OBJECTIVE:To prepare aspirin-β-cyclodextrin-PLGA microspheres,and control its quality. METHODS:Aspirin-β-cy-clodextrin inclusion complexes were firstly prepared,and then aspirin-β-cyclodextrin-PLGA microspheres were prepared by emul-sion-solvent evaporation method. The morphology and particle size of microspheres were detected,and entrapment efficiency and accu-mulative release rate were calculated. With entrapment efficiency as index,orthogonal test was adopted to optimize stirring speed,PVA concentration,PVA volume and feed ratio. RESULTS:The optimal formulation was as follows as stirring speed of 4 000 r/min,PVA concentration of 3%(g/100 ml),PVA volume of 30 ml,feed ratio of 1∶10. Prepared microspheres were round and smooth in appear-ance. Entrapment efficiency of the microspheres was (41.79 ± 1.09)%. The diameter were regular and ranged 0.5-127.5 μm. As drug-loaded microspheres degraded,the release of aspirin was slow and its accumulative release rate was 83%within 600 h. CONCLU-SIONS:Aspirin-β-cyclodextrin-PLGA microspheres are prepared successfully with regular morphology and good sustained-release.

Objective To investigate the correlation between 24 h urinary protein quantitation and pregnancy outcome in patients with pre-eclampsia.Methods A total of 332 pre-eclampsia patients were selected in Tongji Hospital from January 2014 to December 2016.The patients were divided into microalbuminuria group(24 h urinary protein quantification ＜ 0.3 g,n =46),mild proteinuria group (0.3 g ≤ 24 h urinary protein quantification ＜ 2.0 g,n =98),moderate proteinuria group (2.0 g ≤ 24 h urinary protein quantification ＜ 5.0 g,n =71) and severe proteinuria group(24 h urinary protein quantification ≥ 5.0 g,n =117) according to the results of 24 h urinary protein quantification.The pregnancy outcomes were compared between the four groups.Results The 24 h urinary protein quantification and the serum creatinine,urea nitrogen,uric acid levels in the mild proteinuria group,moderate proteinuria group and severe proteinuria group were significantly higher than those in the microalbuminuria group (P ＜ 0.05);and gestational week was significantly shorter than that in the microalbuminuria group (P ＜ 0.05).The 24 h urinary protein quantification and serum urea nitrogen,uric acid levels in the moderate proteinuria group were significantly higher than those in the mild proteinuria group (P ＜ 0.05);and gestational week was significantly shorter than that in the mild proteinuria group (P ＜ 0.05);but there was no significant difference in serum creatinine level between the two groups (P ＞ 0.05).The 24 h urinary protein quantification,serum creatinine,urea nitrogen and uric acid levels in the severe proteinuria group were significantly higher than those in the mild proteinuria group (P ＜ 0.05);and the gestational week was significantly lower than that in the mild albuminuria group (P ＜ 0.05).The 24 h urinary protein quantification in the severe proteinuria group was significantly higher than that in the moderate proteinuria group (P ＜ 0.05),but there was no significant difference in the gestational week and serum creatinine,urea nitrogen,uric acid levels between the two groups (P ＞0.05).There was no significant difference in the rates of cesarean section and spontaneous labor between the four groups (P ＞0.05).The rate of induced labor in the moderate proteinuria group and the severe proteinuria group was significantly higher than that in the mild albuminuria group and the microalbuminuria group (P ＜ 0.05).There was no significant difference in the rate of induced labor between the mild proteinuria group and the microalbuminuria group (P ＞ 0.05).There was no significant difference in the rate of induced labor between the severe proteinuria group and the moderate proteinuria group (P ＞ 0.05).The incidence of complications in microalbuminuria group,mild proteinuria group,moderate proteinuria group and severe proteinuria group was 30.43％ (14/46),47.96％ (47/98),74.65％ (53/71) and 74.36％ (87/117) respectively;the incidence of complications in the moderate proteinuria group and the severe proteinuria group was significantly higher than that in the microalbuminuria group and the mild albuminuria group (P ＜ 0.05),but there was no significant difference in the incidence of complications between microalbuminuria group and mild albuminuria group (P ＞ 0.05),there was no significant difference in the incidence of complications between the moderate proteinuria group and the severe proteinuria group (P ＞0.05).The incidences of premature birth and neonatal asphyxia in the mild proteinuria group were significantly higher than that in the microalbuminuria group (P ＜ 0.05),and the body mass of the neonates was significantly lower than that in the microalbuminuria group (P ＜0.05),but there was no significant difference in the perinatal mortality rate and the incidences of fetal growth restriction(FGR) and poor neonatal resuscitation between the two groups (P ＞ 0.05).The incidences of FGR,premature birth,neonatal asphyxia,poor neonatal resuscitation and the perinatal mortality in the moderate proteinuria group and severe proteinuria group were significantly higher than those in the microalbuminuria group (P ＜ 0.05);and neonatal body mass was significantly lower than that in the mieroalbuminuria group (P ＜ 0.05).The incidences of FGR,premature birth and poor neonatal resuscitation and perinatal mortality in the moderate proteinuria group were significantly higher than those in the mild proteinuria group (P ＜ 0.05);and the neonatal body mass was significantly lower than that in the mild proteinuria group (P ＜ 0.05);but there was no significant difference in the neonatal asphyxia incidence between the two groups (P ＞ 0.05).The incidences of FGR,premature birth,neonatal asphyxia,poor neonatal resuscitation and perinatal mortality in the severe proteinuria group were significantly higher than those in the mild proteinuria group (P ＜ 0.05);and the body mass of the newborns was significantly lower than that in the mild albuminuria group (P ＜ 0.05).The incidence of neonatal asphyxia in the severe proteinuria group was significantly higher than that in the moderate proteinuria group (P ＜ 0.05),but there was no significant difference in the incidences of FGR,premature birth,poor neonatal resuscitation,perinatal mortality and neonatal body mass between the two groups (P ＞ 0.05).Conclusions The of 24 h urinary protein quantitation is closely related to the pregnancy outcome in patients with pre-eclampsia,the 24 h urinary protein quantification should be regularly detected in the patients with pre-eclampsia.When the urinary protein quantitation is more than 2.0 g,the incidences of maternal complications and poor prognosis of the perinatal infants is significantly higher,but the boundary value of the 24 h urinary protein quantitation for the diagnosis of severe pre-eclampsia still needs further large sample study.

