Research progress on chemical constituents from Chloranthus plants and their biological activities / 中国中药杂志

The genus Chloranthus has 13 species and 5 varieties in China, which can be found in the southwest and northeast regions. Phytochemical studies on Chloranthus plants have reported a large amount of terpenoids, such as diterpenoids, sesquiterpenoids, and sesquiterpenoid dimers. Their anti-inflammation, anti-tumor, antifungal, antivirus, and neuroprotection activities have been confirmed by previous pharmacological research. Herein, research on the chemical constituents from Chloranthus plants and their biological activities over the five years was summarized to provide scientific basis for the further development and utilization of Chloranthus plants.

Chemical constituents of sesquiterpenes from Chloranthus multistachys / 中国中药杂志

With repeated silica gel, octadecyl silica(ODS), and Sephadex LH-20 column chromatography, normal-phase and reverse-phase high performance liquid chromatography(HPLC), etc., a pair of new enantiomers and 5 known compounds were separated from the 95% ethanol extract of Chloranthus multistachys. These compounds were identified by the nuclear magnetic resonance spectroscopy(including 1 D-NMR and 2 D-NMR), single-crystal X-ray diffraction, circular dichroism(CD) spectroscopy, mass spectrometry(MS), and some other methods as(1R,4R,5R,8S,10R)-chloraeudolide H(1 a),(1S,4S,5S,8R,10S)-chloraeudolide H(1 b), hydroxyisogermafurenolide(2), 4α-hydroxy-5α,8β(H)-eudesm-7(11)-en-8,12-olide(3), chloraniolide A(4), chlorantene D(5), 4α,8β-dihydroxy-5α(H)-eudesm-7(11)-en-8,12-olide(6). Compounds 1 a and 1 b are a pair of new eudesmane-type sesquiterpene enantiomers, and compounds 2-4 were isolated from C. multistachys for the first time.

Therapeutic Effect of PE+CRRT Combined with Chemotherapy on Children with Severe EBV-HLH and Non-EBV-HLH / 中国实验血液学杂志

OBJECTIVE@#To investigate the difference in the therapeutic effect of plasma exchange and continuous renal replacement therapy (PE+CRRT) combined with chemotherapy in the treatment of children with severe Epstein-Barr virus-associated hemophagocytic lymphohistiocytosis (EBV-HLH) and non-EBV-HLH.@*METHODS@#The clinical data of 21 cases of all children with severe HLH treated by PE+CRRT combined with chemotherapy from January 2017 to January 2020 were collected and retrospectively analyzed. According to the presence of EBV infection, the children were divided into EBV@*RESULTS@#Among the 21 children, 14 were divided into the EBV@*CONCLUSION@#PE+CRRT combined with chemotherapy can reduce serum ferritin quickly, then improve organ function, and increase the overall survival rate of severe HLH, and it is a good effect on children with severe EBV-HLH and non-EBV-HLH.

A new sesquiterpene from Chloranthus henryi / 中国中药杂志

Eight sesquiterpenes were isolated and purified from the ethanol extract of Chloranthus henryi by column chromatographies over silica gel, ODS and Sephadex LH-20,and preparative HPLC. Their chemical structures were established by spectral data and physiochemical properties as(1S,6S,8S,10R)-8-ethoxy-10-methoxychlomultin C(1),tianmushanol(2),multistalide A(3),myrrhterpenoid N(4),1α,9α-dihydroxy-8,12-expoxy-eudesma-4,7,11-trien-6-one(5),4β,10α-aromadendranediol(6),oplopanone(7),10α-hydroxycadinan-4-en-3-one(8). Among them, compound(1) was a new compound, and compounds 2-8 were isolated from Chloranthus henryi for the first time.

