Chinese Medicine Regulates DNA Methylation to Treat Haematological Malignancies: A New Paradigm of "State-Target Medicine".

Aberrant regulation of DNA methylation plays a crucial causative role in haematological malignancies (HMs). Targeted therapy, aiming for DNA methylation, is an effective mainstay of modern medicine; however, many issues remain to be addressed. The progress of epigenetic studies and the proposed theory of "state-target medicine" have provided conditions to form a new treatment paradigm that combines the "body state adjustment" of CM with targeted therapy. We discussed the correlation between Chinese medicine (CM) syndromes/states and DNA methylation in this paper. Additionally, the latest research findings on the intervention and regulation of DNA methylation in HMs, including the core targets, therapy status, CM compounds and active components of the Chinese materia medica were concisely summarized to establish a theoretical foundation of "state-target synchronous conditioning" pattern of integrative medicine for HMs, simultaneously leading a new perspective in clinical diagnosis and therapy.

Chinese Medicine Regulates DNA Methylation to Treat Haematological Malignancies: A New Paradigm of "State-Target Medicine" / 中国结合医学杂志

Aberrant regulation of DNA methylation plays a crucial causative role in haematological malignancies (HMs). Targeted therapy, aiming for DNA methylation, is an effective mainstay of modern medicine; however, many issues remain to be addressed. The progress of epigenetic studies and the proposed theory of "state-target medicine" have provided conditions to form a new treatment paradigm that combines the "body state adjustment" of CM with targeted therapy. We discussed the correlation between Chinese medicine (CM) syndromes/states and DNA methylation in this paper. Additionally, the latest research findings on the intervention and regulation of DNA methylation in HMs, including the core targets, therapy status, CM compounds and active components of the Chinese materia medica were concisely summarized to establish a theoretical foundation of "state-target synchronous conditioning" pattern of integrative medicine for HMs, simultaneously leading a new perspective in clinical diagnosis and therapy.

Effect of Petroleum Ether Extract of Rhizoma Amorphophalli on Biological Characteristics of K562 Cells and Its Mechanism / 中国实验血液学杂志

OBJECTIVE@#To investigate the role of petroleum ether extract of Rhizoma Amorphophalli (SLG) in inhibiting proliferation and promoting apoptosis and differentiation of leukemia K562 cells.@*METHODS@#K562 cells were processed by SLG and PD98059 which was the ERK signaling pathway blocker. Then cell vitality was tested by MTT. Cell apoptosis rate and positive percentage of antigen expression related with differentiation were detected by flow cytometry. The protein expression levels of ERK1/2 and pERK1/2 were detected by Western blot.@*RESULTS@#The proliferation activity of K562 was reduced by 50, 100, 200 mg/L SLG in a concentration dependent manner (r=0.9997). The apoptosis rate and positive expression rate of CD11b, CD14 and CD42b which were related with differentiation were raised by SLG, as well as the expression of pERK1/2, while PD98059 could reverse the promoting effect of SLG on apoptosis and differentiation partially.@*CONCLUSION@#SLG can inhibit the proliferation and promote apoptosis and differentiation of K562 cells through ERK signaling pathway.

Chinese Medicine Treatment on Graft-Versus-Host Disease after Allogeneic Hematopoietic Stem Cell Transplantation / 中国结合医学杂志

Graft-versus-host disease (GVHD) is the most common complication after allogeneic hematopoietic stem cell transplantation, and also an important factor affecting the survival and quality of life in patients after transplantation. Currently, immunosuppressive therapy is commonly used for GVHD, but the curative effect is not ideal. How to effectively prevent and treat GVHD is one of the difficulties to be solved urgently in the field of transplantation. In this paper, we summarize the latest progress in pathogenesis, prevention and treatment of GVHD with Chinese medicine (CM). We hope it will provide ideas and methods for exploring the mechanism and establishing a new comprehensive therapy for GVHD with CM.

