Objective To investigate the clinical effectiveness of disease-syndrome combination therapy for anovulatory infertility through randomized controlled trial, thus to provide high-level evidence-based medical reference for the diagnosis and treatment of WHO typeⅡovulation disorder infertility. Methods Eighty-six patients with WHO typeⅡovulation disorder infertility were randomized into treatment group and control group , 43 cases in each group. Both groups were given the treatment for correcting endocrinic disorder if necessary. Additionally, by following the principle of combination of disease differentiation and syndrome differentiation , Zhuyun Recipe 1, Zhuyun Recipe 2, and Zhuyun Recipe 3 were separately given orally to the patients with kidney-yang deficiency (WHO type Ⅱ ovulation disorder without PCOS), patients with liver-kidney insufficiency(WHO typeⅡovulation disorder complicated with hyperandrogenitic lean-PCOS),and patients with kidney-deficiency complicated with phl egm-dampness (WHO type Ⅱ ovulation disorder complicated with insulin-resistant obesity-PCOS) in the treatment group respectively. The patients in the control group was given oral use of the first-rate ovulation induction drug Clomiphene. The treatment covered 3 menstruation cycles. Ovulation rate,pregnancy rate and alive fetus rate were used as the indicators for evaluating the effectiveness,and the safety was also monitored. Results(1)Of the 86 cases,5 cases dropped out(loss to follow-up or being excluded),with the drop-out rate being 6%. Of the 5 drop-out cases,3 were from the control group,and 2 were from the treatment group. (2)Ovulation rate of the treatment group was 70.7%,and that of the control group was 85.0%. The ovulation rate of the two groups was enhanced,but the difference between the two groups was insignificant(P > 0.05).(3)Pregnancy rate of the treatment group was 51.2%,and that of the control group was 30.0%, the differen ce being significant between the two groups(P < 0.05).(4)Alive fetus rate of the treatment group was 45.2%,and that of the control group was 22.5%,the difference being significant between the two groups(P < 0.05).(5)No adverse reaction was found in the two groups during the treatment. Conclusion The disease-syndrome combination therapy for WHO type Ⅱanovulatory infertility shows the advantages of higher pregnancy rate and alive fetus rate, which makes up for the disadvantages of low pregnancy rate and high abortion rate of Clomiphene.

OBJECTIVETo explore the effect of Salvia miltiorrhiza monomer IH764-3 on apoptosis in hydrogen peroxide (H2O2)-stimulated hepatic stellate cells (HSCs).

METHODSHSCs were cultured in medium with different IH764-3 doses (10 mg/L, 20 mg/L, 30 mg/L, 40 mg/L) and without IH764-3. Direct cell count, 3H-thymidine incorporation, Annexin-V/Propidium Iodide double-labeled flow cytometry, TUNEL and transmission electron microscopy were employed to estimate the influence of IH764-3 on proliferation and apoptosis of HSCs. The expression of extracellular signal-regulated kinase 1 (ERK1) mRNA and protein in HSCs were detected using RT-PCR and Western blot respectively.

RESULTSIt was showed that H2O2 could promote HSC proliferation. In contrast, IH764-3 at concentrations of 10 mg/L, 20 mg/L, 30 mg/L and 40 mg/L inhibited its proliferation. The inhibition rates were 7.13%, 28.36%, 53.80% and 73.10% (P < 0.01). And the inhibition rates of IH764-3 at concentrations of 30 mg/L at 12 h, 24 h and 48 h were 22.24%, 40.51% and 61.65%. Furthermore, IH764-3 could also induce the HSC apoptosis in dose-dependent an dtime-dependent manners (P < 0.01). In addition, after exposed of HSCs to IH764-3 for 24 h, ERK production decreased and ERK1 mRNA was down-regulated earlier about 2 h after exposure to IH764-3.

CONCLUSIONIH764-3 may inhibit the proliferation and induce apoptosis of HSCs in both dose-dependent and time-dependent manners, which may be related to down-regulation of ERK expression.

Apoptosis ; drug effects ; physiology ; Cell Line ; Down-Regulation ; drug effects ; Drugs, Chinese Herbal ; isolation & purification ; pharmacology ; Hepatic Stellate Cells ; cytology ; Humans ; Hydrogen Peroxide ; pharmacology ; Mitogen-Activated Protein Kinase 3 ; genetics ; metabolism ; RNA, Messenger ; genetics ; metabolism ; Salvia miltiorrhiza ; chemistry

BACKGROUNDPatients with myelodysplastic syndrome (MDS) display a very diverse pattern. In this study, we investigated prognostic factors and survival rate in adult patients with MDS refractory anaemia (MDS-RA) diagnosed according to French-American-British classification and evaluated the International Prognostic Scoring System (IPSS) for Chinese patients.