A pair of enantiomeric sesquiterpenoids with neuroprotective effects from Chloranthus henryi / 药学学报

An ethanol extract of Chloranthus henryi (Chloranthaceae) was subjected to various chromatographic procedures including silica gel column chromatography, MCI column chromatography, Sephadex LH-20 column chromatography, and preparative HPLC. Five purified sesquiterpenes analyzed by spectroscopic analyses (MS, IR, NMR) and single crystal X-ray diffraction were elucidated as (1S,6S,8R)-8-ethoxychlomultin C (1a), (1R,6R,8S)-8-ethoxychlomultin C (1b), (+)-phaeocaulin D (2), atractylenolide Ⅰ (3), and 8-β-ethoxyasterolid (4). Compounds 1a and 1b were a new pair of sesquiterpene enantiomers and compounds 2-4 were isolated from this plant for the first time. Compounds 1a, 1b, 2 and 3 increased cell viability in H2O2-treated PC12 cells from (43.41 ± 1.59) % to (61.71 ± 7.56) %, (66.05 ± 5.61) %, (74.34 ± 3.32) % and (69.58 ± 5.02) % at 10 μmol·L-1, respectively.

Effect of Silencing LNK Gene on the Expression of EPO and EPOR in THP-1 Cells / 中国实验血液学杂志

OBJECTIVE@#To investigate the effect of silencing LNK gene on the expression of EPO and EPOR in acute myeloid leukemia cells (THP-1).@*METHODS@#THP-1 cells were cultured. The lentivirus was used as a vector to silence the LNK gene stably. After 72 hours of infection, GFP expression level was detected by the fluorescent inverted microscopy. The lentiviral Infection efficiencies were monitored by flow cytometry. The LNK silencing effect was confirmed. The mRNA expressions of EPO and EPOR were detected by RT-PCR. The protein levels of LNK, EPO and EPOR were detected by Western blot.@*RESULTS@#At the time-point of 72 hours after lentivirus infection, the expression level of GFP was above 85% detected by fluorescent inverted microscopy. The infection efficiency was above 99% by flow cytometry. mRNA expressions of LNK, EPO and EPOR in LNK silencing group were signifycantly lower than those in control group (P＜0.05). The protein levels of LNK, EPO and EPOR in LNK silencing group were significantly lower than those in the control group (P＜0.05).@*CONCLUSION@#THP-1 cell line of LNK gene silencing has been successfully established,the LNK gene has been silenced, the expression of EPO and EPOR decrease, indicating that LNK may participate in the regulation of EPO and EPOR.

EPO and EPOR Expression in Patients with Acute Leukemia and Its Clinical Significance / 中国实验血液学杂志

OBJECTIVE@#To investigate the expression of erythropoietin (EPO) and erythropoietin receptor (EPOR) in patients with acute leukemia (AL) and its clinical significance.@*METHODS@#The levels of EPO and EPOR in plasma were determined by ELISA kit. mRNA expression levels of EPO and EPOR were determined by RT-RCR. The protein expression levels of EPO and EPOR were detected by Western blot.@*RESULTS@#The EPO protein levels in marrow plasma of ALL and AML group were significantly higher than those in the control group (P＜0.05), EPOR protein levels in ALL and AML group were significantly lower than those in the control group (P＜0.05). The mRNA levels of EPO and EPOR in ALL and AML groups were significantly higher than those in the control group (P＜0.05). The mRNA levels of EPO and EPOR in the high risk ALL and AML groups were significantly higher than those in the medium, low risk group and the control group (P＜0.05). The protein expression levels of EPO and EPOR in ALL and AML groups were significantly higher than that in control group (P＜0.05). The mRNA levels of EPO and EPOR in ALL and AML groups did not correlate with hemoglobin level and erythrocyte count (P＞0.05).@*CONCLUSION@#The expressions of EPO and EPOR is higher in ALL and AML patients. The expression levels of EPO and EPOR relate with the risk of ALL and AML. High risk patients have higher expression levels of EPO and EPOR, however, the expression levels of EPO and EPOR do not correlate with hemoglobin level and erythrocyte counting.