Effects of attention to positive and negative and rumination on post traumatic growth in patients with coronary artery disease undergoing interventional therapy / 中国实用护理杂志

Objective To investigate the level of posttraumatic growth(PTG) in patients with percutaneous coronary intervention(PCI), to explore the ruminant, positive/negative information attention to the impact of both on the posttraumatic growth. Methods A study of 360 patients with coronary artery disease was conducted using general information, Event Related Rumination Inventory (ERRI) and Attention to Positive and Negative Information Scale (APNIS) and the Post Traumatic Growth Inventory (PTGI). Results The Attention to Positive information (API) score was (38.47 ± 9.33) points, and the Attention to Negative Information (ANI) score was (32.99 ± 8.48) points for interventional patients with coronary heart disease, the total score of PTGI was(49.96±19.96)points and the total score of ERRI was (26.52 ± 10.55)points. In PCI patient the PTG and API were positively correlated with rumination (both purposeful and invasive), and PTG was negatively correlated with ANI. The result showed that API, rumination (including deliberate and intrusive) had positive predictive effect on PTG, and ANI had negative predictive effect on PTG, API and ANI respectively had significant interaction with ruminant meditation. Conclusions Patients with coronary artery disease undergoing interventional therapy reported the API was low, the ANI level was higher, the PTG and rumination level was lower. Clinical measures higher, the PTG and rumination level lower. Clinical measures should be taken to guide patients to pay more attention to positive information, to reduce the concern for negative information, and to guide patients to correct reflection in order to promote post-traumatic growth.

Ginseng-Derived Panaxadiol Saponins Promote Hematopoiesis Recovery in Cyclophosphamide-Induced Myelosuppressive Mice: Potential Novel Treatment of Chemotherapy-Induced Cytopenias / 中国结合医学杂志

<p><b>OBJECTIVE</b>To investigate the potential efficacy of panaxadiol saponins component (PDS-C), a biologically active fraction isolated from total ginsenosides, to reverse chemotherapy-induced myelosuppression and pancytopenia caused by cyclophamide (CTX).</p><p><b>METHODS</b>Mice with myelosuppression induced by CTX were treated with PDS-C at a low- (20 mg/kg), moderate- (40 mg/kg), or high-dose (80 mg/kg) for 7 consecutive days. The level of peripheral white blood cell (WBC), neutrophil (NEU) and platelet (PLT) were measured, the histopathology and colony formation were observed, the protein kinase and transcription factors in hematopoietic cells were determined by immunohistochemical staining and Western blot.</p><p><b>RESULTS</b>In response to PDS-C therapy, the peripheral WBC, NEU and PLT counts of CTX-induced myelosuppressed mice were significantly increased in a dose-dependent manner. Similarly, bone marrow histopathology examination showed reversal of CTX-induced myelosuppression with increase in overall bone marrow cellularity and the number of hematopoietic cells (P<0.01). PDS-C also promoted proliferation of granulocytic and megakaryocyte progenitor cells in CTX-treated mice, as evidenced by significantly increase in colony formation units-granulocytes/monocytes and -megakaryocytes (P<0.01). The enhancement of hematopoiesis by PDS-C appears to be mediated by an intracellular signaling pathway, this was evidenced by the up-regulation of phosphorylated mitogen-activated protein kinase (p-MEK) and extracellular signal-regulated kinases (p-ERK), and receptor tyrosine kinase (C-kit) and globin transcription factor 1 (GATA-1) in hematopoietic cells of CTX-treated mice (P<0.05).</p><p><b>CONCLUSIONS</b>PDS-C possesses hematopoietic growth factor-like activities that promote proliferation and also possibly differentiation of hematopoietic progenitor cells in myelosuppressed mice, probably mediated by a mechanism involving MEK and ERK protein kinases, and C-kit and GATA-1 transcription factors. PDS-C may potentially be a novel treatment of myelosuppression and pancytopenia caused by chemotherapy.</p>

Clinical study of pai-neng-da capsule in the treatment of chronic aplastic anemia / 中国结合医学杂志

<p><b>OBJECTIVE</b>To evaluate the efficacy and safety of Pai-Neng-Da Capsule (panaxadiol saponins component, PND), a new Chinese patent medicine, on patients with chronic aplastic anemia (CAA) and to explore the optimal therapeutic regimen for CAA.</p><p><b>METHOD</b>A total of 36 patients with CAA were enrolled and divided into three groups: the AP group (20 cases, andriol 120 mg/day + PND 240 mg/day), the ACP group (13 cases, andriol 120 mg/day + cyclosporine 3-6 mg kd(-1) day(-1) + PND 240 mg/day), and the PND group (3 cases, PND 240 mg/day). All patients were treated and followed up for 6 months. Peripheral blood counts, renal and hepatic function and Chinese medical (CM) symptoms of patients were assessed and all indices were gathered at the beginning and end of the study.</p><p><b>RESULT</b>In the AP group, no significant hematologic difference was observed at the end of 6-month treatment comparing with the beginning. In the ACP group, the blood counts were maintained at the same level after the 6-month treatment. In the PND group, trilineage hematologic improvement was displayed at the end of 6-month treatment comparing with the beginning. No significant difference was showed in renal and hepatic function in all patients. All patients' clinical symptom improved according to CM symptom score. The effective rates were 95%, 73% and 100%, respectively.</p><p><b>CONCLUSION</b>PND improved the efficacy and decreased side effects by cutting down the dosage of andriol, and it could also improve patients' clinical symptom and quality of life. PND were effective and safe in the treatment of CAA, it could be used alone or in combination with pharmacological agents such as andriol and cyclosporine.</p>

