In the present study, the specifically knockdown models of P-gp or MRP2 were constructed by using a series of chemically synthesized small interfering RNA (siRNA) in vitro. The expression of P-gp and MRP2 was measured by real-time PCR and Western blot, and the function was evaluated by applying P-gp and MRP2 substrate, rhodamine and methotrexate. The results showed that MRP2 siRNA-3 or P-gp siRNA-2 significantly decreased the mRNA expression of MRP2 or P-gp, the inhibition ratio was 68% or 84%; MRP2 siRNA-3 or P-gp siRNA-2 at a dose of 80 nmol x L(-1) significantly reduced the protein expression of MRP2 or P-gp at 48 h after treatment, the inhibition ratio was 62% or 70%. Meanwhile, other transporters were not influenced by siRNA. When pretreatment with MRP2 siRNA-3 or P-gp siRNA-2, the efflux of methotrexate or rhodamine decreased significantly and the intra-cellular concentration increased. The results suggested that chemically synthesized siRNA could significantly inhibit the expression and function of MRP2 and P-gp, and the model of RNAi in vitro could be used to evaluate the role of efflux transporters in transportation of drugs.
ATP-Binding Cassette, Sub-Family B, Member 1 ; genetics ; Gene Knockdown Techniques ; Multidrug Resistance-Associated Proteins ; genetics ; RNA Interference ; RNA, Small Interfering ; Real-Time Polymerase Chain Reaction

Adult ; Carcinoma, Small Cell ; metabolism ; pathology ; Diagnosis, Differential ; Female ; Follow-Up Studies ; Humans ; Hysterectomy ; Leukosialin ; metabolism ; Lymphoma ; metabolism ; pathology ; Peroxidase ; metabolism ; Sarcoma, Myeloid ; metabolism ; pathology ; surgery ; Uterine Cervical Neoplasms ; metabolism ; pathology ; surgery ; Uterine Cervicitis ; metabolism ; pathology

Objective To explore the expression of HCK gene during the cardiomyocyte differentiation of mouse embryonic stem cells and analyze the role of HCK gene in maintenance of pluripotency of embryonic stem cells.Methods Mouse embryonic stem cells were cultured,then induced to differentiate into cardiomyocytes.Total RNAs were isolated from mouse embryonic stem cells in the differentiation days:0 day(D0),the second day(D2),the fourth day(D4),the sixth day(D6),the eighth day(D8),respectively.The levels of HCK mRNAs were assessed by the method of semi-quantitive reverse transcriptase-polymerase chain reaction(RT-PCR).In the meanwhile,Total proteins were also isolated from mouse embryonic stem cells in the differentiation D0,D2,D4,D6,D8,and the levels of HCK proteins were evaluated by Western-blot.Results HCK mRNAs could be detected in the mouse embryonic stem cells in D0 and D2,however,they were undetectable from D4 to D8.The expression of HCK mRNAs was rapidly down-regulated during cardiomyocyte differentiation of mouse embryonic stem cells.Expression of HCK proteins,which coincided with HCK mRNAs,down-regulated during differentiation and couldn't be detected in D4.Conclusions With the cardiomyocyte differentiation of mouse embryonic stem cells,the expression of HCK in the levels of mRNA and proteins are sharply down-regulated;HCK may play an important role in maintaining the pluripotency of embryonic stem cell.

Purpose To summarize the clinicopathological features, immunophenotype, diagnosis and differential diagnosis of mammary analogue secretory carcinoma of salivary glands(MASC). Methods Four cases of MASC were included. The clinical, histological, and immunohistochemical features and follow-up data were evaluated. Results The patients aged between 20 to 72 years. Two cases were male and two were female. The average age was 52.2 years. All cases showed infiltrative growth and diverse cytology and histology, including lobular, cystic papillary, solid, cribriform or glandular structures, and microcystic honeycombed pattern composed of small cysts merged into larger cysts resembling thyroid follicles. There were no necrosis, nerve or vascular invasion in 4 cases. S-100 protein, Mammaglobin or SOX10 were strongly positive in all MASC cases.3 cases were insignificant positivity for GATA-3. Both p63 and DOG-1 were negative. Conclusion MASC is a lowgrade malignant epithelial tumor of salivary glands. Combined immunohistochemical positivity of S-100 protein, Mammaglobin and SOXIO, or negative for DOG-1, are useful in the diagnosis and differential diagnosis of MASC.

