Meranzin Hydrate Improves Depression-Like Behaviors and Hypomotility via Ghrelin and Neurocircuitry / 中国结合医学杂志

OBJECTIVE@#To investigate whether meranzin hydrate (MH) can alleviate depression-like behavior and hypomotility similar to Chaihu Shugan Powder (CSP), and further explore the potential common mechanisms.@*METHODS@#Totally 120 Spraque-Dawley rats were randomly divided into 5-8 groups including sham, vehicle, fluoxetine (20 mg/kg), mosapride (10 mg/kg), CSP (30 g/kg), MH (9.18 mg/kg), [D-Lys3]-GHRP-6 (Dlys, 0.5 mg/kg), and MH+Dlys groups by a random number table, 8 rats in each group. And 32 mice were randomly divided into wild-type, MH (18 mg/kg), growth hormone secretagogue receptor-knockout (GHSR-KO), and GHSR+MH groups, 8 mice in each group. The forced swimming test (FST), open field test (OFT), tail suspension test (TST), gastric emptying (GE) test, and intestinal transit (IT) test were used to assess antidepressant and prokinetic (AP) effects after drug single administration for 30 min with absorbable identification in rats and mice, respectively. The protein expression levels of brain-derived neurotrophic factor (BDNF) and phosphorylated mammalian target of rapamycin (p-mTOR) in the hippocampus of rats were evaluated by Western blot. The differences in functional brain changes were determined via 7.0 T functional magnetic resonance imaging-blood oxygen level-dependent (fMRI-BOLD).@*RESULTS@#MH treatment improved depression-like behavior (FST, OFT) and hypomotility (GE, IT) in the acute forced swimming (FS) rats (all P<0.05), and the effects are similar to the parent formula CSP. The ghrelin antagonist [D-Lys3]-GHRP-6 inhibited the effect of MH on FST and GE (P<0.05). Similarly, MH treatment also alleviated depression-like behavior (FST, TST) in the wild-type mice, however, no effects were found in the GHSR KO mice. Additionally, administration of MH significantly stimulated BDNF and p-mTOR protein expressions in the hippocampus (both P<0.01), which were also prevented by [D-Lys3]-GHRP-6 (P<0.01). Besides, 3 main BOLD foci following acute FS rats implicated activity in hippocampus-thalamus-basal ganglia (HTB) circuits. The [D-Lys3]-GHRP-6 synchronously inhibited BOLD HTB foci. As expected, prokinetic mosapride only had effects on the thalamus and basal ganglia, but not on the hippocampus. Within the HTB, the hippocampus is implicated in depression and FD.@*CONCLUSIONS@#MH accounts for part of AP effects of parent formula CSP in acute FS rats, mainly via ghrelin-related shared regulation coupled to BOLD signals in brain areas. This novel functionally connection of HTB following acute stress, treatment, and regulation highlights anti-depression unified theory.

Improvement on Quality Standard of Belladonnae Herba / 中国实验方剂学杂志

ObjectiveTo improve the current standard of Belladonnae Herba in the 2020 edition of Chinese Pharmacopoeia. MethodTaking hyoscyamine sulfate， atropine sulfate and scopoletin as reference substances， and ethyl acetate-methanol-concentrated ammonia（17∶4∶2）as developing solvent， thin layer chromatography （TLC） was applied in the qualitative identification of Belladonnae Herba. The moisture， total ash and ethanol-soluble extract of Belladonnae Herba were determined based on the general principles in the 2020 edition of Chinese Pharmacopoeia （volume Ⅳ）. The contents of hyoscyamine sulfate and scopolamine hydrobromide were analyzed by high performance liquid chromatography （HPLC） with mobile phase of acetonitrile-54 mmol·L-1 phosphate buffer solution （14∶86）， flow rate of 1.0 mL·min-1 and detection wavelength at 210 nm. ResultThe spots in the TLC were clear with good separation and specificity. Hyoscyamine sulfate and scopolamine hydrobromide showed a good linearity with peak area in the range of 0.024 7-0.789 6 g·L-1 （r=0.999 9） and 0.003 9-0.124 0 g·L-1 （r=0.999 9）， the average recoveries of these two ingredients were 100.29% （RSD 1.6%） and 99.04% （RSD 1.4%）， respectively. The limits for moisture， total ash in Belladonnae Herba should be less than 13.0% and the limit for the ethanol-soluble extract should be more than 10.0%. Due to the low content and wide variation of scopolamine hydrobromide， the content of hyoscyamine sulfate should not be less than 0.098%. ConclusionThe established method is simple， specific and reproducible， which can be used to improve the quality control standard of Belladonnae Herba.

