ObjectiveTo study microRNA (miRNA) expression and role of cell cycle regulation in decidualized endometrial stormal cells (ESC) in vitro.MethodsESC was induced decasualization in vitro and matched with non-decidualized cells as controls.The expression repertoire of miRNA was measured by microarray chip and was validated by real-time PCR.Flow cytometry was used to identify ESC cycle during decidual reaction in vitro and after miRNA222 inhibitor was transfected into it.Results (1) Between decidualized and undecidualized stromal cells,there were 49 miRNAs significantly different expression by microarray chip,including 16 miRNA up-regulation and 33 miRNA down-regulation.hsa-miR-27b,30c,143,101,181 b,29b,30d,507,23 a,222,221 exhibited significantly differential expression between decicualized and undecidualized stromal cells by real-time PCR (P ＜0.05).(2) After miRNA222 inhibitor (NC-FAM) transfection to decidual ESC,ESC were cultured by FBS medium for 24 hours,the rate of transfection was 70％.ESC were transfected with miRNA 222 inhibitor and cultured for 48 hours,the percentage of ESC at Sphase of (6.2 ± 0.7 ) ％ were significantly lower than ( 10.9 ± 0.8 ) ％ in control group ( P ＜ 0.05 ) ; the percentage of ESC at G0/G1 phase increased at transfection group [ (77.5 ± 1.3 ) ％ vs.(73.0 ± 1.6) ％ at control group ],but there was no significant difference (P ＞ 0.05 ).Decasualization ESC were transfected with miRNA 222 inhibitor and cultured for 48 h,the percentage of ESC at S-phase was ( 3.3 ± 0.6) ％ in transfection group,which were significantly lower than (7.8 ± 0.9 ) ％ in control group ( P ＜ 0.05 ).The percentage of ESC at G0/G1 phase was ( 80.7 ± 1.6 ) ％ in transfection group and ( 74.9 ± 1.1 ) ％.In control group,which did not reached statistical difference ( P ＞ 0.05).ConclusionmiRNA was involved in ESC decidual process in vitro by regulating cell cycle.

Aim To observe the effects of oral puerarin (Pue) on the maternal gestational diabetes mellitus (GDM) rats and its fetal growth and development, so as to provide references for the application of Pue in the treatment of GDM. Methods The GDM rat model was established by injecting streptozotocin (STZ) into the tail vein of pregnant female rats, and the rats were treated with Pue orally for 12 days; the body weight and abortion of pregnant rats were recorded. The fasting blood glucose of pregnant rats was detected before and at the end of the treatment, and the glucose tolerance was tested on the 5th and 10th days after the administration of the drug. The cesarean sections were carried out on the 20th day of pregnancy. The blood glucose content of the fetal rats was detected, and the state of development was observed. The body weight and length were measured, as well as the placenta and the important organs weight, and the indexes of the organs were calculated. Results Compared with the model group, Pue could significantly reduce the fasting blood glucose of GDM pregnant rats and fetal rats, improve the glucose tolerance of pregnant rats, effectively alleviate the excessive weight gain of pregnant rats and overweight of fetal rats caused by GDM, and reduce the abortion rate; it could also reverse the decrease in the indexes of the organs of brain, heart, and liver, and the increase in the indexes of organs of kidney in fetal rats caused by GDM. Conclusions Pue can relieve the maternal and the fetal hyperglycemia in GDM, reduce the rate of miscarriage, reduce the incidence of macrosomia, and promote the development of vital fetal organs.

