Bone Neoplasms ; Female ; Humans ; Middle Aged ; Waldenstrom Macroglobulinemia

The culture conditions of Saccharomyces cerevisiae sp.strain by 1.1 b were optimized for the production of D-(-)-mandelate dehydrogenase which is useful for the asymmetric bioreduction of benzoylformate to form D-(-)-mandelate.The optimum medium(per liter)consistes of 60 g peptone,30 g maltose, 0.5 g MgSO_4,0.01 g ZnSO_4,1.0 g KCl.After optimization of the culture medium,the enzyme production in shake flasks is enhanced from 2.56 to 20.21 U/L.The optimum fermentation conditions were determined as follows:medium volume 100 mL(i.e.,40%for a 250-mL shake flask),pH 6.5,inoculum size 10%,temperature 30℃,and cultivation time 25 h.

The development status of standardization of Chinese medicine and acupuncture in Australia and New Zealand is respectively introduced from 3 levels-national standard, regional standard and association standard. A national registration standard for Chinese medicine has been implemented since July 1, 2012 in Australia. The Oceania Federation of Chinese Medicine and Acupuncture Societies was also founded in capital of New Zealand. Four characteristics are revealed from researches and analyses: people's needs and the relevant system are the foundations of national standards of Chinese medicine; legislation on Chinese medicine is the guarantee for setting and implementing national standards, where necessity, scientificity, vitality, diversity and breakthroughs are embodied; registration standards are the key in international standardization of Chinese medicine; and international organizations are major force in promoting standardization of Chinese medicine and acupunc ture.
Acupuncture Therapy ; standards ; Australasia ; Drugs, Chinese Herbal ; standards ; Humans ; Medicine, Chinese Traditional ; standards ; Oceania ; Reference Standards

Objective To investigate the incidence and relevant information of preterm birth and the outcomes of preterm infants delivered at various gestational weeks and for different causes. Methods Totally 955 women, who ended their pregnancies before term, and 1066 neonates of the previous mothers were enrolled in this survey, among 15 197 deliveries at Peking University First Hospital, Beijing Gynecological and Obstetric Hospital, Women's and Children's Hospital of Haidian District and Peking University Third Hospital, respectively, from December 1~(st), 2006 to May 31~(st), 2007. Results (1)Incidence of preterm birth: The overall incidence of preterm birth of the 4 hospitals was 6. 3% (955/15 197), and it was 8.1% (125/1549) in Peking University First Hospital, 13.1% (150/1142), which was the highest (P<0.01), in Peking University Third Hospital, 5.5% (369/6656) in Beijing Gynecological and Obstetric Hospital and 34.0% (311/5850) in Women's and Children's Hospital of Haidian District.The preterm birth rate at the two comprehensive hospitals was significantly higher than that of the two specialized hospitals [10.2% (275/2691) vs 5.4% (680/12 506), P <0.01]. (2) Gestational weeks at delivery: The incidence of preterm birth before 34 weeks was 28.5% (272/954) and the number changed to 71.5% (682/954)for those preterm deliveries after 34 weeks. However, this number varied among the 4 hospitals. Peking University First Hospital had the highest incidence of preterm birth before 34 weeks(P< 0.05), and the lowest was found in Women's and Children's Hospital of Haidian District(P<0.01), but no difference was found between Peking University Third Hospital and Beijing Gynecological and Obstetric Hospital. (3) Etiology of preterm birth: Preterm premature rupture of membranes (PPROM) accounted for the most proportion of all preterm birth cases, followed by iatrogenic preterm birth and spontaneous preterm birth. But the causes of preterm birth in the 4 hospitals were different. Peking University Third Hospital had a higher incidence of iatrogenic preterm birth than the others (P<0.01), and Peking University First Hospital had a higher incidence of preterm birth caused by PPROM and lower incidence of spontaneous preterm birth. The first four reasons of iatrogenic preterm birth were preeclampsia (143, 42.0%), fetal distress (58, 17.1%), placenta previa (43, 12.6%) and placenta abruption (33,9.7%). (4) Neonatal outcomes in different hospitals: The neonatal outcomes were quite different among the 4 hospitals due to different causes and different delivery weeks. The highest neonatal mortality rate was found in Beijing Gynecological and Obstetric Hospital (5.4%, 22/408) compared to that in Women's and Children's Hospital of Haidian District (1.3%,4/320) and Peking University Third Hospital (0. 6%, 1/170) (P< 0.01), but without any difference when compared to that in Peking University First Hospital (2.4%, 3/ 124) (P>0.05). (5) Neonatal outcomes at different gostational age: The recovery rate of preterm infants delivered at <32 weeks was lower than those delivered ≥32 weeks (P<0.01), and this number rose to 99. 6% in those delivered ≥34 weeks. More infants delivered <32 weeks were given up for treatment or died during the perinatal period than those delivered ≥32 weeks, with the neonatal mortality rate of 22.1% for those delivered at <32 weeks and only 0.3% for those delivered at ≥ 34 weeks (P<0.01). (6) Neonatal outcomes for various causes: The premature neonatal mortality rate for iatrogenic preterm births was higher than that of PPROM (4.9% vs 1.6%, P<0.05). But the neonatal recovery rates were similar among the PPROM, spontaneous and iatrogenic preterm birth group (P>0.05). Conclusions Preterm birth is associated with high perinatal mortality rate, especially for those delivered before 32 weeks which would be highlighted in prevention. Reduction of the iatrogenic preterm birth, combined with proper prevention of PPROM, is an important issue in decreasing the prevalence of preterm birth.

