Objective To evaluate the efficacy and safety of a levonorgestrel-releasing intrauterine system(LNG-IUS)for the treatment of dysmenorrhea associated with adenomyosis.Methods We recruited 48 women with moderate or severe dysmenorrhea associated with adenomyosis.All women were inserted of LNG-IUS into their uterine cavity from days 5-7 of their periods and maintained for 12 months.We compared the visual analogue scale(VAS)scores and verbal rating scale(VRS)scores of their dysmenorrhea and dyspareunia at baseline and 12 monthes follow-up.Results Forty-four women completed the study. There were significant differences between mean VAS and VRS scores changes of dysmenorrhea and dyspareunia at baseline and 12 monthes follow-up,those of dysmenorrhea dropping from 75?13 to 11?11 and 2.3?0.4 to 0.4?0.3,those of dyspareunia dropping from 54?19 to 4?4 and from 1.6?0.8 to 0.2?0.2 respectively.Overall 29 women(66%)were very satisfied or satisfied with the one-year treatment. Conclusion Insertion of LNG-IUS alleviates moderate or severe dysmenorrhea associated with adenomyosis remarkably.

Objective To investigate new type cytokine induced killer cells expansion using advanced breast cancer′s peripheral blood .Methods peripheral blood mononuclear cells were isolated from 8 advanced breast cancer volunteers and co -cultured with Cytokine induced killer cells .These cells were placed in plastic flasks containing CIK-MediumTM supplemented with 10% auto-plasma in the presence of IL -2 ( 1 000 IU/mL) .The cultures were fed with CIK-MediumTM supplemented with IL -2 following the proliferation capacity . Cell proliferation was measured by cell counting during the cultivation .Fourteen days after cultivation ,cell mark-ers CD3/CD16/CD56 were examined by flow cytometry .51Cr and MTT assays were employed in cytotoxicity as-says.Cytokines were assayed by ELISA method .Results CD16+,CD16+CD56+,CD56+CIK cells were 5.8~11.6%in 2 ×107 fresh PBMCs and 95.2~97.6%in co-cultured cells after 18 days cultivation .The in vitro ex-pansion rate of new type cytokine induced killer cells was up to more than 8.2 ×108 in total,the cytotoxicity are ef-fective killing cells against MCF 7 and BT20 breast cancer cell lines .New type cytokine induced killer cells expand-ed from all PBMCs and secreted cytokines IFN -and TNF-.Conclusion The present culture could be useful to clarify the mechanisms of CIK cells expansion in vitro and feasible for breast cancer immmuno cell therapy .

OBJECTIVEPrader-Willi syndrome (PWS) is an example of a human genetic disorder that involves imprinting genes on the proximal long arm of chromosome 15 and SNRPN gene as a candidate gene for this syndrome. The purpose of this study was to show the molecular genetic defects and genomic imprinting basis in Chinese PWS patients and to evaluate the clinical applications of a differential diagnostic test for PWS.

METHODSFluorescence in situ hybridization (FISH) and methylation-specific PCR (MSPCR) techniques were applied for 4 clinically suspected PWS patients. Using three probes, including SNRPN probe for identification of the critical locus in PWS region, D15Z1 and PML control probes for identification of the 15p arm and 15q arm, the authors detected the deletions 15q in PWS. MSPCR was based on sodium bisulfite treatment of DNA and PCR primers specific for the maternal and paternal allele.

RESULTSWhen hybridized with mixed probes, it was found in 2 patients that the central specific signal was absent, but both the flanking control signals were retained, indicating SNRPN gene deletion of chromosome 15q11-13. Bisulfite-modified DNA from all PWS children amplified with methylated allele-specific primer pair showed only maternal 131bp PCR product, indicating the maternal uniparental disomy (UPD15).

CONCLUSIONGenomic imprinting plays an important role in the molecular pathogenesis of PWS that caused by paternal microdeletions of 15q11-q13 or maternal UPD of chromosome 15. The basic defect seemed to be an absence of function of PWS genes that are normally expressed only from the paternal chromosome 15. MSPCR is a rapid and simple PCR-based assay compared with other cyto-molecular tests and its results were consistent with the clinical diagnosis of PWS, so it seems to be a reliable diagnostic method for PWS patients who show abnormal methylation at SNRPN. The genetic differential tests for PWS are important in determining familial recurrence risk.