Expression and Clinical Significance of STAT3 Genes in Patients with Acute Myeloid Leukemia / 中国实验血液学杂志

OBJECTIVE@#To investigate the expression of STAT3 gene in patients with acute myeloid leukemia and its correlation with clinical characteristics.@*METHODS@#The real-time quantitative RT-PCR was used to detect the level of STAT3 mRNA in bone marrow samples from 38 newly diagnosed patients with acute myeloid leukemia(AML), and its relevance with clinical characteristics and prognosis were statistically analyzed. Western blot was employed to detect the STAT3 protein level in AML patients. The bone marrow cells from 15 healthy subjects were used as control.@*RESULTS@#At the mRNA level, the expression level of STAT3 in the AML group was significantly higher than that in control group (P0.05). The median survival time of patients in STAT3 low expression group was logner than that in high expression group, but the difference was not statistically significant (P>0.005). The level of STAT3 protein in AML patients was significantly higher than that in control group (P<0.05）.@*CONCLUSION@#The STAT3 gene is highly expressed in AML patients, which may be used as a predictor for high-risk of AML.

IRF2-INPP4B-mediated autophagy suppresses apoptosis in acute myeloid leukemia cells

BACKGROUND: The present study aimed to investigate the underlying role of interferon-regulatory factor 2 (IRF2)-inositol polyphosphate-4-phosphatase, type-II (INPP4B) axis in the regulation of autophagy in acute myeloid leukemia (AML) cells. METHODS: Quantitative real time PCR (QRT-PCR) and western blot were performed to determine the expression levels of IRF2, INPP4B and autophagy-related markers in AML cell lines. Autophagy was assessed by elevated Beclin-1 expression, the conversion of light chain 3 (LC3)-I to LC3-II, downregulated p62 expression and green fluorescent protein (GFP)-LC3 puncta formation. The colony formation and apoptosis assays were performed to determine the effects of IRF2 and INPP4B on the growth of AML cells. RESULTS: IRF2 and INPP4B were highly expressed in AML cell lines, and were positively correlated with autophagy-related proteins. Overexpression of IRF2 or INPP4B stimulated autophagy of AML cells, whereas inhibition of IRF2 or INPP4B resulted in the attenuation of autophagy. More importantly, IRF2 or INPP4B overexpression reversed autophagy inhibitor, 3-methyladenine (3-MA)-induced proliferation-inhibitory and pro-apoptotic effects, while IRF2 or INPP4B silencing overturned the proliferation-promoting and anti-apoptotic effects of autophagy activator rapamycin. CONCLUSION: IRF2-INPP4B signaling axis attenuated apoptosis through induction of autophagy in AML cells.

Effect of Silencing and Overexpression of LNK Gene on STAT3 Expression in THP-1 Cells / 中国实验血液学杂志

OBJECTIVE@#To investigate the effect of LNK gene silencing and overexpression on the expression of STAT3 gene in human monocytic leukemia cells (THP-1).@*METHODS@#THP-1 cells were cultured, and the lentivirus was used as a vector to silence and overexpres the LNK gene stably. After transfection for 72 hours, the GFP expression levels were observed by inverted fluorescence microscopy. The lentiviral transfection efficiencies were detected by flow cytometry. The effects of LNK silencing and overexpression were confirmed, and the expression of STAT3 mRNA was detected by RT-PCR. The protein levels of LNK and STAT3 were detected by Western blotting.@*RESULTS@#The GFP expression level of THP-1 cells reached more than 85% after transfection with lentivirus for 72 hours, and the transfection efficiency of cells was above 99%. mRNA expressions levels of LNK and STAT3 in LNK silencing group were signifycantly lower than those in control group, while LNK and STAT3 mRNA levels in the LNK overexpression group was significantly higher than those in control group (P＜0.05). The protein expression levels of LNK and STAT3 in LNK silencing group were significantly lower than those in control group, while that in LNK overexpression group was significantly higher than that in control group (P＜0.05).@*CONCLUSION@#The THP-1 cell line with LNK gene silencing and overexpression has been successfully established. The LNK gene silencing resulted in decrease of STAT3 expression; LNK gene overexpression and leads to inereases of STAT3 expression indicating that LNK participates in the regulation of STAT3.

