Pseudomonas sp. M18, one of plant-growth-promoting rhizobacteria, can produce secondary metabolites including phenazine-1-carboxylic acid (PCA) and pyoluteorin (Plt). PA2572 gene coding protein is a probable two-component response regulator in Pseudomonas according to homologous speculations. In order to investigate its genetic function, PA2572 homologous gene, ppbR, was amplified from M18 genome, inactivated by inserting a Gm cassette. The resulting reconstruct was introduced into the M18 genome using homologous recombination technique, so as to obtain the null mutant M18P. The results showed that the M18P has less flagellar swimming and swarming motility, and yielded fewer PCA. The production of PCA was only 50% of the wild type. However, there was no remarkable difference between mutant and wild type in producing pyoluteorin in KMB medium.

OBJECTIVETo evaluate the effects of levonorgestrel-releasing intrauterine system (LNG-IUS) combined with GnRH analogue (GnRH-a) in the treatment of adenomyosis with uterine body enlargement.

METHODSTwelve women (mane age 40.3 years) with adenomyosis and uterine cavity depth over 11 cm received injections of GnRH-a every 4 weeks, and after the uterine cavity depth was reduced to below 10 cm, LNG-IUS was deployed. VAS pain score, PBAC bleeding score, uterine volume, and hemoglobin levels of the women were measured before the treatment and at 6 and 12 months after LNG-IUS placement.

RESULTSThe VAS pain score was significantly lowered at 6 and 12 month after LNG-IUS placement (P<0.05), and the PBAC bleeding score also showed significant reductions (P<0.05). The uterine volume decreased significantly at 6 and 12 months after LNG-IUS placement as compared with that before the treatment, but was significantly greater at 6 month in comparison with that at the time of LNG-IUS placement (P<0.05). Serum hemoglobin levels underwent significant increments after LNG-IUS placement (P<0.05).

CONCLUSIONLNG-IUS combined with GnRH analogue injection can be effective in the treatment of adenomyosis with dysmenorrhea and hypermenorrhea.

Adult ; Delayed-Action Preparations ; administration & dosage ; Drug Therapy, Combination ; Endometriosis ; drug therapy ; Female ; Gonadotropin-Releasing Hormone ; analogs & derivatives ; therapeutic use ; Humans ; Levonorgestrel ; administration & dosage ; Uterine Diseases ; drug therapy

Adult ; Aged ; Female ; Humans ; Middle Aged ; Ovarian Neoplasms ; mortality ; pathology ; Prognosis ; Proportional Hazards Models ; Retrospective Studies ; Young Adult

OBJECTIVETo investigate the effect of rat mesenchymal stem cells (MSCs) transfected with different oncogenes differentiate into hepatocellular carcinoma (HCC) in vitro.

METHODSMSCs, transfected with different oncogenes c-myc, K-ras, c-myc and K-ras and amplified in vitro, were infused into rats via vena portae. The recipient rats were divided into the hepatic impairment group, which were fed with tetrachloromethane and the healthy control group. At day 7, 14, 21, 28 and 42 following grafting, the complete livers were obtained and examined using fluorescence detection, conventional pathology and immunohistochemistry detection of GFP, c-kit and AFP to study the colonization and distribution of stem cells in rat liver.

RESULTSNo immunological rejection occurred after grafting of allogenic MSCs. The infused MSCs colonized in the recipient rat liver. Liver tumors were present in 6 rats grafted with MSCs that were transfected with K-ras, K-ras and c-myc, and the protein expression of AFP was detected using immunocytochemistry at day 7. Rats grafted with MSCs that were transfected with c-myc gene had no obvious tuberosity or tumor. Small oval cells were found microscopically in the periphery of vena portae, and immunohistochemistry staining of AFP was negative. Immunohistochemical staining of c-kit was positive in all livers of rats that were transfected with MSCs.

CONCLUSIONSHepatocellular carcinoma may derive from genetically mutated MSCs.

Animals ; Bone Marrow Cells ; cytology ; metabolism ; Carcinoma, Hepatocellular ; metabolism ; pathology ; Cell Differentiation ; Cells, Cultured ; Female ; Genes, myc ; genetics ; Genes, ras ; genetics ; Genetic Vectors ; genetics ; Green Fluorescent Proteins ; genetics ; metabolism ; Liver ; metabolism ; pathology ; Liver Neoplasms, Experimental ; metabolism ; pathology ; Male ; Mesenchymal Stem Cell Transplantation ; Mesenchymal Stromal Cells ; cytology ; metabolism ; Microscopy, Fluorescence ; Rats ; Rats, Sprague-Dawley ; Transfection

OBJECTIVETo explore the deep pathogenesis of acquired cholesteatoma.

