Objective:Human capillary lymphatic endothelial cells (LECs) were isolated and cultured to assist further investigation of the function of lymphatic vessels generation during cancer metastasis. Methods:Human skin was digested by type I collagenase. Cells were isolated using magnetic beads which were marked by monoclonal antibodies against the extracellular domain of VEGFR-3, and purified by cloning col-umn. The morphology and structure of cells were observed by microscopy. Immunophenotype was identified by immunofluorescence. Cell growth curve was recorded to measure the effect of VEGF-C protein. Results:LECs exhibited the typical cobblestone morphology as monolayer growth pattem under microscopy. Enlarged nucleus and rich cytoplasm with bubbles were found under electromicroscopy. LECs specific markers inclu-ding VEGFR-3, LYVE-1, Podoplaninand D2-40 all were positive. VIII factor as specific marker of blood ves-sel endothelium cells (BVECs) was negative. VEGF-C induced a marked increase of cell proliferation. Con-clusions:Human dermal LECs could be harvested successfully using collagenase digestion, immunomagnet-ic beads sorting and clonic column purification.

Objective To investigate the clinical features ,diagnosis and treatment of cesarean scar pregnancy (CSP) .Methods We retrospectively analyzed the medical history ,clinical manifestation ,diagnoses ,treatments and outcomes of 42 patients with ec‐topic pregnancy in the cesarean scar .Results All cases got diagnosed by transvaginal ultrasound .The error rate of first diagnosis was 40 .4% (17/42) .There were 25 cases of conservative therapy ,in which 12 cases were resolved with laparoscopic surgery and 1 case resolved with open surgery ;in the first process 15 cases were carried out laparoscopic surger in which 1 case were converted to laparotomy ;abdominal surgery were underwented in 1 case and were cured .Only one case underwent abdominal total hysterectomy as of failure after uterine artery embolization .After initial conservative treatment 3 cases were hospitalized again and 2 cases under‐went reoperation .41 patients were successfully retained the uterus and normal menstrual cycle returned at two months after opera‐tion .Conclusion The early diagnosis for CSP mainly depended on ultrasound and the thickness of incision muscle layer is an impor‐tant basis for choice of treatment .Checking the thickness of incision muscle layer for CSP mainly depended on ultrasound ,and lapa‐roscopic surgery is an ideal method for diagnosis and treatment of CSP .

Objective To evaluate the effect of hydrogen sulfide combined with mild hypothermia on cerebral ischemia-reperfusion (I/R) injury in rats. Methods Eighty male SD rats, aged 3 months, weighing 250-300 g, were randomly divided into 5 groups ( n = 16 each): sham operation group (group S), cerebral I/R group,mild hypothermia group (group M), sodium hydrosulfide group (group NaHS) and NaHS + mild hypothermia group (group NM). In group I/R, M, NaHS and NM, cerebral I/R was induced by occlusion of 4 vessels (cauterization of bilateral vertebral arteries and 15 min occlusion of bilateral common carotid arteries) followed by reperfusion. In group NaHS and NM, intraperitoneal NaHS 14 μmol/kg was injected immediately after reperfusion, while the equal volume of normal saline was injected in the other three groups. At the same time, the rectal temperature was reduced to 32-33 ℃ within 15 min, lasting for 6 h, in group M and NM, while it was maintained at 36-37 ℃by physical method in other groups. Twelve rats of each group were sacrificed after 6 h of reperfusion, and then the hippocampus was removed for determination of the content of H2 S by using spectrophotometer and the expression of p-CREB and BDNF mRNA by using Western blot and RT-PCR respectively. Four rats in each group were sacririced after 72 h of reperfusion and then the hippocampus was removed for microscopic examination. Results The cerebral I/R injury was attenuated in group M, NaHS and NM compared with group I/R, with the slightest injury in group NM. The H2S content was significantly higher in group I/R, M, NaHS and NM than in group S, and in group NaHS and NM than in group I/R and M. The expression of p-CREB and BNDF mRNA was significantly higher in group I/R, M, NaHS and NM than in group S, and in group M, NaHS and NM than in group I/R. The BDNF mRNA expression was significantly higher in group NM than in group M and NaHS. There was no significant difference in the H2S content and the expression of p-CREB and BNDF mRNA between group NaHS and M.Conclusion Hydrogen sulfide combined with mild hypothermia can attenuate cerebral I/R injury by up-regulating the expression of p-CREB and BDNF mRNA in hippocampus in rats.