The prevalence of human papilloma virus (HPV)-16 in patients with cervical cancer, the physical status of HPV-16 in patients with cervical lesions, and the role of HPV-16 integration in cervical carcinogenesis were investigated. HPV genotyping was performed by using PCR approach with the primer GP5+/GP6+ and type-specific primer on biopsy specimens taken operatively from 198 women. Multiple PCR was done to detect physical status of HPV-16 in a series of cervical liquid-based cytology samples and biopsy specimens obtained from different cervical lesions with HPV-16 infection, including 112 specimens with cervical cancer, 151 specimens with CIN I, 246 specimens with CIN and 120 specimens with CINIII. The results showed that there were 112 cervical cancer samples (56.57% of total cervical cancer patients) with HPV-16 infection. The frequency of HPV-16 pure integration was 65.18% (73/112), 56.57% (47/120), 23.58% (58/246) and 7.95% (12/151) in cervical cancer, CINIII, CINII and CINI patients respectively. In situ hybridization was performed on some paraffin-embedded sections of CINII, CINIII and cervical cancer to verify the physical status of HPV-16 infection. Significant difference was observed between cervical cancer and CIN I, CINII, CINIII in the frequency of HPV-16 integration (P<0.01). It is suggested that HPV-16 is the most prevalent type and is associated with cervical cancer. In the case of HPV-16 infection there are close associations between the severity of cervical lesions and the frequency of HPV-16 integration. The application of testing HPV genotyping and physical status based on detection of HC-II HPV DNA would be in favor of predicting the prognosis of cervical precancerosis and enhancing the screening accuracy of cervical cancer.