Studies on chemical constituents of Isodon henryi / 中国中药杂志

The chemical constituents of the water extraction of the aerial parts of Isodon henryi were investigated by various chromatographic methods including D-101 macroporous adsorptive resins,silica gel,sephadex LH-20,and semi-preparative HPLC. As a result,ten compounds were separated and purified. By analyses of the UV,IR,MS,NMR spectra,their structures were determined as rabdosinate( 1),lasiokaurin( 2),epinodosinol( 3),rabdosichuanin C( 4),epinodosin( 5),hebeirubescensin k( 6),rubescensin C( 7),enmenol( 8),oridonin( 9),and enmenol-1-β-glucoside( 10). Compounds 1-8 and 10 were isolated from I. henryi for the first time. Compounds 2 and 9 showed inhibitory effects against four tumor cells,with IC50 values of 2. 25-9. 32 μmol·L-1.

Significance of Bone Marrow Unclassifiable Cells in Diagnosis of Fever of Unknown Origin / 中国实验血液学杂志

OBJECTIVE@#To investigate the clinical significance of bone marrow unclassifiable cells in diagnosis of fever of unknown origin(FUO).@*METHODS@#The clinical data of 60 patients with FUO admitted in the first affiliated hospital of Xi'an Jiaotong university from June 2014 to May 2016 were collected, and 60 patients with FUO were divided into 2 group: group A(30 cases) in which the unclassifiable cells in bone marrow were observed by bone marrow examination, and group B(30 cases) in which the unclassifiable cells in bone marrow not were found by bone marrow examination. The clinical characteristics, bone marrow features, immunophenotypes of bone marrow cells and prognosis of patients in 2 groups were analyzed retrospectively.@*RESULTS@#Out of 30 patients in group A, 18 were diagnosed as malignant tumors including 12 cases of lymphoma, while out of 30 patients in group B, 5 cases were diagnosed as malignant tumor, including 3 cases of lymphoma. For the patients with non-tumor diseases, the bone marrow unclassifiable cells disappeared after the patients condition was improved.@*CONCLUSION@#The bone marrow examination including the smear and biopsy shonld be performed routinely for the patients with FUO. If the unclassifiable cells are observed morphologically in bone marrow of patients with FUO, the disease of patients should be considered as malignant tumor, especially, lymphoma.

Cloning and Expression Analysis of PaAACT Gene from Phytolacca americana / 中国实验方剂学杂志

Objective: To clone the key enzyme gene involved in the biosynthesis of esculentoside A(EsA),acetoacetyl-CoA transferase(AACT) gene was cloned from Phytolacca americana for bioinformatics analysis and prokaryotic expression. Method: Total RNA was extracted from the root of P. americana, and then cDNA was synthesized through the reverse transcription. Based on analysis of the transcriptome data of P. americana, the specific primers of PaAACT gene were designed,and the cDNA sequence of PaAACT gene was amplified by polymerase chain reaction(PCR) method. Prokaryotic induction,expression and purification of the target protein were induced through the construction of the prokaryotic expression vector pET-32a-PaAACT. Result: The open reading frame (ORF) of PaAACT gene was 1 254 bp,and encoded 417 amino acid residues. Bioinformatic analysis showed that the molecular formula of PaAACT protein was C1 914H3 120N538O576S17,inferring that its molecular weight was 43.43 kDa,the theoretical isoelectric point was 8.90,and the instability index of PaAACT protein was 32.27,which was a stable protein. According to bioinformatics analysis,PaAACT protein was a member of the thiolase family and contained one conserved site and one active site of the thiolase family at the C-terminal. PaAACT protein may be located in the cytoplasm,without a signal peptide or transmembrane domain. The phylogenetic analysis indicated that PaAACT protein showed the highest homology with AACT protein from polygonaceae plants (such as Beta vulgaris). The recombinant PaAACT protein was successfully expressed in Escherichia coli BL21(DE3) strain through IPTG induction, and the purified target protein was obtained by Ni2+ affinity chromatography. Conclusion: In this study,the PaAACT gene was cloned from P. americana,which lays a foundation for further determination of enzyme activity assay of PaAACT and preparation of antibody,and provides the theoretical basis for studying its role in the biosynthesis pathway of EsA.