Objective To explore the protective effect of recombinant superoxide dismutase(SOD)fusion protein against the infection of Schistosoma japonicum Chinese strain.Methods The recombinant SOD fusion protein was expressed and purified with Glutathione sepharose 4B.C57BL/6J mice were immunized with the recombinant SOD fusion protein mixed with Freund adjuvant.Four weeks after the final immunization,the mice of the experiment and control groups were challenged with(45?2)S.japonicum cercariae.All the mice were sacrificed on the forty-fifth day after the challenge to calculate the worm reduction rate and egg reduction rate,and to observe the pathologic changes of liver tissue of the mice.Results The worm reduction rate was 35.63% and the egg reduction rate was 31.17% in the experiment group.The number of granuloma in the live tissue of the experiment group was less than that of the control group,and the mean diameter of single granuloma in the experiment group reduced by 22.32% compared with that of the control group.The IgG subclass levels of IgG1,IgG2a,IgG2b were higher than those of the control group.Conclusion The recombinant SOD fusion protein has a protective effect against Schistosoma japonicum infection.

BACKGROUND:The Y-shaped mesh graft material weaved using lightweight polypropylen has the appropriate porosity, which not only can make the vaginal tissues grow and fuse rapidly on the mesh, but also can maintain good biological strength to ensure the fixed strength for the presacral suspension. OBJECTIVE:To retrospectively analyze the clinical effect of Y-shaped mesh for the biological function reconstruction of the female pelvic floor. METHODS:Ten female cases of pelvic floor dysfunction were enrol ed, aged 37-73 years, and al were given the treatment of sacral colposuspension under laparoscopy. Then perioperative complications were recorded;patients were fol owed up regularly to record the Pelvic Organ Prolapse Quantification (POP-Q) score at different time points;and the subjective satisfaction was investigated using the Pelvic Floor Impact Questionnaire (PFIQ-7). RESULTS AND CONCLUSION:After at least 6-month fol ow-up, no postoperative pelvic infection, nerve damage and complications appeared, the patients healed wel , and no mesh erosion, infection and other adverse reactions occurred. The POP-Q and PFIQ-7 scores at 1, 3 and 6 months after repair were significantly improved than those before repair (P<0.05). These results suggest that the Y-shaped biological mesh repairing female midpelvic floor dysfunction has good biocompatibility, and can restore the anatomy of the pelvic floor.

Background At present, the treatment of silicosis is still limited, and no method is available to cure the disease. miRNAs are involved in the process of fibrosis at the transcriptional level by directly degrading target gene mRNA or inhibiting its translation. However, how miR-130a-3p regulates silicosis fibrosis has not been fully elucidated yet. Objective To investigate whether miR-130a-3p promotes epithelial-mesenchymal transition (EMT) by inhibiting peroxisome proliferators-activated receptors gamma (PPAR-γ), thereby pro-moting the process of silicotic fibrosis. To identify effective new targets for the treatment of silicotic fibrosis. Methods (1) Animal experiments: C57BL/6J mice were intratracheally injected with a one-time dose of 10 mg silica suspension (dissolved in 100 μL saline) as positive lung exposure. A silicosis model group was established 28 d after the exposure. A control group was injected with the same amount of normal saline into the trachea. Hematoxylin-eosin staining and Sirius red staining were used to observe the pathological changes and collagen deposition in lung tissues respectively. Realtime fluorescence-based quantitative polymerase chain reaction (RT-qPCR) was used to assay the expression of miR-130a-3p and PPAR-γ mRNA in lung tissues. Western blotting was used to detect the protein expression of PPAR-γ, transforming growth factor (TGF)-β1, E-cadherin, α-smooth muscle actin (α-SMA), and Collagen Ⅰ in lung tissues. (2) Cells experiments: Mouse lung epithelial cells (MLE-12) were induced with 5 µg·L⁻¹ TGF-β1 for different time (0, 12, 24, 48 h). RT-qPCR was used to detect the expression of miR-130a-3p and PPAR-γ mRNA in cells. The binding relationship between miR-130a-3p and PPAR-γ mRNA was verified by dual luciferase reporter gene assay. MLE-12 cells were stimulated by 5 µg·L⁻¹ TGF-β1 after transfection of miR-130a-3p inhibitor, and Western blotting was used to measure the protein expression of PPAR-γ, E-cadherin, and α-SMA in the TGF-β1-induced cells. Results In the silicosis model group, the alveolar septum was widened and the pulmonary nodules were formed. The Sirius red staining collagen deposition in pulmonary nodules indicated that a silicosis fibrosis model was successfully established. The expressions of TGF-β1, α-SMA, and Collagen Ⅰ proteins were increased, and the expressions of E-cadherin and PPAR-γ proteins were decreased in lung tissues of the silicosis group, compared with the control group (P<0.05 or P<0.01). The expression of miR-130a-3p was increased and the expression of PPAR-γ mRNA was decreased in lung tissues of the silicosis model (P<0.01). The expression of miR-130a-3p was significantly increased, while the expression of PPAR-γ mRNA was decreased in the TGF-β1 induced MLE-12 cells (P<0.05 or P<0.01). The dual luciferase reporter assay showed a direct relationship between miR-130a-3p and PPAR-γ mRNA in MLE-12 cells. The transfection of miR-130a-3p inhibitor in the TGF-β1 induced MLE-12 cells inhibited the decrease of PPAR-γ and E-cadherin proteins, and the increase of α-SMA protein in the MLE-12 cells induced by TGF-β1 (P<0.05 or P<0.01). Conclusion miR-130a-3p promotes the development of silicosis fibrosis by targeting PPAR-γ to increase pulmonary EMT.