Objective To investigate the renal pathologic changes in both donors and transplant recipients with delayed graft function (DGF).Methods The clinical and laboratory data were retrospectively analyzed in 144 renal recipients with DGF.All the patients received renal biopsy,and donors' biopsy was performed on 131 recipients.The pathological changes were examined under the light microscopy (LM),immunofluorescence (IF) and electron microscopy (EM).Results (1) The incidence of DGF was 10.16%-7.48% during 1994 to 2005,and decreased to 5.35 % during 2006 to 2009.(2) Anuria occurred in 24 cases (16.67 %),oliguria in 24 (16.67%) and hypertension in 68 cases (47.22 %).The enlargement of transplanting kidney and the increased vascular resistance was detected in 79.67 % (98/123 cases) and 45.53 % (56/123 cases) respectively by ultrasound examination.(3) The level of serum creatinine was ranged from 451 to 707 μmol/L.The high level of urinary NAG enzyme was found in 102 cases (70.83 %),proteinuria in 79 recipients (54.86 %) and hematuria in 77 cases (53.47 %).(4) The acute rejection was observed in 66 cases (45.83 %),toxicity of CNI in 22 (15.28 %),IgA nephropathy in 18 (12.50 %),acute tubular necrosis in 8 (5.56 %),and recurrent FSGS in 2 cases (1.39 %).(5) In most recipients (66/109 cases,60.55 %)immunosuppressive regimen altered and renal replacement therapy was given.Conclusion The causes of DGF are complicated.The quality of donors' kidney and early histological changes of recipients are contributed to the development of DGF.It is necessary to perform renal biopsy not only in donors but also in recipients with DGF.And kidney biopsy in transplanted patients was also beneficial to the treatment.

The aim of the present study was to examine the effects of suppression of EphB4 and/or mTOR on the biological behaviors of ovarian cancer cells, and the potential regulatory pathways. Antisense EphB4 vectors and shRNA vectors targeting mammalian target of rapamycin (mTOR) were constructed and transfected into A2780 and SKOV3 cells (two ovarian cancer cell lines). The effects of the antisense EphB4 vectors and the shRNA vectors on the proliferation, apoptosis and invasion of ovarian cancer cells were measured, and the expression of EphB4, mTOR and Akt detected. The results showed that transfection with mTOR shRNA could inhibit growth, induce apoptosis, and reduce invasive ability of ovarian cancer cells, which was accompanied by downregulation of EphB4, mTOR and Akt. The inhibitory effects on cell growth caused by mTOR shRNA alone were weaker than those by antisense pEGFP-C1-EphB4. In the antisense pEGFP-C1-EphB4-transfected cells, it was found that EphB4 knockdown could decrease the mTOR expression and slightly reduce the Akt phosphorylation. Significant suppressive effects on cell growth were observed in cells co-transfected with antisense pEGFP-C1-EphB4 and mTOR shRNA. In co-transfection group, the expression levels of EphB4, mTOR and Akt were distinctly lower than those in other groups. It was concluded that suppression of EphB4 may inhibit the growth of ovarian cancer cells by downregulation of the PI3K/Akt/mTOR pathway, and reverse Akt phosphorylation induced by mTOR shRNA. Inhibition of EphB4 and mTOR combined may cooperatively suppress the biological behaviors of ovarian cancer cells.

Objective To introduce the detailed surgical procedure of parachute-like-suture double-pouch pancreaticojejunostomy in laparoscopic pancreaticoduodenectomy (LPD),and to study its clinical results.Methods A retrospective study was conducted on 31 patients who underwent parachute-like-suture double-pouch pancreaticojejunostomy in laparoscopic pancreaticoduodenectomy between January 2016 and June 2018 in our department.Results Of 31 patients who underwent total laparoscopic pancreaticoduodenectomy,5 patients underwent two-dimensional (2D) LPD and 26 underwent three-dimensional (3D) LPD.Total mesopancreas excision (TMpE) was performed in 12 patients,including 2 patients combined with PV-SMV segmental resection and reconstruction.The mean operative duration,and mean estimated blood loss,post-operative hospital stay were 412.8 ± 102.4 min,462.8 ± 396.7 ml,14.7 ± 8.9 d,respectively.The operating time of parachute-like-suture double-pouch pancreaticojejunostomy was 29.7 ± 6.8 min (20 ～45 min).Post-operative complications occurred in 8 patients (25.8％),3 of whom suffered from more than two types of complications.There were 2 patients (6.5％) with postoperative B/C grade pancreatic fistula,4 patients with postoperative biliary leakage (12.9％),3 patients with delayed gastric emptying (9.7％),1 patient with portal vein thrombosis (3.2％) and 1 patient with peritoneal effusion (3.2％).One patient died during perioperative period due to gastrointestinal hemorrhage.Conclusions Parachute-like-suture double-pouch pancreaticojejunostomy in laparoscopic pancreaticoduodenectomy was a simple,convenient,reliable method.It is worthy of clinical promotion and further studied.