Effects of panaxadiol saponins component as a new Chinese patent medicine on proliferation, differentiation and corresponding gene expression profile of megakaryocytes / 中国结合医学杂志

<p><b>OBJECTIVE</b>To investigate the effects of panaxadiol saponins component (PDS-C) isolated from total saponins of panax ginseng on proliferation, differentiation and corresponding gene expression profile of megakaryocytes.</p><p><b>METHODS</b>Bone marrow culture of colony forming assay of megakaryocytic progenitor cells (CFU-MK) was observed for the promoting proliferation mediated by PDS-C, and differentiation of megakaryocytic blasts caused by PDS-C was analyzed with flow cytometry in CHRF-288 and Meg-01 cells, as well as proliferation, differentiation-related genes expression profile and protein expression levels were detected by human gene expression microarray and western blot.</p><p><b>RESULTS</b>In response to PDS-C 10, 20 and 50 mg/L, CFU-MK from 10 human bone marrow samples was increased by 28.9%±2.7%, 41.0%±3.2% and 40.5%±2.6% over untreated control, respectively (P <0.01, each). Flow cytometry analysis showed that PDS-C treated CHRF-288 cells and Meg-01 cells significantly increased in CD42b, CD41, TSP and CD36 positive ratio, respectively. PDS-C induced 29 genes up-regulated more than two-fold commonly in both cells detected by human expression microarray representing 4000 known genes. The protein expression levels of ZNF91, c-Fos, BTF3a, GATA-1, RGS2, NDRG2 and RUNX1 were increased with western blot in correspond to microarray results.</p><p><b>CONCLUSION</b>PDS-C as an effective component for hematopoiesis, play the role to enhance proliferation and differentiation of megakaryocytes, also up-regulated expression of proliferation, differentiation-related genes and proteins in vitro.</p>

Effects of Danshen Injection () on inhibiting proliferation and inducing apoptosis through down-regulation of mutant JAK2 gene and its protein phosphorylation in human erythroid leukemic cells / 中国结合医学杂志

<p><b>OBJECTIVE</b>To explore the effects of Danshen Injection () on inhibition proliferation, inducing apoptosis and its possible mechanisms on human erythroid leukemic (HEL) cells.</p><p><b>METHODS</b>The commercial Chinese patent medicine of Danshen Injection was extracted and isolated from Chinese herb of Salvia miltiorrhiza bung. The inhibition effects of proliferation were assayed by 3-(4,5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide (MTT) method in HEL cells treated by Danshen Injection at various concentrations for 48 h. The cellular apoptosis was observed in morphology, analyzed by flow cytometry with annexin V and propidium iodide (PI) staining, and examined by DNA degradation ladder on agarose gel electrophoresis. Meanwhile, the expression levels of mutant Janus kinasez (JAK2) gene and phosphorylation-JAK2 (P-JAK2) protein were detected by allele specific-polymerase chain reaction and Western blot.</p><p><b>RESULTS</b>The proliferation of HEL cells was effectively inhibited by Danshen Injection in a dose-dependent manner, with suppression rates from 19.46±2.31% to 50.20±5.21%. Typical apoptosis cells was observed in Danshen Injection treated HEL cells, the rates of annexin V positive cells increased obviously in a dose-dependent manner, as well as the DNA degradation ladder of apoptosis revealed on gel electrophoresis. The expression levels of mutant JAK2 gene and P-JAK2 protein reduced gradually with increasing dosage of Danshen injection.</p><p><b>CONCLUSION</b>Danshen Injection could not only significantly inhibit the proliferation, but also induce apoptosis in HEL cells; down-regulation of the mutant JAK2 gene and P-JAK2 protein expressions are probably one of its molecular mechanisms.</p>