Objective To analyze the clinical effect of uterine artery ligation combined with modified uterine compression suture in the prevention and treatment of postpartum hemorrhage.Methods A total of 107 patients with refractory cesarean section who were treated with uterine artery ligation combined with modified uterine compression suture were selected as treatment group.Of the patients treated with conventional treatment,83 patients were included in the control group.The effective hemostasis rate,clinical curative effect,incision infection,postpartum hemorrhage,blood transfusion rate,mattress rate and postoperative morbidity were compared between the two groups.Results The amount of intraoperative blood loss (1 106 ± 458.78) mL in the treatment group was significantly higher than (753.6 ±302.30)mL in the control group,the difference was statistically significant (t =1.892,P ＜ 0.05).The 24h total blood loss of the the treatment group [(1 028.89 ± 375.87) mL] was significantly higher than (813.87 ± 312.15) mL of the control group (t =3.872,P ＜ 0.05).The postoperative 24h blood loss in the treatment group [(70.86 ±19.30)mL] was lower than (73.50 ± 21.20)mL in the control group,and the difference was not statistically significant (t =0.045,P ＞ 0.05).The transfusion rate in the treatment group was 27.10％,which in the control group was higher than 10.84％,the difference was statistically significant between the two groups (x2 =7.722,P ＜ 0.05).The effective hemostasis rate in the treatment group was 99.66％ (106/107),which was significantly higher than 96.39％ in the control group (x2 =6.263,P ＜ 0.05).The rate of puerperal infection was 7.48％ in the treatment group,which in the control group was 6.02％,there was no significant difference between the two groups (P ＞ 0.05).The incision infection rate and postoperative morbidity rate were 0.00％ in the treatment group and the control group,and the two groups were treated with Ⅱ / A healing.Conclusion Uterine artery ligation combined with improved uterine compression suture in the prevention and treatment of postpartum hemorrhage has good hemostasis effect,hysterectomy rate is low,it is worthy of promoting and application.

Objective To analyze the correlation of interferon-induced protein with tetratricopeptide repeats 1(IFIT1)with immune infiltration and prognosis of ovarian cancer(OC).Methods GEO database was employed to select the tumor immune related genes,and Kaplan-Meier and Prognoscan databases were used to identify the genes significantly associated with OC prognosis.The differential expression of IFIT1 between OC tissue and normal tissue were confirmed with GEPIA,Human Protein Atlas,and Timer databases.The expression level of IFIT1 in OC tissues with different grades and stages were analyzed in the UALCAN database.In addition,based on David database,GO enrichment analysis was used to analyze the interacting genes and proteins of IFIT1 in the String and Genemania databases.Timer and Tisidb databases verified the correlation between IFIT1 and immune cells mutually.Finally,after IFIT1 knockdown xenograft model was constructed based on lentiviral vector of IFIT1 shRNA,the tumor growth was observed in the transplanted nude mice,and infiltration of neutrophils was observed with immunohistochemical assay.Results FIT1,a tumor immune gene,selected from the GEO database,Kaplan-Meier and Prognoscan databases,was negatively correlated with the OC prognosis.GEPIA,Human Protein Atlas,Timer database,and UALCAN database indicated that the expression level of IFIT1 was significantly higher in the OC tissues than the normal ovarian tissues,and had no obvious correlation with tumor stage and grade.Analysis in String,Genemania,and David database found the interaction genes and proteins of IFIT1 were enriched in activation of 2'-5'oligonucleotide synthase,virus defense,and innate immunity,and other processes.The Timer database presented that IFIT1 was positively correlated with the infiltration of CD8+T cells,B cells,dendritic cells,neutrophils,and macrophage in OC,with neutrophils having the most significant correlation.Tisidb and GSCA also confirmed the positive correlation between IFIT1 and neutrophil infiltration in OC(P＜0.05).RT-qPCR and Western blotting confirmed that IFIT1 knockdown resulted in suppressed tumor growth in OC cells xenograft nude mice(P＜0.05)and reduced neutrophil infiltration in the xenograft tissues.Conclusion IFIT1 may influence the malignant progression of OC by promoting neutrophil infiltration.