· AIM:To investigate the prevalence of juvenile myopia and factors affecting its occurrence in Linxia Prefecture.· METHODS:Totally 8 683 juvenile students who were 6-18 years old were extracted as respondent with stratified cluster sampling method.The eyesight,diopter and axial length were detected,and the gender,age,ethnicity,eye behavior were collected by self-made questionnaire.· RESULTS:The prevalence of myopia was 42.80％.The myopic rates of all ethnic groups were as follows:Tibetan Nationality:59.05％,Han Nationality:46.71％,Dongxiang Nationality:46.36％,Tu Nationality:45.24％,Sala Nationality:41.30％,Baoan nationality:40.61％,Hui Nationality:31.97％.Myopia rate between each ethnic groups had statistical difference (x2 =44.08,P=0.007).Multivariate analyses revealed that outdoor activities during the break was the protect factor for myopia;age,using electronic products on weekends and holidays,using the same lighting in room while studying were risk factors for myopia.· CONCLUSION:The prevalence of myopia increase with age in Linxia.The prevalence in different ethnic groups is different.Education department and family should make joint efforts to increase the outdoor time,improve the environment for teenagers to reduce the occurrence and development of myopia.

The megakaryocyte and platelet lineage may be one of the major sites of human cytomegalovirus (HCMV) infection. However, whether HCMV aggravates apoptosis in normal megakaryocytes was not well investigated. Megakaryocytic cell line CHRF-288-11 and HCMV AD 169 strain were co-cultured in this study. PCR was used to detect the direct infection of the cells by HCMV IEA expression. The apoptotic cells were analyzed by morphologic observation, DNA ladder formation, annexin V/PI and PI assay with flow cytometry. The results showed that HCMV significantly inhibited the growth of CHRF cells in three different concentrations of viral infection groups (10(-3), 10(-2), 10(-1)). The viability levels in each infection groups were 77%, 73% and 68% respectively after incubation for 7 days, compared with 98% in the control group. Using annexin V/PI with flow cytometry, it was shown that the percentages of apoptotic cells viral infection in groups (10(-3), 10(-2), 10(-1)) were (21.3 +/- 2.49)%, (25.8 +/- 3.65)% and (31.4 +/- 3.91)% at 7 days after infection, while the control was (3.68 +/- 1.47)%. The apoptotic cells were further confirmed by morphologic observation and DNA ladder formation. Furthermore, PCR detection also showed the direct infection by identification of HCMV IEA expression in CHRF cells. This study suggested that HCMV could directly infect megakaryocytes and aggravated apoptosis in HCMV-infected megakaryocytes.
Apoptosis ; Cell Survival ; Cells, Cultured ; Cytomegalovirus ; pathogenicity ; DNA, Viral ; analysis ; Humans ; Megakaryocytes ; cytology ; virology ; Polymerase Chain Reaction

OBJECTIVETo compare therapeutic effects of abdominal electroacupuncture (EA) and western medicine on poststroke constipation.

METHODSEighty cases were randomly divided into an EA group and a medication group, 40 cases in each group. The EA group were treated with EA at Daheng (SP 15), Fujie (SP 14), Tianshu (ST 25), Shuidao (ST 28), etc., once a day, 30 min each session, and the medication group with oral administration of 10 mg Cisapride, thrice each day. Seven days constituted one course. After 2 courses, clinical therapeutic effects were evaluated by cumulative scores of symptoms.

RESULTSThe total effective rate of 92.5% in the EA group was significantly better than 72.5% in the medication group (P < 0.05). After treatment, the cumulative scores of clinical symptoms significantly decreased in the two groups (P < 0.05) and the improving degrees of symptoms in the EA group was significantly better than that in the medication group (P < 0.05).

CONCLUSIONAbdominal electroacupuncture has a definite therapeutic effect on poststroke constipation, accelerating gastrointestinal movement.

Abdomen ; Aged ; Constipation ; etiology ; therapy ; Electroacupuncture ; Female ; Humans ; Male ; Medicine, Chinese Traditional ; Middle Aged ; Stroke ; complications

OBJECTIVETo investigate the effect of allogeneic leukocyte immunization combined with in vitro fertilization-embryo transfer (IVF-ET) for treatment of infertility induced by habitual abortion.

METHODSAllogeneic leukocyte immunization was performed in 9 patients with infertility induced by habitual abortion, with another 9 patients undergoing IVF-ET without habitual abortion as the control group. All the patients were treated with long GnRH-a protocols. The infertility patients with recurrent spontaneous abortion history were immunized with lymphocytes from the husband for before IVF-ET and after clinical pregnancy.