Variation of STAT3 Gene in Myleproliferative Neoplasms and Its Significance / 中国实验血液学杂志

<p><b>OBJECTIVE</b>To detect the mutation and single nucleotide polymorphisms of STAT3 gene in the patients with myeloproliferative neoplasms (MPN), and to analyze the correlation between STAT3 gene and the subtypes of MPN.</p><p><b>METHODS</b>A total of 147 patients with MPN were selected, including 28 patients with polycythaemia vera (PV), 46 patients with essential thrombocythemia (ET), 10 patients with primary myelofibrosis (PMF), and 63 patients with chronic myeloid leukemia (CML); and 88 healthy persons were used as normal control. DNA of all cases was extracted from bone marrow or peripheral blood, and JAK2V617F gene mutation was detected by allele-specific PCR, then 23 exons of STAT3 gene were amplified by PCR. Mutation and single nucleotide polymorphism of Rs2293152 of STAT3 gene were identified by DNA sequencing.</p><p><b>RESULTS</b>STAT3 gene mutation was found in 8 patients with CML. The mutation rate was 12.7%. the missense mutation（S629T）as found in 3 cases, the synonymous mutation was found in 5 cases (Q469Q 3 cases, G618G 2 cases). One case had mutations at the both sites of S629T and G618G. No mutation of STAT3 gene was found in the normal control group. Rs2293152: detection showed that the G allele of CML group was significantly higher than that of normal control, PV, ET and PMF group (P<0.01), suggesting that the patients with Rs2293152 G allele were more likely to develop CML. The C allele of PV, ET and PMF group was significantly higher than that of CML group (P<0.05), suggesting that the patients with Rs2293152 C allele were more likely to develop PV, ET and PMF. The G allele fiequency of JAK2V617F-negative group was significantly lower than that of the normal control and JAK2V617F positive group (P<0.01), suggesting that the Rs2293152 G allele may be a factor protecting against JAK2V617F mutation.</p><p><b>CONCLUSION</b>In MPN patients, STAT3 gene is unstable and prone to mutation. The different alleles of the Rs2293152 locus of the STAT3 gene are relates with different subtypes and JAK2V617F-negative MPN.</p>

Change of G6PD Activity in Children with Acute Leukemia and Its Clinical Significance / 中国实验血液学杂志

OBJECTIVE@#To explore the change of G6PD activity in children with acute leukemia（AL）and its correlation with the clinical characteristics.@*METHODS@#The G6PD activity in peripheral blood samples from 74 children disagnosed as AL (50 cases of ALL, and 24 cases of AML) was detected by Zinkham method recommended by WHO in 1967, and its relevance with clinical indicators was statistically analyzed. The peripheral blood samples of 70 healthy children were used as the controls.@*RESULTS@#The G6PD activity in ALL and AML groups was significantly lower than that in the control group (P=0.000, P=0.000) and there was no statistical difference between ALL and AML groups. The G6PD activity in bacterial, fungal infection and non-infection groups (no bacterial and fungal infection) were statistically different from control group (P=0.02, P=0.001, P=0.001), respectively. The G6PD activity in bacterial infection group and non-infection group was statistically different from with fungal infection group (P=0.004, P=0.019), respectively. The G6PD activity linearly correlated with leukocyte count and neutrophil percentage in AL children (P=0.000, P=0.001, r=0.465, r=0.434), respectively. The median survival time of G6PD activity deficiency group was higher than that in the normal group, but without statistically significant difference (P=0.4149).@*CONCLUSION@#The G6PD activity in AL children is significantly lower than that in healthy children, and the G6PD activity linearly relates with leukocyte count and neutrophil percentage of AL children. The patients with G6PD activity deficiency is more susceptible to fungal infection, moreover the infection is more serious.