METHODSThe temporal bone slides of 12 ears with retraction pocket were histopathologically studied under microscope, especially focusing on the location of retraction pocket and inflammatory pathology in the local middle ear cavity next to retraction pockets. The temporal bone slides of 11 ears with acquired cholesteatoma were histopathologically observed and 33 cases diagnosed as acquired cholesteatoma were clinically observed observed in the local middle ear cavity next to the part without retraction pocket of eardrum. The results of pathological observation of the temporal bone slides with acquired cholesteatoma and clinical observation during operation for acquired cholesteatoma show that cholesteatoma invade mainly and occupied the ossicular chain eara of the middle ear cleft.

CONCLUSIONIn the pathological process of otitis media, the intractable pathological changes in the ossicular chain area can inward adhere posterosuperior quadrant or pars flaccida of the eardrum to form retraction pocket and permanently infiltrate the external squamous epithelial layer of retraction pocket to excessively proliferate and keratinize, leading to formation of acquired cholesteatoma.

Adolescent ; Adult ; Aged ; Aged, 80 and over ; Child ; Cholesteatoma, Middle Ear ; pathology ; Ear, Middle ; pathology ; Female ; Humans ; Inflammation ; Male ; Middle Aged ; Otitis Media ; pathology ; Tympanic Membrane Perforation ; pathology ; Young Adult

OBJECTIVETo summarize our experiences in the diagnosis and treatment of placenta accreta in the second trimester of pregnancy.

METHODSWe retrospectively analyzed the clinical data of 31 patients were admitted to Peking Union Medical College Hospital with placenta accreta in the second trimester of pregnancy from January 2002 to January 2010.

RESULTSAmong 31 cases, one case (3.2%) was suspected to be with placenta accreta by ultrasound examination and 30 cases (96.8%) were normal before delivery. Placenta accreta was identified during follow-up in 12 cases (38.7%) after delivery. Fourteen patients underwent curettage again after delivery,which was effective in 6 patients (42.9%) and failed in 8 patients,in whom uterine artery embolization (UAE) was further applied. Thirteen patients underwent UAE without curettage. In total,21 cases underwent UAE, which was effective in 19 patients (90.5%); one patient with abnormal β-human chorionic gonadotropin (β-HCG) 5 months after embolization underwent lesion resection and one case with slightly increased β-HCG were lost to follow-up. Hysteroscopy was effective in 3 patients,of whom two patients underwent lesion resection by hysteroscopy and one case who was suspected to be with trophoblastic disease by ultrasonography before surgery and confirmed to be placenta accreta during hysteroscopy examination underwent lesion resection. One case experienced hemorrhagic shock during vaginal delivery and underwent emergency laparotomy. Among all these 31 patients,massive hemorrhage occurred in 13 cases during delivery and hemorrhagic shock in 2 cases. Three cases had postpartum hemorrhage and stopped bleeding after UAE. None needed hysterectomy.

CONCLUSIONSPlacenta accreta in the second trimester of pregnancy is usually diagnosed after childbirth,which may be delayed in some cases. Therefore,special attention should be paid to this disease during follow-up. Conservative treatment was the main therapy of placenta accreta in the second trimester of pregnancy. UAE is effective in stopping bleeding.

Adult ; Dilatation and Curettage ; Female ; Follow-Up Studies ; Humans ; Placenta Accreta ; diagnosis ; therapy ; Pregnancy ; Pregnancy Trimester, Second ; Retrospective Studies ; Uterine Artery Embolization ; Young Adult

OBJECTIVETo investigate the multiple iron metabolism-related genes expression, its regulation by iron and the expression correlation among the genes in rat tissues.

METHODSTwo groups (n=30) of Sprague-Dawley female weanling rats were fed with a control diet and an iron deficient diet respectively for 4 weeks. All rats were then sacrificed, and blood and tissue samples were collected. The routine blood examination was performed with a veterinary automatic blood cell analyzer. Elemental iron levels in liver, spleen and serum were determined by atomic absorption spectrophotometry. The mRNA expression of genes was detected by real-time fluorescence quantitative PCR.

RESULTSAfter 4 weeks, the hemoglobin (Hb) level and red blood cell (RBC) count were significantly lower in the iron deficient group compared with those in the control group. The iron levels in liver, spleen and serum in the iron deficient group were significantly lower than those in the control group. In reference to small intestine, the relative expression of each iron-related gene varied in the different tissues. Under the iron deficiency, the expression of these genes changed in a tissue-specific manner. The expression of most of the genes significantly correlated in intestine, spleen and lung, but few correlated in liver, heart and kidney.