METHODSA multi-center study on diagnosis of MDS-RA was conducted to characterize the clinical features of Chinese MDS patients. The morphological criteria for the diagnosis of MDS-RA were first standardized. Clinical data of 307 MDS-RA patients collected from Shanghai, Suzhou and Beijing from 1995 to 2006 were analyzed using Kaplan-Meier curve, log rank and Cox regression model.

RESULTSThe median age of 307 MDS-RA cases was 52 years. The frequency of 2 or 3 lineage cytopenias was 85.6%. Abnormal karyotype occurred in 35.7% of 235 patients. There were 165 cases (70.2%) in the good IPSS cytogenetic subgroup, 44 cases (18.7%) intermediate and 26 cases (11.1%) poor. IPSS showed 20 (8.5%) categorized as low risk, 195 cases (83.0%) as intermediate-I risk and 20 cases (8.5%) as intermediate-II risk. The 1-, 2-, 3-, 4- and 5-year survival rates were 90.8%, 85.7%, 82.9%, 74.9% and 71.2% respectively. Fifteen cases (4.9%) transformed to acute myeloid leukaemia (median time 15.9 months, range 3 - 102 months). Lower white blood cell count (< 1.5 x 10(9)/L), platelet count (< 30 x 10(9)/L) and cytogenetic abnormalities were independent prognostic factors by multivariate analysis, but age (= 65 years), IPSS cytogenetic subgroup and IPSS risk subgroup were not independent prognostic factors associated with survival time.

CONCLUSIONSChinese patients were younger, and had lower incidence of cytogenetic abnormalities, more severe cytopenias but a more favourable prognosis than Western patients. The major prognostic factors were lower white blood cell count, lower platelet count and fewer abnormal karyotypes. The international prognostic scoring system risk group was not an independent prognostic factor for Chinese myelodysplastic syndrome patients with refractory anaemia patients.

Adolescent ; Adult ; Aged ; Aged, 80 and over ; Anemia, Refractory ; etiology ; mortality ; Asian Continental Ancestry Group ; Child ; China ; Female ; Humans ; Male ; Middle Aged ; Myelodysplastic Syndromes ; complications ; mortality ; Prognosis

Objective To investigate the prevalence and major risk factors of peripartum thromboembolic disease in different regions of Guangdong province.Methods Data from 169 218pregnant women in different regions of Guangdong province from January 2005 to June 2010 were analyzed retrospectively.The prevalence and epidemiological characteristics of thromboembolic disease during pregnancy or puerperium were investigated.Results Of the studied population,( 1 )20 l cases ( 1.3‰ ) suffered from thromboembolic disease during pregnancy or puerperium including 128 cases of deep vein thrombosis (DVT),68 cases of cerebral venous thrombosis (CVT) and 5pulmonary embolism,the prevalence rates were 0.8‰,0.4‰,and 0.02‰ respectively.(2) Risk factors in different regions showed that,in the Pearl River Delta area,the major risk factors for DVT would include previous or family history of thrombosis,pregnancy complications,with medically involved diseases,prolonged bed rest and pregnancy weight gain ＞ 15 kg etc.While in castern,western,northern parts of Guangdong,the major risk factors for DVT would include pregnancy weight gain ＞ 15 kg,prolonged bed rest,preeclampsia,cesarean section and complications during pregnancy.In Pearl River Delta region,the major risk factors for CVT would include eclampsia,preeclampsia,pregnancy complications,prolonged bed rest ＞3 days,past history or family history of thrombosis.While eclampsia,preeclampsia,advanced age or younger age,pregnancy weight gain ＞15 kg,complications during pregnancy were the major risk factors for CVT in the eastern,western or northem parts of Guangdong.Conclusion Prevalence and major risk factors of peripartum thromboembolic disease in different regions of Guangdong were different.It was crucial to take effective measures in pregnant women with different epidemiological characteristics and risk factors to prevent and reduce the incidence of peripartum thromboembolic disease.

OBJECTIVETo investigate hantanvirus infection of captured rodents in Haidian district and Changping district of Beijing and to type hantavirus using molecular technique.