OBJECTIVETo construct an expression vector of siRNA targeting human MTA1 gene and observe its gene-silencing effect in esophageal carcinoma cells.

METHODSThe siRNA sequences targeting MTA1 gene were designed and synthesized with two complementary oligonucleotide strands. The oligonucleotide strands were annealed and recombined into pRNAT-U6.2 vector, which was identified by sequencing following transformation and amplification. The siRNA expression vector pRNAT-U6.2-MTA1 was transfected into human esophageal carcinoma EC9706 cells via liposome. RT-PCR and Western blotting were used to detect expression levels of MTA1 mRNA and protein in the transfected EC9706 cells, respectively.

RESULTSThe double-stranded oligonucleotide fragments of the siRNA targeting MTA1 gene were cloned into pRNAT-U6.2 vector, which was validated by sequence analysis. RT-PCR and Western blotting indicated that MTA1 mRNA and protein expressions were significantly decreased in the transfected cells, especially in those transfected with the siRNA targeting the sequence of GACCCTGCTGGCAGATAAA (481-499), which induced almost complete silencing of MTA1 protein expression.

CONCLUSIONThe siRNA expression vector pRNAT-U6.2-MTA1 for silencing MTA1 gene expression in the esophageal carcinoma cells has been successfully constructed, which may facilitate further study for decreasing the invasive and metastatic potentials of malignant tumors by MTA1 gene silencing.

Base Sequence ; Blotting, Western ; Cell Line, Tumor ; Cloning, Molecular ; Esophageal Neoplasms ; genetics ; metabolism ; pathology ; Genetic Vectors ; genetics ; Histone Deacetylases ; biosynthesis ; genetics ; Humans ; Molecular Sequence Data ; RNA Interference ; RNA, Messenger ; biosynthesis ; genetics ; RNA, Small Interfering ; genetics ; Repressor Proteins ; biosynthesis ; genetics ; Reverse Transcriptase Polymerase Chain Reaction ; Transfection

Objective The factors influencing the dose distribution of intracavitary brachytherapy for moderately advanced and advanced uterine cervical cancer was studied.Methods Ninty-five patients with cervical cancerⅡ～Ⅲb who received radical radiation therapy in our department from Aug,2004 to Nov,2005,were treated with after-loading brachytherapy using,first,the self-designed“Mutipurpose Hori- zontal Transit Table”(MPHTT) for locating and treatment before the intracavitaray brachytherapy proper. The deviation of isodose curve based on A-B reference system,and the dose of deviation was defined by measuring in a practical standard phantom.Results There were significant influence on the deviation of i- sodose curve in pathology and para-metrial infiltration of cervical cancer and operating skill,but negative to clinical stage.The degree of deviation of isodose curve could not be lowered with the increase in sessions of intracavitary brachytherapy.Conclusions It is necessary to perform the locating,by use of mphtt,before the proper brachytherapy for patients with cervical cancer,not only for the identification of the deviation of i- sodose curve,but also to provide the evidence for revising the plan for dose adjustment of conformal radiation therapy in the pelvic cavity.

Animals ; Female ; Guinea Pigs ; Kidney ; microbiology ; pathology ; Leptospira interrogans ; isolation & purification ; pathogenicity ; Leptospirosis ; microbiology ; pathology ; Liver ; microbiology ; pathology ; Lung ; microbiology ; pathology ; Male ; Time Factors

OBJECTIVETo explore the molecular mechanisms of arsenic trioxide (As2O3) inhibiting NB4 cells proliferation.