Objective To investigate the effect of small interfering RNA (siRNA) mediated silencing of △Np73 on 5-FU chemotherapy sensitivity in SW620 colocancer cells and provide new treatment approach for the colon cancer.Methods siRNAs were transfected into SW620 colon cancer cells.The expression of △Np73 was observed.Cell viability of colon cancer cells were measured by MTr assay and cell apoptosis was assessed with flow cytometry after treatment of control siRNA or △Np73 siRNA or combined with 5-FU,respectively.The tumorigenesis was assessed by injecting △Np73 siRNA or control siRNA transfeeted SW620 colon cancer cells into nude mice,followed by treatment with 5-FU in the tumors.Results △Np73 siRNA was able to strongly inhibit △Np73 expression,however,it did not inhibit the growth of cells.Combination treatment with △Np73 siRNA and 5-FU produced significant higher apoptotic cell(42.9%) as compared with those with 5-FU treatment(18.9%) alone or those with △Np73 siRNA(8.8%) alone.The treatment with 5-FU in the xenngrafts derived from △Np73 siRNA transfected SW620 cells in nude mice can inhibitor tumor growth significantly (t=15.32,P<0.05).Conclusion △Np73siRNAs can specifically repress the expression of △Np73.Thus it sensitizess the cells to 5-FU chemotherapy in colon cancer.

Objective To study the immunosuppressive mechanism of alkaloid sinomenine (SIN) by observing the effects of SIN on the proliferation and intracellular protein expression levels of nuclear factor of activated T cells (NF-AT) and interferon-gamma (IFN-γ) in CD4+ T lymphocytes of human periphery blood. Methods CD4+ T lymphocytes were isolated from PBMC suspensions with immunomagnetic beads and divided into five groups to culture. (1) Negative control group: no medicine was added to cell culture medium; (2) Positive control group: CsA solution (final concentration: 50ng/ml) was added to cell culture media; (3) Low-concentration SIN group (L-SIN): low-concentration SIN solution (final concentration: 10 μmol/L) was added to cell culture media; (4) Middle-concentration SIN group (M-SIN): middle-concentration SIN solution (final concentration: 200 μmol/L) was added to cell culture media; (5) High-concentration SIN group (H-SIN): high-concentration SIN solution (final concentration: 1000 tmol/L) was added to cell culture media. The proliferations of CD4+ T lymphocytes were observed. Western blotting was performed to detect the protein expression levels of NF-AT. FCM was used to determine the levels of IFN-γ. Results Compared with negative control group, the cell proliferation was significantly inhibited in H- and M-SIN groups (P＜0. 01 ). SIN concentration-dependently inhibited the protein expression levels of NF-AT and IFN-γ in CD4+ T lymphocytes of human periphery blood (P＜0.01). The protein expression levels of NF-AT and IFN-γ were lowest in positive control group. There was a close negative correlation between intracellular levels of NF-AT and cell proliferation inhibition ratio in CD4+ T lymphocytes of human periphery blood (rs = - 0. 969, P = 0. 000). Conclusion SIN can inhibit the protein expression of NF-AT and IFN-γ in CD4+ T lymphocytes of human periphery blood probably by decreasing protein levels of NF-AT to inhibit the activity and proliferation of CD4+ T lymphocytes.

Objective To analyze the epidemiologic characteristics and risk factors for mortality in non-(human immunodeficiency virus,HIV) infected children with pneumocystis carinii pneumonia(PCP). Methods The data of non-HIV infected children with PCP diagnosed in Beijing Children′s Hospital from January 1,2006 to December 31,2012 were collected. They were divided into survival and non-survival group according to the prognosis. The epidemiologic characteristics and risk factors for mortality were analyzed. Results Sixteen patients were enrolled in this study. Ten of them survived and 6 of them were non-survived. The basic diseases included malignant tumor in 5 patients and non-malignancy diseases in 11 of them. Com-pared with the survival group,the non-survival group had a higher average age [(12. 00 ± 2. 00) years vs. (6. 65 ± 4. 32)years,P=0. 01],higher ratio to need mechanical ventilation (6/6 vs. 4/10,P=0. 04),lower PaO2/FiO2[(73. 88 ±26. 95) mmHg vs. (167. 50 ± 97. 17) mmHg,1 mmHg=0. 133 kPa,P=0. 01] and lower pediatric critical illness score(75. 67 ± 5. 72 vs. 86. 40 ± 8. 88,P=0. 02). There were no differences on sex ratio,kinds of basic diseases,whether with co-infections,the time of immunosuppressant administration, the time from onset to diagnosis,the time from onset to beginning trimethoprim-sulfamethoxazole therapy, PaCO2 ,white blood cell counts,lymphocyte counts,CD4+ cell counts,C-reactive protein,and hemoglobin con-centrations between the survival and non-survival group. Conclusion A higher age, need for mechanical ventilation,lower PaO2/FiO2 and lower pediatric critical illness score were risk factors for mortality in non-HIV infected children with PCP.