Objective:To summarize the technical points of laparoscopic pancreaticoduodenectomy (LPD) carried out in a single center.Methods:The clinical data of 102 patients who underwent laparoscopic pancreaticoduodenectomy in 2018 at the Department of Hepatobiliary Surgery of the Second Hospital of Hebei Medical University were retrospectively analyzed, and the general conditions, operation time, intraoperative blood loss, and surgical operation were analyzed. Post-operative complications, hospital stay and other indicators were studied.Results:Of the 102 patients who were included, there were 57 males and 45 females, aged 15.0 to 79.0 (59.9±11.8) years old, with a body mass index (23.6±3.6) kg/m 2. For the 102 patients who underwent LPD, 6 were total pancreatic resection. Three were combined with vascular resection in the form of portal vein-superior mesentery vein segmental resection. The operation time was (376.6±87.2) min, the intraoperative blood loss was 350 (100, 800) ml, and the postoperative hospital stay was (17.0±5.9) days. Postoperative complications occurred in 26 of the 102 patients (25.5%), and more than two complications occurred in 17 patients. B/C grade pancreatic fistula occurred in 9 patients (9.4%), abdominal bleeding in 8 patients (7.8%), gastrointestinal bleeding in 2 patients (2.0%), biliary fistula in 4 patients (3.9%), and gastric emptying disorder in 5 patients (4.9%), 8 patients had pulmonary infection (7.8%). Five patients (4.9%) died during the perioperative period. Conclusion:The main technical points of LPD included en bloc resection, pancreaticojejunostomy, and vascular reconstruction. The basis of LPD is en bloc resection. Combined resection and reconstruction of vascular segments is a sign of maturity of LPD technology and a prerequisite for further development as a routine procedure.

The aim of the present study was to examine the effects of suppression of EphB4 and/or mTOR on the biological behaviors of ovarian cancer cells, and the potential regulatory pathways. Antisense EphB4 vectors and shRNA vectors targeting mammalian target of rapamycin (mTOR) were constructed and transfected into A2780 and SKOV3 cells (two ovarian cancer cell lines). The effects of the antisense EphB4 vectors and the shRNA vectors on the proliferation, apoptosis and invasion of ovarian cancer cells were measured, and the expression of EphB4, mTOR and Akt detected. The results showed that transfection with mTOR shRNA could inhibit growth, induce apoptosis, and reduce invasive ability of ovarian cancer cells, which was accompanied by downregulation of EphB4, mTOR and Akt. The inhibitory effects on cell growth caused by mTOR shRNA alone were weaker than those by antisense pEGFP-C1-EphB4. In the antisense pEGFP-C1-EphB4-transfected cells, it was found that EphB4 knockdown could decrease the mTOR expression and slightly reduce the Akt phosphorylation. Significant suppressive effects on cell growth were observed in cells co-transfected with antisense pEGFP-C1-EphB4 and mTOR shRNA. In co-transfection group, the expression levels of EphB4, mTOR and Akt were distinctly lower than those in other groups. It was concluded that suppression of EphB4 may inhibit the growth of ovarian cancer cells by downregulation of the PI3K/Akt/mTOR pathway, and reverse Akt phosphorylation induced by mTOR shRNA. Inhibition of EphB4 and mTOR combined may cooperatively suppress the biological behaviors of ovarian cancer cells.
Apoptosis ; genetics ; Cell Line, Tumor ; Cell Proliferation ; Down-Regulation ; genetics ; Female ; Humans ; Ovarian Neoplasms ; pathology ; physiopathology ; Proto-Oncogene Proteins c-akt ; genetics ; metabolism ; RNA, Small Interfering ; genetics ; Receptor, EphB4 ; genetics ; metabolism ; Suppression, Genetic ; genetics