METHODSThe captured mice were classified and the density of distribution was calculated. Reverse transcription-polymerase chain reaction (RT-PCR) technique was used to amplify the partial M fragnments of hantaviruse. Several representative positive samples were sequenced and analysed by ClustalX (5.0) and DNAClub software.

RESULTSA total of 414 animals were captured, among which Battus norvegicus was the dominant group. In Haidian district, the median infection rates with hantavirus were 13.14% in Battus norvegicus and 0 in Mus musculus Linnaeus. In Changping district, the average infection rates were 17.46% in Battus norvegicus and 3.57% in Mus musculus Linnaeus. Nucleotide sequences analysis showed that the virus detected all belonged to SEO-type. They clustered with Z37 virus and could be branched into 2 different subclades.

CONCLUSIONThe major hosts of hantavirus in Haidian and Changping district were Battus norvegicus and the epidemic strains in the two districts of Beijing were genotyped as SEO-type. Nucleotide sequence and deduced amino acid sequence from different rodents were highly homologous, while nucleotide mutation had also been observed. Further studies are required to explore the possible virus sequence mutation.

Animals ; China ; epidemiology ; DNA, Viral ; genetics ; Disease Reservoirs ; Fluorescent Antibody Technique ; Hantavirus ; classification ; genetics ; isolation & purification ; Hantavirus Infections ; epidemiology ; veterinary ; virology ; Hemorrhagic Fever with Renal Syndrome ; epidemiology ; veterinary ; virology ; Mice ; Molecular Epidemiology ; Phylogeny ; Rats ; Reverse Transcriptase Polymerase Chain Reaction ; Rodent Diseases ; epidemiology ; virology

OBJECTIVETo analyze a Duchenne muscular dystrophy(DMD) patient's muscular regeneration, dystrophin expression and locomotive variation before and after he underwent umbilical cord blood stem cell transplantation in order to assess the therapeutic effect.

METHODSA 12-year-old DMD boy who could not walk for 3 years was confirmed by gene analysis and dystrophin protein immune test on his muscle. He had no other chronic disease. By HLA matching, a piece of umbilical cord blood stem cell with 6 HLA sites matching to the boy was found in Guangdong Umbilical Cord Blood Bank. The number of the nucleated cells of the umbilical cord blood stem cell was 24.08x 10(8). After pretreatment for the DMD boy with busulfan, cyclophosphamide and rabbit anti-human thymocyte globulin, the allergenic cord blood stem cells were transplanted into him by intravenous injection. Cyclosporin A, methylprednisolone, MMF, prostaglandin E1 and ganciclovir were given after the transplantation. At the same time, Gran, the granulocytic cell stimulating factor, and gamma globulin were administered. The biochemistry profile including serum creatine kinase (CK), the reconstruction of blood making, the deletion exon of DMD gene, the regenerating muscles, the dystrophin protein expression, and the locomotive function of the DMD boy were tested regularly.

RESULTS(1) The white blood cells (WBC) of peripheral blood decreased gradually to zero after pretreatment. In a period of 15 days after transplantation, the neutrophil increased to 0.5x 10(9)/L; at 25 days, WBC increased to normal level. Blood platelet was more than 20x 10(9)/L at 22 days. The hemoglobin rose to 85-100 g/L. At 140 days, sternal puncture revealed the rapid growth of neutrophil, blood platelet and hemoglobin. (2)At 140 days, the blood type of the DMD boy transformed from type O to type AB (the donor's blood type being AB). There was no grafe versus host reaction. (3) At 18, 30, 43, 55, 74 and 233 days after transplantation, the PCR-short tandem repeat test of the boy's peripheral blood DNA showed that his genotype was completely the same as the donor's. The results of PCR-short tandem repeat tests of the bone marrow cells DNA by sternal puncture at 140, 183 and 235 days were the same as those of the blood DNA. (4) At 60 days, DMD gene analysis by PCR showed that the defected DMD gene (exon 19 deletion) had been corrected by the umbilical cord stem cells transplantation. (5) At 75 days, the biopsy of calf muscle showed there were myoblast cells and muscular tubes growing. The dystrophin expressions were weak, but a few of them were strong. DNA analysis showed that the donor's gene DNA accounted for 1%-13%. At 126 days, obviously increased dystrophin positive muscular fibers of the boy were found. The donor's fibers rose to 2.5%-25%. (6) The serum CK of the boy declined from 5735 U/L to 274 U/L. (7) At 100 days, physical examination revealed improvement in his arms and legs.