Adolescent ; Autoantigens ; Chromosome Deletion ; Chromosomes, Human, Pair 15 ; genetics ; Gene Deletion ; Genomic Imprinting ; genetics ; Humans ; In Situ Hybridization, Fluorescence ; Male ; Polymerase Chain Reaction ; methods ; Prader-Willi Syndrome ; genetics ; Ribonucleoproteins, Small Nuclear ; genetics ; snRNP Core Proteins

The transgenic mice that express Cre recombinase in a tissue specific manner is a powerful tool in generating the conditional gene knockout mice. The rat insulin promoter was cloned target the expression of Cre in pancreatic tissue. The Cre gene was modified by adding the nuclear localization signal and the sequence for initiation by eukaryotic ribosomes at 5' terminal of the Cre gene. Cre gene was linked to the intron of human growth factor gene. This construct was introduced into the mouse eggs using microinjection. Seven mice were identified as founders carrying the Cre gene by PCR. The results of RT-PCR showed that the transgenic mouse from one founder could transcribe the foreign gene in pancreas. The Southern blot analysis indicated that the Cre recombinase expressed in pancreas of the transgenic mouse was functional.
Animals ; Blotting, Southern ; Female ; Insulin ; genetics ; Integrases ; genetics ; Mice ; Mice, Transgenic ; Pancreas ; metabolism ; Promoter Regions, Genetic ; RNA, Messenger ; analysis ; Rats ; Reverse Transcriptase Polymerase Chain Reaction ; Viral Proteins ; genetics

Objective:To observe the effects of different intervention schemes on the expression of adhesion-related cytokines, menstrual recovery and clinical outcome of patients after transcervical resection of adhesion (TCRA) .Methods:180 patients received TCRA in our hospital from Feb. 2022 to Feb. 2023 were divided into group A, group B and group C according to different post-operative intervention programs, with 60 patients in each group. Patients in group A were treated with artificial cycle of estrogen and progesterone after surgery. On this basis, patients in group B were placed with a uterine birth control ring, and patients in group C were injected with sodium hyaluronate gel into the uterine cavity. The grade of uterine cavity adhesion, improvement rate of menstruation and pregnancy outcome at 2 months after operation and pregnancy outcome within 1 year after surgery were compared between the three groups at 2 months after operation. The relative mRNA expression of endometrial tissue transforming growth factor β1 (TGF-β1), platelet-derived growth factor BB (PDGF-BB), tissue inhibitor of matrix metalloproteinase-1 (TIMP-1) and basic fibroblast growth factor (bFGF) at uterine cavity adhesion in each group were detected and compared.Results:At 2 months after surgery, the uterine adhesion rates in group A, group B, and group C were 43.33%, 15.00%, and 11.67%, respectively. There was no significant difference in the uterine adhesion rates between group B and group C ( P>0.05), but they were significantly lower than those in group A ( P<0.05) ; Meanwhile, the degree of intrauterine adhesions in group B and group C was significantly milder than that in group A ( P<0.05). The menstrual improvement rates of group A, group B, and group C at 2 months after surgery were 76.67%, 93.33%, and 96.67%, respectively. There was no significant difference between group B and group C ( P>0.05), but they were all significantly higher than group A ( P<0.05). At 2 months post surgery, the relative expression levels of TGF-β1, PDGF-BB, TIMP-1, and bFGF mRNA in the endometrial tissue at the site of uterine adhesions in group A were 0.77±0.26, 0.58±0.27, 0.54±0.15, and 0.62±0.14, respectively. In group B, they were 0.37±0.16, 0.37±0.14, 0.26±0.11, and 0.29±0.10, respectively. In group C, they were 0.32±0.16, 0.21±0.09, 0.27±0.08, and 0.34±0.18, respectively. The relative expression levels of cytokines in each group were significantly lower than during surgery ( P<0.05). There was no significant difference in the relative expression levels of various cytokines mRNA between group B and group C at 2 months after surgery ( P>0.05), but both were significantly lower than group A ( P<0.05). The pregnancy success rates within 1 year after surgery in group A, group B, and group C were 40.00%, 55.00%, and 58.33%, respectively. The pregnancy success rate in group C was significantly higher than that in group A ( P<0.05) . Conclusions:The application of metauterine contraceptive ring or sodium hyaluronate gel on the basis of estrogen and progesterone treatment after TCRA can effectively prevent postoperative re-adhesion of patients with intrauterine adhesions, improve clinical symptoms, and reduce the expression level of adhesion cytokines. The effects of the two schemes are equivalent.

OBJECTIVETo explore the risk factors of acute lymphoblastic leukemia (ALL) recurrence in adult patients and establish a prognosis index (PI) calculation model in order to improve the prevention strategy of ALL in adults.