Rubus parvifolius L. inhibited the growth of leukemia K562 cells in vitro and in vivo / 中国结合医学杂志

<p><b>OBJECTIVE</b>To determine the antiproliferative activity of Rubus parvifolius L. (RP) extract, its medicinal serum and RP total saponins (RPTS) against K562 cells in vitro and in vivo.</p><p><b>METHODS</b>Nude mice models bearing leukemia tumors were treated with different concentrations of RP extract. The size, weight and histopathological change of leukemic tumors were determined. Semi-solid agar culture and methylthiazolyl tetrazolium (MTT) assay were used to determine in vitro the inhibition of colony formation and proliferation of K562 cells respectively by different concentrations of RP medicinal serum and RPTS.</p><p><b>RESULTS</b>RP extract had a tumor inhibition rate of 84.8% when administered to mice at a dose of 1.0 g/day of crude RP root equivalent. Semi-solid agar culture of K562 cells in the presence of 20% (v/v) of RP medicinal serum and 150 mg/L RPTS demonstrated a 50.8% and 100% inhibition of the colony forming unit (CFU)-K562, respectively. The same doses of RP medicinal serum and RPTS showed a proliferation inhibition of 31.4% and 86.3%, respectively against K562 cells in MTT assay.</p><p><b>CONCLUSION</b>RP extract and RPTS show effective antiproliferative activity against myeloid leukemia cells in vitro and in vivo.</p>

Outcomes of patients with combined methylmalonic acidemia and homocystinuria after treatment / 中华儿科杂志

<p><b>OBJECTIVE</b>Combined methylmalonic acidemia with homocystinuria is a common form of methylmalonic acidemia in China. Patients with this disease can progress to death without timely and effective treatment. This study aimed to analyze the treatment outcomes of patients with combined methylmalonic acidemia and homocystinuria.</p><p><b>METHOD</b>From September 2004 to April 2012, 58 patients with combined methylmalonic acidemia and homocystinuria (34 males and 24 females) were diagnosed and treated in our hospital. Fifty cases were from clinical patients including 42 early-onset cases and 8 late-onset cases. Their age when they were diagnosed ranged from 18 days to 30.8 years. The other 8 cases were from newborn screening. All the patients were treated with vitamin B12, betaine, folic acid, vitamin B6, and L-carnitine. The physical and neuropsychological development, general laboratory tests, the levels of amino acids, acylcarnitines, and homocysteine in blood, and organic acids in urine were followed up.</p><p><b>RESULT</b>The follow-up period ranged from 1 month to 7.1 years. Three cases died (all were early-onset cases). In the other patients after treatment, the symptoms such as recurrent vomiting, seizures, lethargy, and poor feeding disappeared, muscle strength and muscle tension were improved, and general biochemical abnormalities such as anemia and metabolic acidosis were corrected. Among the surviving 55 cases, 49 had neurological impairments such as developmental delay and mental retardation. The median levels of blood propionylcarnitine and its ratio with acetylcarnitine, serum homocysteine, and urine methylmalonic acid were significantly decreased (P < 0.01), from 7.73 µmol/L (ranged from 1.5 to 18.61 µmol/L), 0.74 (ranged from 0.29 to 2.06), 97.3 µmol/L (ranged from 25.1 to 250 µmol/L) and 168.55 (ranged from 3.66 to 1032.82) before treatment to 2.74 µmol/L (ranged from 0.47 to 12.09 µmol/L), 0.16 (ranged from 0.03 to 0.62), 43.8 µmol/L (ranged from 17 to 97.8 µmol/L) and 6.81 (ranged from 0 to 95.43) after treatment, respectively.</p><p><b>CONCLUSION</b>Patients with combined methylmalonic acidemia and homocystinuria respond to a combined treatment consisting of supplementation of hydroxycobalamin, betaine, folic acid, vitamin B6 and L-carnitine with clinical and biochemical improvement. But the long-term outcomes are unsatisfactory, with neurological sequelae in most patients.</p>

Effect of Kansui Radix prepared by different processes on LO2 cell cycle and apoptosis / 中国中药杂志