CONCLUSIONThe therapy of Duchenne muscular dystrophy with allogeneic umbilical cord blood hematopoietic stem cell transplantation may reset up the blood-making function, decrease the serum CK level, restore the dystrophin in muscles, and improve the locomotive function of the DMD boy. These data suggest that the allogeneic umbilical cord blood hematopoietic stem cell transplantation may benefit the DMD boys.

Alprostadil ; therapeutic use ; Busulfan ; therapeutic use ; Child ; Combined Modality Therapy ; Cord Blood Stem Cell Transplantation ; methods ; Cyclosporine ; therapeutic use ; Dystrophin ; genetics ; Ganciclovir ; therapeutic use ; Humans ; Male ; Methylprednisolone ; therapeutic use ; Muscular Dystrophy, Duchenne ; genetics ; therapy ; Polymerase Chain Reaction ; Treatment Outcome

OBJECTIVE: To establish a novel method to isolate endothelial progenitor cells（EPC） from cryopreserved umbilical cord blood (cryoUCB), to investigate the biological characteristics of EPC and to improve the rate of EPC obtained from cryoUCB. METHODS: Twelve cryoUCB samples during 2000 to 2001 years were collected from allogeneic cord blood bank, cryoUCB was thawed rapidly in a water bath at 37 ℃, total nucleated cells (TNCs) were washed by phosphate-buffered saline (PBS). TNCs were seeded onto fibronectin-coated dishes to isolate EPC. Flow cytometry and immunofluorescence were used to identify EPC. The function of EPC was identified in vitro, such as the incorporation of Dil-Ac-LDL and FITC-UEA-I, the formation of capillary-like structure in matrigel, and the release of VEGF by ELISA. RESULTS: One to five cluster of cobble stone-like cells appeared at 2-3 weeks after seeding. Flow cytometric analysis showed that positive rates of CD31, CD34, CD144, and VEGFR (CD309) were（92.91±5.20）%, （30.0±23.27）%, （88.55±3.83）% and （67.21±12.12）% in passage 1 to passage 3 of EPC. EPC could uptake Dil-Ac-LDL and FITC-UEA-I, form capillary-like network on Matriget and release VEGF. CONCLUSION EPC had been successfully isolated from cryopreserved umbilical cord blood by this method with high stability and reproducibility. EPC can be obtained in 85% frozen umbilical cord blood. This method may lay a foundation to supply abundant EPC for clinical application.
Cell Differentiation ; Cells, Cultured ; Endothelial Progenitor Cells ; Fetal Blood ; Reproducibility of Results ; Stem Cells

BACKGROUNDThere has been no a specific scale to measure quality of life (QOL) for prostate cancer patients receiving androgen deprivation therapy (ADT) to date. This study aimed to develop and initially validate the scale to evaluate QOL for prostate cancer patients receiving ADT.

METHODSThe scale was developed following international recommendations. Moreover, the items were all generated through literature review and referenced questionnaires. After being reviewed by expert panelists, the revised scale was formed and then completed by a convenience sample of 200 prostate cancer patients from our hospital. Explore factor analysis (EFA) was applied to test the construct validity, then split-half reliability, Cronbach's alpha, and test-retest reliability were applied to assess the reliability and stability of the scale.

RESULTSThe revised scale contained 22 items and a total of 200 participants had completed the scale. One hundred participants were randomly selected from the total 200 participants to perform EFA with varimax rotation on the revised scale, and "hot flashes" item was deleted for low factor loading. We selected only 3 items from each factor, then, the final scale was formed with 18-items. We selected another 100 participants to perform the EFA again on the final scale. It was demonstrated that the structure with 6 factors explained 72.5% of total variance and factor loading value was above 0.40 in all items of the factors. Moreover, the split-half reliability coefficient, Cronbach's alpha, and test-retest reliability coefficient were calculated to be 0.74, 0.63, and 0.89, respectively, exhibiting good reliability on the whole.

CONCLUSIONSThe scale was identified to be a valid and reliable instrument to measure QOL for prostate cancer patients receiving ADT. Moreover, further research is needed to overcome the potential drawbacks.

OBJECTIVEThis study was to expand the cytotoxic T lymphocytes (CTL) through inducing the differentiation of umbilical blood monomuclear cells (UBMNC) by using various combination of cytokines, and to investigate the functions of expanded CTL.