METHODS104 adult ALL patients from Blood Diseases Hospital & Chinese Academy of Medical Sciences between August 2008 and November 2011 were enrolled. COX proportional hazards regression stratified by Dummy variable was used to set up the prediction model; Kaplan-Meier method and Log-rank test were used to estimate and compare the survival. After calculated individual PI value, patients' expected survival should be estimated by groups.

RESULTSThe overall median survival of adult ALL patients was 22.00 months (95% CI 17.00-27.00). COX regression analysis showed that chemotherapy group patients had a higher risk of recurrence than of ASCT group while setting treatment as the dummy variable (RR=2.052, 95%CI 0.877-4.799, P=0.007). Stratified Analysis showed that the risk factors of B-ALL recurrence in adult patients included HGB <100 g/L (RR=0.186, 95% CI 0.068-0.512, P=0.001), CNSL (RR=7.767,95% CI 2.951- 20.433, P=0.001), number of consolidation chemotherapy<3 (RR=0.445, 95% CI 0.211-0.940, P=0.034) and Ph chromosome positive (RR=2.771, 95% CI 1.353-5.674, P=0.005). Grouped by the PI value, the expected survival of each individual patient could be estimated as PI=0.58 base.

CONCLUSIONHGB, CNSL, number of consolidation chemotherapy and Ph chromosome were independent risk factors of B-ALL recurrence in adult patients. PI value could predict the survival of adult ALL patients and provide reference for individual therapy and prognostic evaluation.

Adolescent ; Adult ; Female ; Humans ; Male ; Middle Aged ; Precursor Cell Lymphoblastic Leukemia-Lymphoma ; epidemiology ; pathology ; Prognosis ; Recurrence ; Risk Assessment ; Risk Factors ; Young Adult

Objective To summarize the treatment principle of acupuncture-moxibustion in treating Bi-impediment syndrome from the application rules of meridians and acupoints in Ming-Qing Dynasties by sorting out and analyzing the Chinese medicine literatures about acupuncture-moxibustion for Bi-impediment syndrome in Ming-Qing Dynasties, for providing literature evidence for basic and clinical research of Bi-impediment syndrome.Method Via electronic retrieval ofZhong Hua Yi Dian (Zhen Jiu Tui Na Lei) (Chinese Medical Encyclopedia,Chapter of Acupuncture-Moxibustion and Tuina), the data related to Bi-impediment syndrome in Ming-Qing Dynasties were extracted to establish a database categorized by meridians and acupoint features in Excel for analysis.Result There were 267 items of records about acupuncture-moxibustion in treating Bi-impediment syndrome in Ming-Qing Dynasties, involving the fourteen ordinary meridians, and 131 acupoints including 5 extra points; the frequency of using the Gallbladder Meridian ranked the top, followed by the Large Intestine Meridian; points from the Bladder Meridian were predominant, followed by the Gallbladder Meridian; there were 28 commonly-used acupoints (frequency>5), which were Quchi (LI 11, 26 times), Huantiao (GB 30, 23 times), Hegu (LI 4, 22 times), Chize (LU 5, 16 times),Yanglingquan (GB 34, 15 times), and Weizhong (BL 40, 14 times). Of the specific acupoint, the five Shu points were most frequently used, with a frequency of 217.Conclusion In the treatment of Bi-impediment syndrome with acupuncture-moxibustion, doctors in Ming and Qing Dynasties selected yang meridians more often than yin meridians, and Gallbladder, Large Intestine and Bladder Meridians had comparatively higher frequencies; regarding the application of acupoints, the specific acupoints were often used, especially the five Shu acupoints. The study results provide reference for acupoint selection in the treatment of Bi-impediment syndrome with acupuncture-moxibustion.

OBJECTIVETo investigate the expression of PDGF receptor-beta and its correlation with extracellular matrix in hepatic tissue during hepatic fibrosis.

METHODSThe model of hepatic fibrosis in rats was induced by carbon tetrachloride. PDGF receptor-beta subunit, collagen I, collagen III and a-SMA in hepatic tissues of these rats were examined using immunohistochemistry. The correlation between PDGF receptor-beta subunit and collagen I, III was analyzed using SAS software after the results of immunohistochemistry were semi-quantified.