<p><b>OBJECTIVE</b>To discuss the effect of Kansui Radix prepared by different processes on cell cycle and apoptosis of normal human liver cell lines LO2.</p><p><b>METHOD</b>With normal human liver cell lines LO2 as the study object, the MTT method was adopted to study the effect of Kansui Radix prepared by different processes, including Kansui Radix, stir-baking Kansui Radix, Kansui Radix moistening with vinegar and Kansui prepared by different processes, on LO2 cell activity. The cellular morphological changes were observed by inverted microscope. The effect of Kansui Radix stir-baked with vinegar on LO2 cell cycle and apoptosis was observed by flow cytometry.</p><p><b>RESULT</b>Compared with the negative control group, Kansui could obviously inhibit the activity of human normal liver cell lines LO2 (P <0.01) , and significantly increase the percentage of LO2 cells in S phase (P <0.05) , notably decrease the percentage of LO2 cells in G2/M phase (P <0.01) , significantly increase the early apoptosis rate, late apoptosis rate and necrosis rate and total apoptosis rate of human normal liver cell lines LO2 (P <0.01). Compared with the Kansui group, all of the other processed Kansui samples could significantly decrease the cell proliferation inhibition (P <0.01) , and the trend of morphological degradation. Besides, they could significantly increase the percentage of LO2 cells in G2/M phase (P <0.05, P <0.05, P <0. 01) , significantly decrease the early apoptosis rate, late apoptosis rate and necrosis rate, and total apoptosis rate of human normal liver cell lines LO2 (P < 0.01). The order of the increase in the percentage of cells in G2/M phase and the decrease in apoptosis rate was Kansui Radix stirbaked with vinegar > Kansui Radix moistening with vinegar > stir-baking Kansui Radix.</p><p><b>CONCLUSION</b>The toxicity of processed Kansui could be reduced by affecting LO2 cell cycle and apoptosis. The processes of stir-baking and moistening with vinegar can play a synergistic effect in the detoxication of human normal liver cell lines LO2, which provides a basis for unveiling the rationality of stirbaking with vinegar of Kansui in the detoxication, as well as the optimizing the process.</p>

Diagnosis and treatment of isolated methylmalonic acidemia / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To explore the clinical feature, therapeutic effect and prognosis of isolated methylmalonic acidemia.</p><p><b>METHODS</b>The clinical characteristics, laboratory findings, treatment and outcome of 40 patients were retrospectively analyzed. The main treatment was a low-protein diet supplemented with L-carnitine and special milk free of leucine, valine, threonine and methionine. Vitamin B12 was also given to cobalamin responders. The patients were followed up every 1-3 months.</p><p><b>RESULTS</b>Mutations in the MUT gene were identified in 30 of 33 patients who had accepted DNA testing. Thirty cases were treated and followed up regularly for from 1 month to 8 years. Eight cases had died, 8 had developed normal intelligence, among whom 4 from newborn screening were asymptomatic. Psychomotor developmental delay and mental retardation were present in 14 cases. The propionylcarnitine level, ratio of propionylcarnitine/acetylcarnitine in blood, methylmalonic acid and methylcitric acid levels in urine have decreased significantly, with the median values reduced respectively from 24.15 (7.92-81.02) μmol/L, 1.08 (0.38-6.01), 705.34 (113.79-3078.60) and 7.71 (0.52-128.21) to 10.50 (3.00-30.92) μmol/L, 0.63 (0.25-2.89), 166.23 (22.40-3322.21) and 3.96 (0.94-119.13) (P < 0.05).</p><p><b>CONCLUSION</b>The prognosis of isolated methylmalonic acidemia may be predicted with the enzymatic subgroup, age at onset and cobalamin responsiveness. Outcome is unfavorable in neonatal patients and those who were non-responsive to cobalamin.</p>

Primary carnitine deficiency in 17 patients: diagnosis, treatment and follow up / 中华儿科杂志