Objective To review our experience in laparoscopic pancreaticoduodenectomy combined with major vascular resection and reconstruction.Methods Of 183 patients who underwent laparoscopic pancreaticoduodenectomy in our department from November 2013 to January 2017,major vascular resection and reconstruction using the SMA first approach for total mesopancreas excision was performed in 7 patients.The clinical data of these 7 patients were retrospectively analyzed.Results Total 3D laparoscopic surgery was performed in all these 7 patients.The mean operation time,mean blood loss and blood flow occlusion time were (551.4 ± 83.8) min,(671.3 ± 256.3) ml and (45.8 ± 6.7) min,respectively.Six out of 7 patients were pathologically diagnosed to have pancreatic adenocarcinoma with negative surgical margins.Two patients had lymphatic metastasis (the number of metastatic lymph node was 1 in each patient).The mean number of lymph nodes resected was (12.7 ± 5.8).The portal vein-superior mesenteric vein (PV-SMV) was segmentally resected and reconstructed using an end to end anastomosis following the preoperative plan in 4 patients.These included 2 patients who underwent total pancreatectomy.The portal vein was wedge-resected and reconstructed by venorrhaphy in 2 patients.The remaining 1 patient was histopathologically diagnosed to have a mass-type chronic pancreatitis.Only 1 patient was treated in the ICU for 1 day after surgery.Post-operative complications occurred in 2 patients and they were managed with nonsurgical treatment (PV-SMV thrombosis and gastric emptying disorder in 1 and a pancreatic leakage (level A) in 1).The mean length of post-operative hospital stay was (13.7 ± 3.2) days with no in-hospital mortality.Seven patients were alive by April 2017.The mean follow-up for the 6 patients with pancreatic adenocarcinoma was 4.5 (3.5 ～9) months.Conclusions Based on our experience in skillful and masterly major vascular resection and reconstruction in open surgery and on our experience in standard laparoscopic pancreaticoduodenectomy,laparoscopic pancreatieoduodenectomy combined with major vascular resection and reconstruction was feasible and safe.This surgery requires very mature skills in laparoscopic surgery.

Objective: To compare the difference of the clinical and laboratory characteristics between γδ T-cell large granular lymphocyte leukemia (γδT-LGLL) and αβ T-cell large granular lymphocyte leukemia (αβT-LGLL) . Methods: The clinical and laboratory characteristics of 17 patients with γδT-LGLL and 91 patients with αβT-LGLL in the department of therapeutic center of anemia of enrolled in our hospital from January 2009 to January 2019 were retrospectively analyzed. Results: The median age of the 17 patients with γδT-LGLL was 54 years (range, 25-73 years) , the most common presenting symptom was anemia. In comparison with αβT-LGLL patients, splenomegaly was common (41% and 44%, respectively) , whereas hepatomegaly (12% and 5%, respectively) and lymphadenopathy (6% and 8%, respectively) were rare. The positive rates of antinuclear antibody (59% and 45%, respectively) were high, whereas the positive rates of rheumatoid factor (6% and 10%, respectively) were rare for both groups. There were no differences on peripheral blood counts between the two groups. However, γδT-LGLL patients were found to be predominantly expressed a CD4⁻/CD8⁻ phenotype. Steroid therapy with prednisone was used alone as first-line therapy for 1 patient. Cyclosporin A (CsA) was used alone as first-line therapy for 3 patients. CsA in combination with steroids were administered in 13 patients. After 4 months treatment, 2 patients acquired complete response, 4 patients acquired partial response, the overall response was 35%. Conclusion γδT-LGLL is a rare mature T-lymphocyte proliferative disease. Clinical and laboratory characteristics were quite similar for γδT-LGLL in compare with αβT-LGLL. γδT-LGLL predominantly expressed a CD4⁻/CD8⁻ phenotype. The data presented here indicate the CsA is an effective option for the first-line treatment of γδT-LGLL.