RESULTSPDGF receptor-beta subunit and a-SMA were not detected in normal controls. Collagen I and III were distributed in the portal tracts and beneath the endothelia of the central veins and of the Disse spaces. Two weeks after CCl4 injection, the PDGF receptor-beta and a-SMA were detected, and the expression of collagen I and III increased. At the end of 4 and 6 weeks, the above four proteins were further increased. Two weeks after CCl4 injection, PDGF receptor-beta had no apparent correlation with collagen I and III. However, PDGF receptor-beta had a significant correlation with collagen I and III 2 weeks later, and the correlation coefficient was 0.74 and 0.60 respectively at 4 weeks, and 0.83 and 0.67 respectively at 6 weeks. PDGF receptor-beta had a significant correlation with a-SMA during the whole process of hepatic fibrosis and the correlation coefficient was 0.62, 0.69 and 0.81, respectively at the time of 2, 4 and 6 weeks after CCl4 injection.

CONCLUSIONThe PDGF receptor-beta was overexpressed during the process of hepatic fibrosis development, and it significantly correlated with collagen I and collagen III.

Animals ; Carbon Tetrachloride ; Carbon Tetrachloride Poisoning ; Collagen Type I ; biosynthesis ; genetics ; Collagen Type III ; biosynthesis ; genetics ; Extracellular Matrix ; metabolism ; Liver ; metabolism ; Liver Cirrhosis, Experimental ; chemically induced ; metabolism ; Male ; Rats ; Rats, Sprague-Dawley ; Receptor, Platelet-Derived Growth Factor beta ; biosynthesis ; genetics

OBJECTIVETo study the cleavage activity of hammerhead ribozyme targeting at platelet-derived growth factor receptor beta subunit (PDGFR- beta) mRNA in hepatic stellate cells (HSCs) and its effect on the biological characters of HSCs.

METHODSExpression vector of anti-PDGFR- beta ribozyme was constructed and transfected into rat-derived HSC-T6 cells with lipofectin. The positive cell clones were gained by G418 selection. The expression of PDGFR- beta, alpha-smooth muscle actin (alpha-SMA), and type I and type III collagen was detected by means of northern blot, Western blot and immunocytochemical staining respectively. The cell proliferation was determined with MTT colorimetric assay. The cell apoptosis was demonstrated with flow cytometry, acridine orange fluorescence vital staining and transmission electron microscopy.

RESULTSThe expression of PDGFR- beta at mRNA and protein level was markedly reduced in ribozyme-transfected HSCs only 43% to 51% of that in control cells (t > or = 3.957, P < 0.05), and alpha-SMA expression level, type I and type III collagen synthesis ability were also reduced (t > or = 6.790, P < 0.01). The proliferation of ribozyme-transfected HSCs was significantly decreased (t > or = 3.858, P < 0.05), and the proliferation response to PDGF BB was markedly inhibited. However the apoptotic rate was significantly increased in ribozyme-transfected HSCs (chi2 > or = 14.157, P < 0.01), and typical apoptotic cells could be found under transmission electron microscopy.

CONCLUSIONSThe anti-PDGFR- beta ribozyme can be expressed stably in HSCs, cleave the target RNA effectively, inhibit HSCs proliferation and collagen synthesis, and induce HSC apoptosis. The results suggest that inhibiting PDGFR- beta expression in HSCs may be a new therapy for liver fibrosis.

Apoptosis ; drug effects ; Cell Division ; Cells, Cultured ; Hepatocytes ; drug effects ; physiology ; Humans ; Liver ; pathology ; RNA, Catalytic ; pharmacology ; RNA, Messenger ; biosynthesis ; Receptor, Platelet-Derived Growth Factor beta ; antagonists & inhibitors ; metabolism

OBJECTIVETo investigate the features of HBx protein distributed in liver cells and its expression in E. coli.

METHODSThe expression vectors encoding the full length HBx and its mutants were constructed by the routine molecular cloning method. HBx protein expression was detected using Western blotting. The distribution feature of HBx protein in liver cells was examined using the fluorescence confocal microscopy. A series of purified HBx fusion proteins were obtained by glutathione-sepharose 4B affinity chromatography.

RESULTSThe expression vectors were successfully constructed for the full length HBx and its mutants. HBx was found distributed uniformly in the nuclei but granularly in the cytoplasm of the liver cells. Under optimal conditions, the mutant GST-HBx (72-120aa) was easily degraded.

CONCLUSIONThis study may provide a basis for further study on the biological function of HBx at the protein level.

Carcinoma, Hepatocellular ; pathology ; Cell Line ; Cloning, Molecular ; Escherichia coli ; metabolism ; Genetic Vectors ; Glutathione Transferase ; biosynthesis ; genetics ; Hepatocytes ; cytology ; metabolism ; Humans ; Liver ; cytology ; Liver Neoplasms ; pathology ; Mutation ; Recombinant Fusion Proteins ; biosynthesis ; genetics ; Trans-Activators ; biosynthesis ; genetics ; Tumor Cells, Cultured