Research progress on quality evaluation and assay methods of polysorbate 80 / 药物评价研究

Research progress ofpolysorbate 80 was summarized,including standards,chemical composition and impurity test.Assay methods of polysorbate 80 in preparations were also summarized.In addition,characteristics of the current technologies were discussed.Compositions of polysorbate 80 could be detected directly by spectrophotometry,SEC-ELSD or LC-MS methods.In addition,they could be analyzed indirectly by determing the hydrolysates with HPLC-UV or GC methods.In recent years,remarkable progress has been achieved in chemical composition determination and impurity test ofpolysorbate 80.But assay method of the excipient in related drugs need further study,due to large difference between purities ofpolysorbate 80 raw materials.

Toxic effect of Niuhuang Zhenjing Pill on zebrafish embryo development / 药物评价研究

Objective To study the toxic effect ofNiuhuang Zhenjing Pill on zebrafish embryo development.Methods Zebrafish embryo was used to investigate the development situation under stereo microscope at different drug concentration (40,200,400,800,1 200 tg/mL) including embryo teratogenic and lethal tests.Results No abnormal embryo development was found when the concentration was not greater than 200 μg/mL (Approximately equal to 30 times the normal adult dosage,exceeding the normal effective dose).While,there was toxicity to embryo development at the concentration greater than 200 tg/mL.The head and eyes of the embryo were smaller,and the pupils were colorless.Zebrafish embryonic heart malformation was observed,heart rate was weak,blood pool congestion happened and back blood flow was not smooth.The abdominal surface of the embryo was poor in transparency,the tail was twisted,the yolk sac elongation structure became thicker,the notochord became thin,there was no normal embryo movement,and the body length became shorter.After the withdrawal,the abnormal phenotype was not recovered,and severity of the abnormal phenotype was concentration-dependent.It was also observed that all embryos died within 1 d after fertilization at the concentration of 1 200 μg/mL.Conclusion The drug could influence the development of zebrafish embryos when the concentration was higher than 200 tg/mL.Its toxic effect occurs during embryonic organogenesis.And it mainly affects the development of embryonic nervous system,cardiovascular system and bone.

Connotation of "precision novelty assessment of literature" and its empirical analysis / 中华医学图书情报杂志

After the concept of"precision novelty assessment of literature" was proposed by integrating"precision"into novelty assessment of literature, the connotation of"precision novelty assessment of literature" was elaborated, including full communication with the trustee, working out comprehensive strategies for "precision novelty assess-ment of literature", writing accurate and complete retrieval process of"precision novelty assessment of literature", appropriate and objective report of "precision novelty assessment of literature". Empirical analysis was then con-ducted by elaborating the concept of"precision novelty assessment of literature" in order to enrich the theory for the novelty assessment of scientific literature and further improve its practical level.

Research progress on quality evaluation and assay methods of polysorbate 80 / 药物评价研究

Research progress ofpolysorbate 80 was summarized,including standards,chemical composition and impurity test.Assay methods of polysorbate 80 in preparations were also summarized.In addition,characteristics of the current technologies were discussed.Compositions of polysorbate 80 could be detected directly by spectrophotometry,SEC-ELSD or LC-MS methods.In addition,they could be analyzed indirectly by determing the hydrolysates with HPLC-UV or GC methods.In recent years,remarkable progress has been achieved in chemical composition determination and impurity test ofpolysorbate 80.But assay method of the excipient in related drugs need further study,due to large difference between purities ofpolysorbate 80 raw materials.