METHODSThe MNC were isolated by ficoll density gradient centrifugation. Then, the PHA-P, IFN-γ combined with IL-2, IL-15 and other cytokines were used for induction and expansion of the cord blood-derived CTL. The biological function of CTL was examined by phenotype analysis, cytotoxic tests and real-time fluorescence quantitative PCR.

RESULTSAfter expansion for 15 days, the cell number increased by 1522% ± 137%. The content of CD3(-)CD8(-) cells in uncultured cord blood MNC was 95%, and the CD3(+)CD8(+) CTL cells reached 82.77% in cultured cord blood MNC after expansion for 15 days. The expanded CTL cell showed the cytotoxic activity against K562 and HeLa cell line. The killing rate of MNC was 61.88 ± 1.08%. After expansion, the killing rate could reach to 90% with the average value of 90.33 ± 2.02%. The expanded CTL cells highly expressed some key cytokines, such as granzyme A, granzyme B, GM-CSF, granulysin, IFN-γ, TGF-β, TNF-α and perforin. Compared with the control group, the expression of IFN-γ and TGF-β significantly increased (P < 0.05), and the other factors dramatically increased (P < 0.01).

CONCLUSIONThe cord blood-derived CTL can be expanded by different combinations of cytokines. These protocols may provide alternative choices for CTL cell expansion in tumor adoptive immunotherapy.

Cytokines ; Fetal Blood ; Granulocyte-Macrophage Colony-Stimulating Factor ; Granzymes ; Histocompatibility Antigens Class I ; Histocompatibility Antigens Class II ; Humans ; Immunotherapy, Adoptive ; Perforin ; Phytohemagglutinins ; T-Lymphocytes, Cytotoxic

OBJECTIVE: To explore the clinical effect of warming-needle moxibustion with different lengths of moxa stick for asthenospermia with kidney deficiency and liver depression. METHODS: A total of 240 patients with asthenospermia of kidney deficiency and liver depression were randomly divided into a 4-cm group (moxibustion with 4-cm moxa stick, 60 cases, 3 cases dropped off), a 3-cm group (moxibustion with 3-cm moxa stick, 60 cases, 4 cases dropped off), a 2-cm group (moxibustion with 2-cm moxa stick, 60 cases, 2 cases dropped off) and an acupuncture group (60 cases, 3 cases dropped off). All patients were treated with warming-needle moxibustion with different lengths of moxa stick or conventicnal acupuncture at Zhongwan (CV 12), Guanyuan (CV 4), Zhongji (CV 3), Guilai (ST 29), Yaoyangguan (GV 3), Guanyuanshu (BL 26), etc., once a day, five times a week; 4-week treatment was taken as one course, a total of two courses of treatment were given. The semen routine indexes, seminal plasma biochemical indexes, sex hormone levels and TCM syndrome score were compared before and after treatment among the 4 groups. RESULTS: After treatment, the sperm density, sperm viability, ratio of grade A sperm, ratio of grade A and B sperm, seminal plasma fructose and neutral α-glucosidase were higher than those before treatment (P<0.05, P<0.01), and the sperm deformity rates were lower than those before treatment in the 4-cm group and the 3-cm group (P<0.01, P<0.05). The ratio of grade A sperm, ratio of grade A and B sperm, seminal plasma fructose and neutral α-glucosidase in the 4-cm group were higher than the other three groups (P<0.05, P<0.01), and the sperm deformity rate was lower than the other three groups (P<0.05). After treatment, except for dizziness and tinnitus score, each-domain score and total scores of TCM syndrome scale in the 4-cm group and the 3-cm group were lower than those before treatment (P<0.01, P<0.05). The each-domain score of depression, weak waist and knees, low sexual function and total score in the 4-cm group were lower than those in the other three groups (P<0.05, P<0.01). The total effective rate was 87.7% (50/57) in the 4-cm group, which was higher than 78.6% (44/56) in the 3-cm group, 77.6% (45/58) in the 2-cm group and 70.2% (40/57) in the acupuncture group (P<0.05, P<0.01). CONCLUSION The warming-needle moxibustion with 4-cm moxa stick could effectively improve quality and motility of sperm and clinical symptoms in patients with asthenospermia of kidney deficiency and liver depression, which is superior to moxibustion with 3-cm, 2-cm moxa sticks and conventional acupuncture.
Acupuncture Points ; Acupuncture Therapy ; Depression ; Fructose ; Humans ; Kidney ; Liver ; Male ; Moxibustion ; Semen ; Treatment Outcome ; alpha-Glucosidases