<p><b>OBJECTIVE</b>Many children were found to have low free carnitine level in blood by tandem mass spectrometry technology. In some of the cases the problems occurred secondary to malnutrition, organic acidemia and other fatty acid oxidation metabolic diseases, and some of cases had primary carnitine deficiency (PCD). In the present article, we discuss the diagnosis of PCD and evaluate the efficacy of carnitine in the treatment of PCD.</p><p><b>METHOD</b>We measured the free carnitine (C0) and acylcarnitine levels in the blood of 270 000 neonates from newborns screening program and 12 000 children with suspected clinical inherited metabolic diseases by tandem mass spectrometry. The mutations of carnitine transporter protein were tested to the children with low C0 level and the diagnosis was made. The children with PCD were treated with 100 - 300 mg/kg of carnitine.</p><p><b>RESULT</b>Seventeen children were diagnosed with PCD, 6 from newborn screening program and 11 from clinical patients. Mutations were found in all of them. The average C0 level [(2.9 ± 2.0) µmol/L] in patients was lower than the reference value (10 µmol/L), along with decreased level of different acylcarnitines. The clinical manifestations were diverse. For the 6 patients from newborn screening, 4 were asymptomatic, 1 showed hypoglycaemia and 1 showed movement intolerance from 2 years of age. For the 11 clinical patients, 8 showed hepatomegaly, 7 showed myasthenia, 6 showed cardiomyopathy, 1 showed chronic abdominal pain, and 1 showed restlessness and learning difficulty. Among these patients, 14 cases were treated with carnitine. Their clinical symptoms disappeared 1 to 3 months later. The C0 level in the blood rose to normal, with the average from (4.0 ± 2.7) µmol/L to (20.6 ± 8.3) µmol/L (P < 0.01). However, the level was still lower than the average level of healthy children [(27.1 ± 4.5) µmol/L, P < 0.01].</p><p><b>CONCLUSION</b>Seventeen patients were diagnosed with PCD by the test levels of free carnitine and acylcarnitines in blood with tandem mass spectrometry, and gene mutation test. Large dose of carnitine had a good effect in treatment of the PCD patients.</p>

Research and development of the effective components of panaxdiol saponin as new Chinese patent medicine for treating hemocytopenia / 中国结合医学杂志

Pancytopenia (hemocytopenia) such as pr imary immune primary thrombocytopenia (ITP), aplastic anemia and chronic neutropenia (agnogenic leukocytopenia) were of ten t reated by glucocor t icoids, androgen and often treated glucocorticoids, immunosuppressive agents at present, but the response to these treatments has not been always satisfactory, and may cause serious adverse events. Our research has identified a biological active component in ginseng extract and the active component, panaxadiol saponins component (PDS-C), was isolated from total saponins of ginsenosides, and formulated into capsules named as Painengda. We successfully obtained approval from State Food and Drug Administration (SFDA) of China in 2010 to conduct clinical trials of PDS-C as class-five new Chinese patent medicine. Phase I and phase II clinical trials of PDS-C and Painengda Capsule were carried out in the treatment of ITP and agnogenic leukocytopenia. The composition and content of PDS-C have been analyzed and defined by high-performance liquid chromatography-chromatographymass spectrometry (HPLC-MS) and HPLC using specific monomers of ginsenosides as the reference standards. mass PDS-C is very efficacious for treating mice and rats with ITP and aplastic anemia, and myelosuppression caused by chemotherapy or radiation. Our animal model studies and cell biology and molecular biology experiments demonstrated that PDS-C possessed dual activities, namely that of promoting proliferation and differentiation of hematopoietic progenitor cells, and that of regulating the immune function. PDS-C and Painengda Capsule as a new Chinese patent medicine have been successfully transferred to industry. We believe that PDS-C is effective and safe in the treatment of refractory hemocytopenia. The advantages are that it is effective in small doses, it is convenient to use because of its oral administration, its lack of adverse events, it could be used alone or in combination with pharmacological agents, which improve the efficacy and decrease adverse events.

Effects of epimedium total flavonoids on bone mineral density and bone metabolism-related indices in primary osteoporosis / 中国组织工程研究

BACKGROUND: Previous animal experiments have shown that epimedium total flavonoids can exhibit preventive effects on estrogen-related bone loss, but few data are available in clinical research.OBJECTIVE: To investigate the effects of pimedium total flavonoids on bone mineral density (BMD) inprimary osteoporosis.DESIGN, TIME AND SETTING: A randomized, double-blinded, positively controlled clinical observation was performed at the Department of Traditional Chinese Medicine, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology between June 2005 and September 2007.PARTICIPANTS: A total of 64 patients with primary osteoporosis consisting of 11 males and 53 females were included in this study. METHODS: All patients were randomly and evenly assigned into a treatment group and a control group. The treatment group was orally administered epimedium total flavonoids, 0.45 g once, three times daily, for a total of 6 months. Simultaneously, the control group was orally administered Gushukang, 10 g once, twice daily, for a total of 6 months. MAIN OUTCOME MEASURES: BMD in the sites of lumbar vertebrae (L1-4), neck of femur, Wards triangle, greater trochanter, and left hip was measured, and simultaneously serum levels of calcium, phosphorus, and alkaline phosphatase (ALP) were detected pdor to and after treatment in each group.RESULTS: All 64 patients with primary osteoporosis were included In the final analysis, The BMD in the lumbar vertebrae (L1-4) was significantly higher in the treatment group than in the control group (P<0.05) and there was no significant difference In BMD in other sites between the two groups (P>0.05). Compared with pdor to drug application, BMD in the treatment group did not present obvious change after epimedium total flavonoids application, while in the control group, BMD in the sites of lumbar vertebrae (L1-4), neck of femur, Wards triangle, and greater trochanter was significantly decreased (P<0.01-0.05). After drug application, serum level of calcium was significantly increased in each group, compared with prior to drug application (P<0.05), and no significant difference existed between the treatment and control groups (P>0.05). Tn the treatment group, serum levels of phosphorus and ALP did not alter significantly compared with prior to epimedium total flavonoids application (P>0.05).CONCLUSION: Epimedium total flavonoids exhibit effective effects on the maintenance of BMD in primary osteoporosis.