RESULTSThe fertilization rates of the immunotherapy group and control group were 81.3% and 82.2%, respectively, showing no significant difference (P>0.05). Five patients in each group had clinical pregnancy, and a twin pregnancy occurred in the control group. The embryo implantation rates were also comparable between the two groups (22.7% vs 28.6%, P>0.05). All the fetuses resulted from IVF-ET developed normally and were healthily delivered.

CONCLUSIONAllogeneic leukocyte immunotherapy along with IVF-ET is effective for treatment of infertility resulting from recurrent spontaneous abortion.

Abortion, Habitual ; physiopathology ; Adoptive Transfer ; methods ; Adult ; Embryo Transfer ; Female ; Fertilization in Vitro ; methods ; Humans ; Infertility, Female ; physiopathology ; therapy ; Pregnancy ; Pregnancy Outcome ; Treatment Outcome

OBJECTIVETo investigate the effect of moxibustion at Sanyinjiao (SP 6) for uterine contraction pain in labor, and evaluate the safety of the parturient and newborn.

METHODSOne hundred and seventy-four cases of singleton pregnancy and cephalic presentation primipara were single blinded and randomly divided into three groups: observation group (59 cases), placebo treated group (57 cases) and blank group (58 cases). The observation group was treated with moxibustion at Sanyinjiao (SP 6) for 30 min when the uterus cervix openning at 3 cm, the placebo treated group was treated with moxibustion at no acupoint for 30 min and the blank group was treated with routine labor nursing, the uterine contraction pain and the safety of the mother and infant were compared among three groups.

RESULTS1) The uterine contraction pain was tested by Visual Analogue Scale (VAS): the scores of VAS in the observation group were obviously decreased after 15 min and 30 min of moxibustion (both P<0.05), there were no obvious changes of the VAS scores in placebo treated group and the blank group, the scores of VAS in observation group decreased much more obviously than those in the other two groups (all P<0.05); 2) Midwife rating of the uterine contraction pain: after 30 min of moxibustion, the effective rate of labor analgesia was 69.5% (41/59) in observation group, which was higher than that of 45.6% (26/57) in placebo treated group and 43.1% (25/58) in blank group, with significant differences between them (both P<0.05); 3) The postpartum hemorrhage amount of the observation group was obviously lower than those of placebo treated group and blank group (both P<0.05); 4) The Apgar score of newborn was higher in observation group and placebo treated group than that of blank group (both P<0.05).

CONCLUSIONMoxibustion at Sanyinjiao (SP 6) can relieve the uterine contraction pain, and has no side effect to mother and infant, it is one of the safe, effective and simple non-drug analgesia methods.

Acupuncture Points ; Adult ; Female ; Humans ; Labor Pain ; physiopathology ; therapy ; Labor, Obstetric ; Moxibustion ; Pregnancy ; Treatment Outcome ; Uterine Contraction ; Young Adult

OBJECTIVETo study the effect of Banxia Houpu Decoction on a chronic mild stress model of depression and investigate the antidepressive mechanism.

METHODWith consumption of a 1% sucrose solution as an index, and by subjecting rats to a variety of mild stressors for a prolonged period of time, a chronic mild stress model was developed. The levels of the blood lipid were measured by blood lipid kits, the natural kill (NK) cell activity in the spleen was measured with the method of the enzyme lactate dehydrogenase (LDH) released, the superoxide dismutase (SOD) activity in red blood cell was assayed by the Autoxidation of Pyrogallol method, the NO synthase (NOS) activity in serum and tissue was measured by NOS kits, and the content of malondialdehyde(MDA) was measured by MDA kits.

RESULTBanxia Houpu Decoction significantly increased the consumption of sucrose solution, increased the level of the high density lipoprotein (HDL-C), decreased the level of the Triglyceride(TG) in serum, enhanced the activity of the NK in spleen, decreased the activity of the SOD in red blood and the activity of the NOS in serum and tissue, and reduced the content of MDA in tissue by effect on lipid Peroxidation in CNS model of depression.

CONCLUSIONBanxia Houpu Decoction has antidepressant effect in different ways.

Animals ; Antidepressive Agents ; pharmacology ; Depression ; immunology ; metabolism ; Drugs, Chinese Herbal ; pharmacology ; Killer Cells, Natural ; immunology ; Male ; Malondialdehyde ; blood ; metabolism ; Myocardium ; metabolism ; Rats ; Rats, Sprague-Dawley ; Stress, Psychological ; complications ; Sucrose ; metabolism ; Superoxide Dismutase ; metabolism ; Triglycerides ; blood