METHODSThe Janus kinase 1 (JAK1) protein level and its phosphorylation level in NB4 cells was detected by Western blots. NB4 cells were transfected with JAK1 siRNA or JAK1 plasmid to make JAK1 gene silenced or overexpressed. The inhibition of NB4 cells proliferation was measured by MTT assay and Trypan blue exclusion respectively. The variation of phosphorylation level of JAK1 and the cell cycle inhibitor P21 were determined by Western blots.

RESULTSJAK1 protein was expressed stably in NB4 cells, with no phosphorylation. The phosphorylation of JAK1 was enhanced after the NB4 cells treated with As2O3. After NB4 cells transfected with JAK1 siRNA, the expression level of JAK1 was obviously lower than that of in the non-specific siRNA group and blank control group. The effect of As2O3 inhibiting NB4 cells proliferation was weaker in the JAK1 siRNA transfected group. The inhibiting rate of 4 micromol/L As2O3 on NB4 cells proliferation of JAK1 siRNA group was 49.12% being lower than that of the non-specific siRNA group (74.58%) and control group (72.33%). After NB4 cells transfected with JAK1 plasmid, the JAK1 expression level in wild-type and mutant type plasmid groups were significantly higher than those in the empty plasmid group, moreover the effect of As2O3 inhibiting proliferation was stronger in wild-type plasmid group. The inhibiting rate of 4 micromol/L As2O3 on NB4 cells proliferation of wild-type plasmid group was 69.53% being higher than that of the mutant type JAK1 plasmid group (37.26%) and the empty plasmid group (39.61%). The expression level of P21 was up-regulated after the NB4 cells treated with As2O3.

CONCLUSIONJAK1 is expressed stably in NB4 cells, but has no activity. Arsenic trioxide inhibits the proliferation of NB4 cells through activating the JAK1. P21 is up-regulated after arsenic trioxide activated the JAK1 to inhibit the proliferation of NB4 cells.

Apoptosis ; drug effects ; Arsenicals ; pharmacology ; Cell Proliferation ; drug effects ; Humans ; Janus Kinase 1 ; genetics ; metabolism ; Oxides ; pharmacology ; Signal Transduction ; drug effects ; Tumor Cells, Cultured

OBJECTIVETo isolate, identify and culture human spermatogonial stem cells (SSC) and then obtain purified and enriched human SSCs for research and application.

METHODSWe detected the expression of CD90 in the human testis using the immunofluorescence technique and isolated human testicular spermatogenic cells by two-step enzymatic digestion, followed by differential plating and magnetic-activated cell sorting (MACS) with CD90 as an SSC marker. Then we identified the isolated CD90-positive spermatogenic cells by RT-PCR and immunocytochemistry, and meanwhile cocultured them with Sertoli cells in SG medium in vitro.

RESULTSThe isolated CD90-positive cells showed a relatively homogeneous characteristic in size and morphology and expressed the genes specific for human SSCs, with high expressions (90.5%) of GFRA1, GPR125, and UCHL1. After coculture with Sertoli cells in the SG medium for 2 weeks, the isolated CD90-positive cells maintained a good activity.

CONCLUSIONCD90 can be regarded as a speci- fic marker for human SSCs and used to obtain highly enriched human SSCs by differential plating and MACS. Furthermore, the isolated human SSCs can be cultured in SG medium in vitro.

Adult Stem Cells ; cytology ; Biomarkers ; metabolism ; Cell Separation ; methods ; Cell Shape ; Cell Size ; Coculture Techniques ; Glial Cell Line-Derived Neurotrophic Factor Receptors ; metabolism ; Humans ; Immunohistochemistry ; Male ; Receptors, G-Protein-Coupled ; metabolism ; Sertoli Cells ; Spermatogonia ; cytology ; Testis ; metabolism ; Thy-1 Antigens ; isolation & purification ; metabolism ; Ubiquitin Thiolesterase ; metabolism

Mesenchymal stem cells (MSC) are the ideal adult stem cells in cell/gene therapy and tissue engineering for their features of easily-handling, highly proliferative capacity in vitro, low immunogenicity and immunomodulatory ability. MSC, as a kind of cellular drug, have been utilized in a phase III clinical trial to treat refractory graft-versus-host disease (GVHD). However, the essences of MSC in culture remain elusive so far. Whether the cells expanded in vitro are stem cells per se, and if not, why expanded MSC maintain their multiple differentiation ability? And are the MSC cultivated in vitro homogeneous and if not, what the heterogeneity stands for? Focusing on the heterogeneity of MSC in culture in vitro, the above questions are briefly discussed in this review.
Cell Differentiation ; Cell Proliferation ; Cells, Cultured ; Humans ; Mesenchymal Stromal Cells ; cytology