Cortical neuron apoptosis induced by beta-amyloid peptide and protective effect of panoxadiol in mice / 浙江大学学报·医学版

<p><b>OBJECTIVE</b>To investigate the apoptosis of cortical neurons induced by beta-amyloid peptide (Abeta(1-40)) and the protective effect of panoxadiol.</p><p><b>METHODS</b>The Abeta(1-40) induced damage of primarily cultured mouse cortical neurons was examined with morphological observation, MTT assay, DNA agarose gel electrophoresis and Western-blot.</p><p><b>RESULT</b>After 48 h treated with 12 mumol/L Abeta(1-40), the cortical neurons showed apoptotic characteristics: including decreased OD570 value in MTT assay, DNA cleavage fragment in electrophoresis and increased apoptotic cells. Western-blot showed that the expression of bcl-2 reduced significantly (P<0.05). Cell apoptosis was significantly attenuated in 40 mg/L panoxadiol treated group.</p><p><b>CONCLUSION</b>Panoxadiol can protect cultured cortical neurons from apoptosis induced by Abeta(1-40) in mice.</p>

Apoptosis of HL-60 cells induced by aescinate / 中国实验血液学杂志

The aim of this study was to investigate the effects of aescinate on inhibition and apoptosis of HL-60 cell line from promyelocytic leukemia. HL-60 cells at logarithm phase were treated with aescinate. Cell survival rate and cell morphology were observed, and the cell apoptosis was analyzed by Annexin V/PI-FITC double labeling and DNA electrophoresis. The results showed that HL-60 cells could be inhibited in the presence of 15-120 mg/L of aescinate for 48 hours, survival rates were (92.2+/-0.69)%-(8.2+/-0.96)%, which were significantly lower than that of non-aescinate control (99.4+/-0.31)% (all p<0.01). The apoptosis of cells could be induced by aescinate treatment at dosage of 15-60 mg/L for 24 hours, the Annexin V positive cells accounted for (12.7+/-0.58)%-(65.4+/-1.30)% which were significantly higher than that of non-aescinate control (0.57+/-0.03)% (all p<0.01). The typical DNA ladder of HL-60 cells treated with aescinate was shown on the DNA electrophoresis pattern. It is concluded that aescinate can specifically induce apoptosis of leukemic HL-60 cells, which provides an experimental evidence for treatment of leukemia with aescinate as a supplementary agent to chemotherapy.

Clinical observation on trigonella foenum-graecum L. total saponins in combination with sulfonylureas in the treatment of type 2 diabetes mellitus / 中国结合医学杂志

<p><b>OBJECTIVE</b>To evaluate the efficacy and safety of trigonella foenum-graecum L. total saponins (TFGs) in combination with sulfonylureas (SU) in the treatment of patients with type 2 diabetes mellitus (T2DM) not well controlled by SU alone.</p><p><b>METHODS</b>Sixty-nine T2DM patients whose blood glucose levels were not well controlled by oral sulfonylureas hypoglycemic drug were randomly assigned to the treated group (46 cases) and the control group (23 cases), and were given TFGs or placebo three times per day, 6 pills each time for 12 weeks, respectively. Meanwhile, the patients continued taking their original hypoglycemic drugs. The following indexes, including effects on traditional Chinese medicine (TCM) symptoms, fast blood glucose (FBG), 2-h post-prandial blood glucose (2h PBG), glycosylated hemoglobin (HbA1c), clinical symptomatic quantitative scores (CSQS), body mass index (BMI), as well as hepatic and renal functions, were observed and compared before and after treatment.</p><p><b>RESULTS</b>The efficacy on TCM symptoms was obviously better in the treated group than that in the control group (P<0.01), and there were statistically remarkable decreases in aspect of FBG, 2h PBG, HbA1c and CSQS in the treated group as compared to those in the control group (P<0.05 or P<0.01), while no significant difference was found in BMI, hepatic and renal functions between the two groups (P>0.05).</p><p><b>CONCLUSION</b>The combined therapy of TFGs with sulfonylureas hypoglycemic drug could lower the blood glucose level and ameliorate clinical symptoms in the treatment of T2DM, and the therapy was relatively safe.</p>

