. We found that CNV1014802 inhibited Nav1.7 channels through stabilizing a nonconductive inactivated state. When the cells expressing Nav1.7 channels were hold at 70 mV or 120 mV, the half maximal inhibitory concentration (IC50) values (with 95% confidence limits) were 1.77 (1.20-2.33) and 71.66 (46.85-96.48) µmol/L, respectively. This drug caused dramatic hyperpolarizing shift of channel inactivation but did not affect activation. Moreover, CNV1014802 accelerated the onset of inactivation and delayed the recovery from inactivation. Notably, application of CNV1014802 (30 µmol/L) could rescue the Nav1.7 mutations expressed in CHO cells that cause paroxysmal extreme pain disorder (PEPD), thereby restoring the impaired inactivation to those of the wild-type channel. Our study demonstrates that CNV1014802 enhances the inactivation but does not reduce the activation of Nav1.7 channels, suggesting that identifying inhibitors that preferentially affect inactivation is a promising approach for developing drugs targeting Nav1.7.

[Effects of Simvastatin on Expression of Multidrug Resistance Gene and Protein of SHI-1 Cells].

OBJECTIVE: To investigate the effects of simvastatin(SIM) and serum free medium(SFM) on the expression of multidrug resistance gene(MDR1) and protein of SHI-1 cells. METHODS: Trypan blue exclusion assay was used to detect the proliferation level and viability of SHI-1 cells after treatment with SIM and culture in SFM; The multi-drug resistant protein p-gp was measured by flow cytometry after culture in SFM for 1 to 3 days and treatment with various concentration of simvastatin. The effect of SFM culture and SIM treatment on the expression of MDR1 trascript was detected by qPCR; ELISA was used to measure the change of cellular cholesterol after culture in SIM and SFM. Chemosensitivity assay was performed after treatment with SIM for SHI-1 cells. RESULTS: Compared with control group, the growth of SHI-1 cells cultured in SFM decreased in a time-dependent manner. The growth-inhibitory effect was markedly increased when SHI-1 cells were treated with SIM and SFM. The mRNA level of MDR1 gene decreased after SIM treatment or/and culture in SFM. P-gp protein was downregulated in SHI-1 cells cultured in SFM or/and treated with SIM. The cellular cholesterol level increased when the cells were cultured in SFM. Total cellular cholesterol level decreased in SHI-1 cells treated with SIM and cultured in SFM. Chemosensitivity assays found that pre-treatment with SIM could increase the cytotoxicity of DNR to SHI-1 cells. CONCLUSION: Culture with SIM and SFM can downregulate the expression of MDR1 gene and p-gp protein in SHI-1 cells. SIM also can enhance the chemotherapeutic sensitivity of SHI-1 cells.