Objective To discuss the clinical application of ultrasound-CT double-guided radiofre-quency ablation(RFA) in treating hepatic tumors. Methods Fifteen patients with 20 liver tumor lesions were included in this study. Ultrasound-CT double-guided radiofrequency ablation was employed in all patients. First, under ultrasound guidance the electrode of RFA was inserted to the site close to the lesion, then, guided by CT scanning the accurate positioning of the electrode was accomplished and RFA procedure was completed. Results The accurate puncturing of the electrode was achieved in all 20 hepatic lesions, and the RFA procedure was successfully performed in a short time. Follow-up examination showed that there was no obvious residual tumor tissue, and no RFA-related complications occurred in all the 15 patients. Conclusion Under ultrasound-CT double-guidance, the percutaneous transhepatic puncturing can be more accurately accomplished, which can ensure a successful RFA procedure. This technique is especially useful when the hepatic tumor is incomplete or unclear on ultrasonograph, and it can improve the puncturing accuracy and reduce the complications as well.

Objective To analyze laparoscopic lymphadenectomy and the safety of gastrectomy for gastroesophageal junction cancer.Methods From Jan 2011 to Dec 2012 72 gastroesophageal junction cancer patients were enrolled, including 46 patients in laparoscopic group, and 26 in open surgery group.Results There was no significant difference in the numgbers of lymph node dissection between the two groups and nor difference in the number of positive lymph node dissection.Esophagus resection length in open group was (2.0 ± 1.0) cm, while that was (3.0 ± 0.8) cm in laparoscopic group (t =0.471, P ＜ 0.001).5 (19％)patients in open group had positive margins compared to six in laparoscopic group (13％), x2 =0.491, P =0.483.7 patients in the open group underwent thoracoabdominal resection, while in laparoscopic group 3 patients did, x2 =5.781, P =0.016.Laparoscopic splenic hilar lymphnodes dissection harvested more lymph nodes (t =0.260, P =0.011).Laparoscopic gastrectomy used less operation time (t =0.237, P =0.021) experinced less blood loss (t =0.451, P ＜ 0.01) than open group.There was no difference in major complications between the two groups.Conclusions Laparoscopic splenic hilar lymphnodes dissection in gastroesophageal junction cancer surgery is superior to open surgery, with more lymph nodes harvested, longer esophageal cutting distances, lower incidence of thoracoabdominal surgery, shorter operation time, and less blood loss.

Objective:To observe the clinical function of Chinese medicine enema liquid combined with antibiotics in syndrome differentiation typing treatment of chronic pelvic inflammation. Methods:A total of 240 patients with chronic pelvic inflammation were randomly divided into the control group(120)and the observation groups(120)including damp-heat stasis accumulation type,qi stagnation and blood stasis type and cold-dampness accumulation type. The patients in the control group received 0. 2g levofloxacin hydrochloride with intravenous drip,twice a day for 14 days,while those in the observation groups were given 250ml Chinese medicine enteroelysis(damp-heat stasis accumulation type was given Baiyi enteroelysis,qi stagnation and blood stasis type was given Puyao enteroelysis,and cold-dampness accumulation type was given Baiyao enteroelysis )with irrigation treatment once a day additionally. The patients in the observation group were given enteroelysis three or five days after menstruation and continued for 14 days,and the treatment course was two menstrual cycles. The indices of hemorheology and clinical efficacy were compared after the treatment between the groups. Results:The total effective rate in the observation group (91. 7% )was higher than that in the control group(81. 7% )(P < 0. 05). The improvement of whole blood viscosity,plasma viscosity,fibrinogen,erythrocyte sedimentation rate and platelet aggregation rate in the observation group were better than those in the control group(P < 0. 05). Conclusion:Chinese medicine enteroelysis shows obvious curative effect in the treatment of chronic pelvic inflammatory diseases,and combined with antibiotics,the effect can be strengthened,which is worthy of clinical application.