Toxic effect of Niuhuang Zhenjing Pill on zebrafish embryo development / 药物评价研究

Objective To study the toxic effect ofNiuhuang Zhenjing Pill on zebrafish embryo development.Methods Zebrafish embryo was used to investigate the development situation under stereo microscope at different drug concentration (40,200,400,800,1 200 tg/mL) including embryo teratogenic and lethal tests.Results No abnormal embryo development was found when the concentration was not greater than 200 μg/mL (Approximately equal to 30 times the normal adult dosage,exceeding the normal effective dose).While,there was toxicity to embryo development at the concentration greater than 200 tg/mL.The head and eyes of the embryo were smaller,and the pupils were colorless.Zebrafish embryonic heart malformation was observed,heart rate was weak,blood pool congestion happened and back blood flow was not smooth.The abdominal surface of the embryo was poor in transparency,the tail was twisted,the yolk sac elongation structure became thicker,the notochord became thin,there was no normal embryo movement,and the body length became shorter.After the withdrawal,the abnormal phenotype was not recovered,and severity of the abnormal phenotype was concentration-dependent.It was also observed that all embryos died within 1 d after fertilization at the concentration of 1 200 μg/mL.Conclusion The drug could influence the development of zebrafish embryos when the concentration was higher than 200 tg/mL.Its toxic effect occurs during embryonic organogenesis.And it mainly affects the development of embryonic nervous system,cardiovascular system and bone.

Connotation of "precision novelty assessment of literature" and its empirical analysis / 中华医学图书情报杂志

After the concept of"precision novelty assessment of literature" was proposed by integrating"precision"into novelty assessment of literature, the connotation of"precision novelty assessment of literature" was elaborated, including full communication with the trustee, working out comprehensive strategies for "precision novelty assess-ment of literature", writing accurate and complete retrieval process of"precision novelty assessment of literature", appropriate and objective report of "precision novelty assessment of literature". Empirical analysis was then con-ducted by elaborating the concept of"precision novelty assessment of literature" in order to enrich the theory for the novelty assessment of scientific literature and further improve its practical level.

Isoflavones and flavans from Millettia nitida var. hirsutissima / 中国中药杂志

The current study to separate and identify constituents from Millettia nitida var. hirsutissima. The compounds from Millettia nitida var. hirsutissima were isolated by means of various chromatographic techniques such as column chromatography over ODS and Sephadex LH-20, preparative HPLC, and the structures of these isolated compounds were identified through spectroscopic analyses. Nine isoflavonoids and two flavans were isolated and identified as 5-O-methy genistein (1), 7-hydroxy-3',4'-dimethoxyisoflavone (2), ononin (3), catechin (4), formononetin (5), genistein (6), calycosin (7), (-)-gallocatechin (8), sissotrin (9), wistin (10), daidzin (11). Compounds 1, 2, 9 are obtained from the genus Millettia for the first time,and compounds 4, 8 are isolated from this plant for the first time.

A260G and A386G single nucleotide polymorphisms of the DAZL gene are not correlated with male infertility in the Chinese population of Zhejiang Province / 中华男科学杂志

<p><b>OBJECTIVE</b>To assess the association of the A260G and A386G single nucleotide polymorphisms (SNP) of the DAZL gene with male infertility in the Chinese population of Zhejiang Province.</p><p><b>METHODS</b>We collected the peripheral blood samples from 317 idiopathic infertile males with azoospermia or oligozoospermia and 246 normal fertile men, and genotyped the polymorphic loci of the A260G and A386G polymorphisms of the DAZL gene using the SNaPshot technique.</p><p><b>RESULTS</b>The DAZL gene A260G was found genetically polymorphic in the Chinese population of Zhejiang Province, with the gene frequencies and their distribution consistent to the Hardy-Weinberg equilibrium. The frequencies of the AA, AG and GG genotypes of the A260G polymorphism were 92.3%, 7.3%, and 0.4% respectively in the normal controls and 94.3%, 5.7%, and 0% in the infertile patients, with no statistically significant differences between the two groups (P = 0.43, OR = 0.78, 95% CI 0.413-1.46). Heterozygosis (AG) of A386G was found in 1 of the control males but not in the infertile patients, while homozygosis (GG) of A386G was not observed in either group (P = 0.259, OR = 0.698, 59% CI: 0.374-1.306).</p><p><b>CONCLUSION</b>A260G and A386G SNPs of the DAZL gene are not associated with spermatogenic failure and neither represents a molecular marker for the genetic diagnosis of male infertility in the Chinese population of Zhejiang Province.</p>