Diagnosis, treatment and gene mutation analysis of the first case with dihydropteridine reductase deficiency in the mainland of China / 中华儿科杂志

<p><b>OBJECTIVE</b>The 6-pyruvoyl-tetrahydropterin synthase (PTPS) deficiency is the most common type of tetrahydrobiopterin (BH4) deficiency. The reported patients with BH4 deficiency are all PTPS deficient found in the mainland of China previously. The activity of dihydropteridine reductase in BH4 metabolism has been determined for 902 patients with hyperphenylalaninemia in the authors' laboratory since 2003. The purposes of this study were to characterize the first case with DHPR deficiency who was diagnosed in June, 2007, to investigate the clinical manifestation, the differential diagnostic criteria, the effect of treatment as well as gene mutation of DHPR deficiency.</p><p><b>METHODS</b>(1) A male patient presented with poor hand control, seizure, hypotonia and mental retardation since five-month after birth. His phenylalanine (Phe) level was 600 micromol/L and he was diagnosed as hyperphenylalaninemia at the age of one year and six-month. (2) This patient was subjected to combined Phe (100 mg/kg) and BH4 (20 mg/kg) loading test, to evaluate the degree of Phe level response to BH4. Urinary neopterin and biopterin analysis as well as the determination of DHPR activity in dried blood spot were also performed. (3) The blood DNA samples of the patient and his parents were collected to amplify the seven exons of QDPR gene using related primers, and the amplified products were directly sequenced for mutation analysis. (4) The patient was treated with BH4 or with a combined small amount of Phe-free special milk, neurotransmitter precursors and folic acid after the diagnosis and was followed up for clinical effects of treatment.</p><p><b>RESULTS</b>(1) The basic Phe level was 476 micromol/L, then it increased to 1355 micromol/L at 3 h after taking Phe and slowly decreased to 610 micromol/L at 24h after taking BH4. (2) The basic urinary neopterin and biopterin were 2.92 mmol/mol Cr (normally < 2.61 mmol/mol Cr) and 7.44 mmol/molCr (normally < 2.67 mmol/mol Cr) respectively, and biopterin percentage was 71.79% (normally 42.7% - 75.9%). The patient had higher biopterin level. (3) The DHPR activity of this patient was (0.27 - 0.51) nmol/(min.5 mm disc) which were 6.11% - 10.6% of normal control, so he was diagnosed as DHPR deficiency. (4) The analysis of QDPR gene mutation showed that the patient carries missense mutation c.515C > T (P172L) from his father and nonsense mutation c.661C > T (R221X) from his mother. The c.515C > T is not reported before, we also did not find this mutation in 50 normal children. (5) The patient started to be treated with large dosage of BH4 (10 - 20) mg/(kg.d) or BH4 combined with small amounts of Phe-free milk, neurotransmitter precursors L-dopa (3 - 5) mg/(kg.d) plus carbidopa, 5-hydroxytryptophan (3 - 5) mg/(kg.d), and folic acid 15 mg/d as well at the age of one year and six-month after the diagnosis. The seizure has disappeared, the symptoms such as hypotonia have been obviously improved and the Phe level was 60 micromol/L at the six months after the treatment in this patient.</p><p><b>CONCLUSION</b>(1) The patient with DHPR deficiency has common symptoms of BH4 deficiency (such as fair hair, hypotonia, mental retardation), and there is metabolic disturbance of folic acid in DHPR deficiency. (2) The higher Phe levels slowly decreased after BH4 loading test, the urinary biopterin level was very high and the DHPR activity was very low in the patient with DHPR deficiency. (3) The c.515C > T may be a new mutation of QDPR gene. (4) The DHPR deficient patient must be treated with higher dose of BH4 (8 - 20) mg/(kg.d), neurotransmitter precursors and folic acid as well.</p>