Objective To estimate the effect of a tretinoin derivative ECPIRM on retinoic acid receptors (RARs),and to observe skin irritation responses to it in mice.Methods Cultured SCL-1 cells were divided into 2 groups to be treated with culture medium containing 10 μmol/L ECPIRM (ECPIRM group) or 10 μmol/L all-trans retinoic acid (ATRA) (ATRA group) for 24 hours,and those treated with drug-free culture medium served as the control group.Western blot analysis and real-time fluorescence-based quantitative PCR were performed to quantify the protein and mRNA expressions of RARs (RARα,RARβ,RARγ and RXRα) respectively.In addition,real-time fluorescence-based quantitative PCR was conducted to measure the mRNA expressions of two target genes of the activated RAR signaling pathway,i.e.,cytochrome P450 26A1 (CYP26A1) and tazarotene-induced gene 1 (TIG1).Eight BALB/c mice were equally divided into 2 groups to be topically treated with 0.075％ ECPIRM gel or 0.05％ ATRA cream at equal molar concentrations on the shaved skin once daily for 21 successive days.Skin irritation reactions were assessed in these mice.Results Compared with the control group,the ATRA group showed significantly increased protein and mRNA expressions of RARα,RARβ and RARγ (all P ＜ 0.05).The mRNA expressions of CYP26A1 and TIG1 genes in the ATRA group were 25.49 and 3.88 times that in the control group respectively (both P ＜ 0.01).However,there was no significant difference in the protein expressions of RARα,RARβ,RARγ and RXRα,or mRNA expressions of RARα,RARβ,RARγ CYP26A1 and TIG1 between the ECPIRM group and control group (all P ＞ 0.05).Obvious Skin irritation reactions such as erythema and desquamation were observed in BALB/c mice after 2-day topical treatment with ATRA cream,and their degree peaked after 5-day treatment.However,neither erythema nor desquamation was observed in BALB/c mice during 21-day treatment with 0.075％ ECPIRM gel.Conclusion Unlike ATRA,ECPIRM cannot activate the canonical RAR signaling pathway or cause skin irritation reactions.

Objective:To evaluate arthrography-assisted reduction in minimally invasive treatment of tibial plateau fractures.Methods:From January to May 2019, 9 patients with tibial plateau fracture were treated by arthrography-assisted minimally invasive surgery at Department of Orthopaedics and Traumatology, Hong-Hui Hospital. They were 6 males and 3 females with an average age of 45.5 years (from 27 to 63 years). According to the Schatzker classification, there were 6 cases of type Ⅱ, one of type Ⅲ and 2 cases of type Ⅳ. Arthrography was used to determine the lowest filling point of contrast medium after intra-articular injection and to ensure satisfactory reduction of articular surface after fracture reduction and bone grafting, followed by minimally invasive percutaneous plate implantation. Comparisons were made between preoperation and 6 months postoperation in the range of knee motion and knee scores of The Hospital for Special Surgery (HSS). Knee X-ray films were evaluated by the Rasmussen imaging system at 6 months postoperation. Collapse of articular surface was recorded pre- and post-operation as well as postoperative complications.Results:All the 9 patients were followed up for 6 to 9 months (average, 7 months). All patients achieved bony union within 12 weeks postoperation. The preoperative knee flexion (60.7°±13.1°) and HSS (51.9±5.7) were significantly improved to 122.4°±10.8° and 84.9±5.3 at 6 months postoperation ( P<0.05), but there was no significant difference between preoperation and 6 months postoperation in knee extension (4.4°±2.5° versus 4.6°±2.9°) ( P>0.05). The collapse of articular surface was improved from preoperative 9.5 mm (from 5 to 15 mm) to postoperative 1.3 mm (from 0 to 3 mm). The Rasmussen imaging scores at 6 months postoperation showed 6 excellent and 3 good. No such postoperative complications occurred as incision infection, joint stiffness, loosening or breakage of implants. Conclusion:Arthrography helps minimally invasive treatment of tibial plateau fracture because it can better display fracture collapse during operation and accurately judge indirect reduction of articular surface.