Objective To evaluate prospectively the side effects and tolerance of docetaxel with concurrent late-course hyperfractionated radiotherapy after breast-conserving surgery for stage T1-T2 breast cancer, and to assess the value of this treatment in shortening the treatment time and reducing the economic burden among patients. Methods A total of 20 patients with T1-T2 breast cancer were recruited after they underwent breast-conserving surgery. The acute radiation response classification, treatment completion rate, disease-free survival, hospital stays, and treatment costs were observed. Radiotherapy for all patients was started before the last single-agent docetaxel chemotherapy. Results The completion rate of treatment and the good rate of cosmetic effect reached 100%. The main adverse reactions were hematological toxicity (leukopenia) and skin reactions, which were tolerated. The median follow-up time was 30.1 months, and the follow-up rate was 100%. The average total treatment time of this hyperfractionated radiotherapy with concurrent docetaxel was four weeks, and the total hospitalization cost savings was approximately 10, 000 yuan. The 21-month disease-free survival rate was 100%. Conclusion Stage T1-T2 breast cancer can tolerate hyperfractionated radiotherapy with concurrent chemotherapy after a breast-conserving operation. The procedure results in good local control and satisfactory cosmetic effects, with high health and economic value.

Objective:To investigate the factors affecting the prognosis of stage Ⅰa2-Ⅱa2 cervical cancer after laparoscopic radical hysterectomy (LRH), and to compare the prognosis and recurrence sites of patients with different colpotomy paths.Methods:The clinical data of 965 patients with stage Ⅰa2-Ⅱa2 cervical cancer who underwent LRH in the First Affiliated Hospital of Army Medical University from January 2015 to December 2018 were collected. The median age was 47.0 years of all patients with a median follow-up of 62 months (48-74 months). Cox regression was used to perform the univariate and multivariate analysis of the clinicopathological factors associated with the prognosis that included disease-free survival (DFS) and overall survival (OS). Patients were categorized into LRH through vaginal colpotomy (VC group, n=475) and LRH through intracorporeal colpotomy (IC group, n=490) according to the colpotomic approaches. The prognosis and recurrence sites of patients in each group were compared. Results:(1) During the follow-up period, 137 cases recurred (14.2%, 137/965) and 98 cases died (10.2%, 98/965). The 5-year DFS and OS were 85.8% and 89.9%, respectively. In univariate analysis, positive vaginal margin (PVM) was significantly affected the 5-year OS of patients with cervical cancer ( P=0.023), while clinical stage, maximum diameter of tumor, degree of pathological differentiation, lymph node metastasis (LNM), depth of cervical stromal invasion, parametrium involvement, and uterine corpus invasion (UCI) were significantly associated with 5-year DFS and OS in patients with cervical cancer (all P<0.05). In multivariate analysis, clinical stage ( HR=1.882, 95% CI: 1.305-2.716), LNM ( HR=2.178, 95% CI: 1.483-3.200) and UCI ( HR=3.650, 95% CI: 1.906-6.988) were independent risk factors of 5-year DFS (all P<0.001). Clinical stage ( HR=2.500, 95% CI: 1.580-3.956), LNM ( HR=2.053, 95% CI: 1.309-3.218), UCI ( HR=3.984, 95%C I: 1.917-8.280), PVM ( HR=3.235, 95% CI: 1.021-10.244) were independent risk factors of 5-year OS (all P<0.05). (2) Different colpotomy paths did not significantly affect the 5-year DFS and OS of patients with stage Ⅰa2-Ⅱa2 cervical cancer. The 5-year DFS in VC group and IC group were 85.9% and 85.6% ( P=0.794), and the 5-year OS were 90.8% and 89.3% ( P=0.966), respectively. Recurrence patterns consisted of intraperitoneal recurrence, pelvic recurrence, vaginal stump recurrence, and lymph node and distant metastasis. The intraperitoneal recurrence rate of VC group was significantly lower than that of IC group [0.6%(3/468) vs 2.3% (11/485), P=0.037], while the rates of pelvic recurrence, vaginal stump recurrence, lymph node and distant metastasis and overall recurrence were not significantly different between two groups (all P>0.05). Subgroup analysis of patients with different clinical stages, LNM and UCI showed that statistical differences of the intraperitoneal recurrence rates between two groups were only in patients without LNM (0.5% vs 2.3%, P=0.030) or without UCI (0.7% vs 2.3%, P=0.037). Conclusions:Clinical stage, LNM, PVM and UCI are independent risk factors for the prognosis of patients with stage Ⅰa2-Ⅱa2 cervical cancer. For patients without LNM or UCI, LRH through VC could reduce the intraperitoneal recurrence rate, while it is not enough to improve 5-year DFS and OS of patients. Low proportion of intraperitoneal recurrence, intra-operative tumor cells spillage to vagina stump and pelvic cavity might be the explanation.