CONCLUSIONFindings from our study provides new understandings about the relative expression, regulation by iron and correlation among the mRNA expressions of transferrin receptors 1 and 2, divalent metal transporter 1, ferritin, iron regulation proteins 1 and 2, hereditary hemochromatosis protein, hepcidin, ferroportin 1 and hephaestin in intestine, liver, spleen, kidney, heart, and lung of rat.

Animals ; Ferritins ; blood ; Gene Expression ; Hepcidins ; Iron ; Liver ; metabolism ; Rats ; Rats, Sprague-Dawley

OBJECTIVETo prepare the alpha-asarone reservoir patch and investigate its release and transdermal absorption characteristics in vitro. The efficient enhancers were chosen to improve the drug's permeation rate.

METHODThe alpha-asarone reservoir patch was prepared using 1% hydroxypropyl methylcellulose (HPMC) of ethanol solution as medium and ethylene vinyl acetate (EVA) membrane to control the release of drug. The Franz diffusion cells were used and several permeation enhancers were evaluated. High performance liquid chromatorgraphy (HPLC) was used to determine alpha-asarone's content and permeation rate.

RESULTThe release mechanisms of alpha K-asarone patch in vitro coincided with zero-order kinetic. 30% ethanol cooperates with 1% Isopropyl Myristate (IPM) have the best effect on permeation of the patch. The permeation rate reaches (20.67 +/- 1.33) microg x cm(-2) h(-1).

CONCLUSIONEthanol combined with IPM is good permeation enhancer, which facilitated the permeation of alpha K-asarone to fit the clinical requirements. However, the further studies of the skin's stimulation and bioavailability are needed.

Acorus ; chemistry ; Administration, Cutaneous ; Anisoles ; administration & dosage ; isolation & purification ; pharmacokinetics ; Delayed-Action Preparations ; administration & dosage ; pharmacokinetics ; Ethanol ; pharmacology ; Humans ; Hypromellose Derivatives ; In Vitro Techniques ; Methylcellulose ; analogs & derivatives ; chemistry ; Myristates ; pharmacology ; Plants, Medicinal ; chemistry ; Polyvinyls ; chemistry ; Skin ; drug effects ; metabolism ; Skin Absorption ; drug effects

The effects of over-expression of testis-specific expressed gene 1 (TSEG-1) on the viability and apoptosis of cultured spermatogonial GC-1spg cells were investigated, and the immortal spermatogonial cell line GC-1spg (CRL-2053™) was obtained as the cell model in order to explore the function of TSEG-1. We transfected the eukaryotic vector of TSEG-1, named as pEGFP-TSEG-1 into cultured spermatogonial GC-1spg cells. Over-expression of TSEG-1 inhibited the proliferation of GC-1spg cells, and arrested cell cycle slightly at G0/G1 phase. Transfection of TSEG-1 attenuated the transcript levels of Ki-67, PCNA and cyclin D1. In addition, over-expression of TSEG-1 induced early and late apoptosis, and reduced the mitochondrial membrane potential of GC-1spg cells. Moreover, transfection of TSEG-1 significantly enhanced the ratio of Bax/Bcl-2 and transcript levels of caspase 9, and decreased the expression of Fas and caspase 8 in GC-1spg cells. These results indicated over-expression of TSEG-1 suppresses the proliferation and induces the apoptosis of GC-1spg cells, which establishes a basis for further study on the function of TSEG-1.

Objective To learn the infection status of sexually transmitted diseases (STDs)and acquired immunodeficiency syndrome (AIDS)among female sex workers (FSWs)in Huzhou,and to learn the awareness rate of knowledge of AIDS, and to provide scientific evidence for the development of prevention and control measures.Methods According to the national AIDS surveillance standards,a questionnaire survey was conducted among FSWs and blood samples were collected to detect the antibodies to human immunodeficiency virus (HIV),syphilis and hepatitis C virus.Results A total of 800 FSWs,with an average age (25. 5 ±5. 0 years old)were investigated,and the awareness rate of AIDS related knowledge was 92. 0%.The positive rate of syphilis antibody was 2. 9%and the positive rate of hepatitis C antibody was 0. 2%.HIV antibody was tested negative.The persistent condom use rate among commercial sex workers was 57. 6% in the recent month.The awareness rate of AIDs related knowledge among FSWs with senior middle school education and above was 3. 51 times of those with primary school education and below.Conclusion The prevalence of syphilis and hepatitis C infection among FSWs were high in Huzhou.It is necessary to strengthen the health education among FSWs to improve the awareness of the knowledge,especially for those with low level education.And it is necessary to improve the accessibility of AIDs prevention service to reduce the spread of HIV.