Significance of Simultaneous Analysis of Bone Marrow Smear and Bone Marrow Biopsy in the Diagnosis of Multiple Myloma / 中国实验血液学杂志

<p><b>OBJECTIVE</b>To explore the value of bone marrow smear and biopsy simultaneously applied to diagnosis of multiple myloma (MM).</p><p><b>METHODS</b>Clinical data of 30 cases of multiple myloma were collected from our hospital in the year 2014 and analyzed retrospectively, and the results of the bone marrow smear and the simultaneous bone marrow biopsy were compared.</p><p><b>RESULTS</b>Hyperplasia levels in bone marrow biopsy was significantly higher than that in bone marrow smears, and the active and highly active hyperplasia of nucleated cells were observed in all the bone marrow biopsies; the myeloma cells showed a focal or diffuse distribution, the binuclear or polynuclear myeloma cells were observed in 22 patients (73%), but the detection rate of abnormal myeloma cells was 40% in bone marrow smear (P < 0.05). There was mild to moderate hyperplasia of fibrous tissue in bone marrow biopsy, and the hyperplasia degeree of fibrous tissue strongly positively correlated with the myeloma cell ratio (r = 0.412).</p><p><b>CONCLUSION</b>The bone marrow smear and aspiration biopsy can complement each other so as to reduce the misdiagnosis rate, therefore contributes to the early diaglosis and treatment.</p>

Pathological Characteristics of Bone Marrow in Non-Hodgkin's Lymphoma Patients with Secondary Myelofibrosis and Their Relationship with Prognosis / 中国实验血液学杂志

<p><b>OBJECTIVE</b>To investigate the pathological characteristics of bone marrow in non-Hodgkin's lymphoma(NHL) patients with secondary myelofibrosis and their relationship with disease prognosis.</p><p><b>METHODS</b>The pathological characteristics of bone marrow in 14 NHL patients with secondary myelofibrosis and 30 NHL patients without secondary myelofibrosis received from January 2012 to December 2013 in Department of Hematology, the First Affiliated Hospital Xi'an Jiaotong University Medical School were analysed, and overall survival and progress-free survival rates of NHL patients with and without secondary myelofibrosis were analyzed and compared.</p><p><b>RESULTS</b>14 cases of NHL with secondary myelofibrosis including 9 cases of lymphoma cell leukemia were all at stage IV and had hyperplasia of bone marrow fibrous tissue, the Gomori staining were all positive. When the lymphoma cells in bone marrow decreased or negative, their Gomori staining were negative. If the disease relapsed, the Gomori staining became positive again. There were 30 cases of NHL at stage IV wihtout secondary myelofibrosis. The overall survival rates and progression-free survival rates at 1,3,5 years in these patients were 100%, 93.1%, 57.6% and 100%, 92.6%,52.6% respectively. The overall survival rates and progression-free survival rates at 1,3,5 years in 14 NHL patients with secondary myelofibrosis were 92.9%,81.3%, 48.8% and was 71.8%, 62.3%, 47.9%, respectively.</p><p><b>CONCLUSION</b>NHL patients with secondary myelofibrosis are almost at stage IV, especially in the patients with lymphoma cell leukemia. They had different degree of hyperplasia of bone marrow fibrous tissue, and the myelofibrosis would be reduced or disappeared when the disease in remission. The overall and progression -free survival rates decrease in NHL patients with secondary myelofibrosis, compared with patients without secondary myelofibrosis. Secondary myelofibrosis is one of the indicators of poor prognosis.</p>

Differential analysis of BM cell morphology, immunophenotypic, cytogenetic characters and prognosis between myeloblastic and lymphoblastic crisis of CML / 中国实验血液学杂志

This study was purposed to investigate the difference of morphology, immunophenotype, cytogenetic features and prognosis between myeloid blast crisis and lymphoid blast crisis of chronic myelogenous leukemia (CML). A total of 31 patients with CML in blastic crisis in Department of Hematology, the First Affiliated Hospital of Xi'an Jiaotong University school of Medicine from 2009 January to 2014 January were enrolled in this study. Out of 31 CML patients, 24 cases were patients with myeloid blast crisis and other 7 cases were patients with lymphoblastic crisis. The clinical data, blast cell percentage in peripheral blood and bone marrow, eosinophil and basophil percentage, immunophenotype, cytogenetic characteristics and prognosis were analyzed. The results indicated that there was no significant difference of blastic cell percentage in peripheral blood and bone marrow of CML with myeloid blast crisis, and the eosinophil and basophil cells could be easily detected. The ratio of blastic cells in BM was higher than that in PB in lymphoid blastic crisis of CML, eosinophil and basophil cells were rare. 7 cases of CML with lymphoid blastic crisis were B ALL with CD10, CD19, CD34, HLA-DR expression, and 2 cases with CD13 and CD33 expression. The lymphoid score was in all CML patients with lymphoid blastic crisis was greater than or equal to 1.5;and 2 patients with CD13 and CD33 expression, and with 1 myeloid score.24 cases of myeloid blastic crisis of CML patients mainly expressed CD33, CD13, CD38, CD34, CD11b and HLA-DR, and their myeloid score greater than or equal to 2, among them the lymphoid scores of 2 patients were 0.5 and 1 score, respectively. All the 31 patients showed 100% Ph(+) chromosome, among them 3 cases also showed other new chromosome aberrations. There was no significant difference of overall survival rate between lymphoid and myeloid blastic crisis of CML, but the overall survival rate of patients treated with tyrosine kinase inhibitor (TKI ) was higher than that in the patients without TKI treatment. It is concluded that eosinophil and basophil cells in peripheral blood of lymphoid blastic crisis were less than that of CML patients with myeloid blastic crisis. Lymphoid blastic crisis of CML patients occurred mostly in B ALL cases with expression of CD10 and CD19. Patients with myeloid blastic crisis of CML mainly expressed CD33, CD13, CD38, CD34, CD11b and HLA-DR, and could be accompanied by other lineage antigen expression, but the score was less than 2. New chromosome aberration is easily observed in myeloid blastic crisis of CML. There is no significant difference of overall survival rate of between CML patients with lymphoid and myeloid blastic crisis, but the overall survival rate of patients treated with TKI is higher than the patients without TKI treatment.

Analysis of bone marrow and peripheral blood cytologic features in hyperleukocytic acute leukemia / 中国实验血液学杂志

This study was purpose to investigate the cytologic features of bone marrow (BM) and peripheral blood (PB) in hyperleukocytic acute leukemia (HAL) and their clinical significance in accordance with high leukocyte count as poor prognostic factor for acute leukemia. The smears of BM and PB were collected from 68 out-patients and inpatients including 28 cases of HLA and 40 cases of non HAL (NHAL) in our hospital since 2009. The proliferation degree, morphology and abnormal appearance in each cell lineage were observed with HE, POX, PAS, NSE+ NaF staining for BM mears and HE staining for PB smears by means of optical microscope. The final diagnosis was made by cellular chemical staining results, then the counting and classification were performed in 200 nucleated cells to calculate the percentage of each cell lineage, the myeloid/erythroid ratio and so on. The BP smears were observed with the same methods, the counting and classification of 100 nucleated cells were performed to calculate a variety of nucleated cell percentage. The resulted data were analyzed by the SPSS 12.0 statistical software, the difference of proliferation degree and ratio of each cell lineage in BM smears were compared, the relationship of morphological features of PB smears with BM smears was analyzed. The results showed that obvious or extreme active proliferation of nucleated cells was observed in HAL and NHAL groups, but the myeloproliferation in HAL group was more active than that in NHAL group (P < 0.05). The erythrocyte and megakaryocyte lineages were suppressed in both groups, while the HAL group showed a lower proportion of erythrocyte and megakaryocyte lineages in BM as compared with NHAL group (P < 0.05). The hemoglobin and platelet levels in PB of HAL group were obviously lower than those in PB of NHAL group (P < 0.05). The leukemia cells could be seen in PB smears of NHAL, but the proportion of leukemia cells in NHAL group was smaller than that in HAL group (P < 0.05). The leukocyte count in PB of HAL group strongly positively correlated with the proliferation degree of leukemia cells in BM of HAL group (r = 0.422). It is concluded that the significant difference of proliferation degree, cell levels and blast ratio in BM and PB exists in HAL and NHAL groups, moreover the leukemia cells ratio, leukocyte, hemoglobin and platelet levels in PB of HAL all show characteristic changes. Therefore the contrast analysis of characteristic changes from laboratorial detection contributes to grasp the regular pattern of HAL, meanwhile has an important value for guiding correct diagnosis of acute leukemia and choosing suitable treatment options.

In vitro anti-myeloma effects induced by myeloma idiotype-protein pulsed dendritic cell vaccine / 中华血液学杂志

<p><b>OBJECTIVE</b>To investigate the specific antitumor immune response induced by idiotype protein (Id)-pulsed dendritic cells (DC) in vitro.</p><p><b>METHODS</b>DC was generated from peripheral blood monocytes of the multiple myeloma (MM) patients using GM-CSF, IL-4, and TNF-alpha. The DCs were pulsed with idiotypic fragment, the F(ab')2 fragment of M protein from MM patient at the immature stage. The morphologic characteristics of the cells were observed with light and electron microscopes. The phenotypic features were analyzed with FACS, MTT assay was employed to evaluate the proliferation of autologous T cells and the inhibition rate of MM cells.</p><p><b>RESULTS</b>DC precursors in peripheral blood could be induced to typical mature DC in medium containing GM-CSF, IL-4 and TNF-alpha. Mature DC with Id could increase the proliferation of the autologous T cells and activate naive T cells to become tumor specialized cytotoxic T lymphocytes (CTL). The CTL at different doses showed significant inhibition on or killing ability to autologous MM cells in vitro.</p><p><b>CONCLUSIONS</b>In a suitable cytokine environment, the DC precursors from peripheral blood of MM patients could be induced to functional DC, and vaccination of Id-pulsed DC could induce active antitumor immune response.</p>