Parathyroid hormone related protein (PTHrP) has important biological functions in calcium metabolism. The aim of this study was to silence the expression of PTHrP by RNA interference and recombinant adenovirus, and to provide a material to investigate the relative functions of PTHrP in goat mammary gland epithelial cell. The Block-iT shRNA interference system was used in this experiment. We designed and synthesized two pairs of complementary single-strand DNA oligonucleotides (shRNA-322/357) targeting two different sites of PTHrP mRNA. Then the oligonucleotides were inserted into shuttle vector pENTR/CMV-GFP/U6. After detection of the interference efficiency by Western blotting, we chose pENTR/CMV-GFP/U6-322 and adenovirus backbone vector pAD/PL-DEST to produce recombinant vector pAD/PL-DEST/CMV-GFP/U6-322. The first generation recombinant adenovirus particles (AD-PTHrP-322) were produced and further amplified by transfecting HEK-293 cells. The titer of the recombinant adenovirus reached 2.0 x 1(9) PFU/mL determined by TCID50 assays. The result of real-time quantitative PCR indicated that mRNA expression levels of gene were reduced 29.2%, 68.1% and 82.6% (P < 0.05), respectively, when goat mammary gland epithelial cells were infected with AD-PTHrP-322 after 24, 48 and 72 h, in which PTHrP. Western blotting also showed that the expression of PTHrP was reduced by infecting the cells with AD-PTHrP-322. AD-PTHrP-322 has been proved with significant interference effect on expression of PTHrP.
Adenoviridae ; genetics ; metabolism ; Animals ; Epithelial Cells ; metabolism ; Female ; Genetic Vectors ; genetics ; Goats ; HEK293 Cells ; Humans ; Mammary Glands, Animal ; cytology ; Parathyroid Hormone-Related Protein ; biosynthesis ; genetics ; RNA Interference ; RNA, Messenger ; biosynthesis ; genetics ; RNA, Small Interfering ; genetics ; Recombinant Proteins ; biosynthesis ; genetics ; isolation & purification ; Transfection

deltaFosB, a naturally occurring truncated isform of fosB gene, existed in many tissues stably and played an important role in formation and differentiation of adipocyte and osteoblast. deltaFosB may be related to the metabolism of calcium in bone and mammary gland and regulate the signal pathway of calcium transfer from bone to mammary gland. We first sub-cloned deltafosB gene of goat into the vector pET32a to construct prokaryotic expression vector pET32a-deltafosB. Then we induced for deltafosB gene expression efficiently by IPTG. Finally we immunized the adult rabbits with purified recombinant deltaFosB to prepare rabbit anti-goat deltaFosB polyclonal antibody. iELISA analysis showed the antibody with the titer of 1:51 200, and Western blotting result showed that the antibody could specifically detect the deltaFosB protein expressed in prokaryotic cell and HEK-293 cell, respectively. Further Western blotting assay showed that deltaFosB expressed in various tissues of goat in vivo.
Animals ; Antibodies, Monoclonal ; biosynthesis ; Cloning, Molecular ; Escherichia coli ; genetics ; metabolism ; Gene Expression ; Goats ; Proto-Oncogene Proteins c-fos ; biosynthesis ; genetics ; immunology ; Rabbits ; Recombinant Proteins ; biosynthesis ; genetics ; immunology

Objective:To initially investigate whether simultaneous radiochemotherapy with hyperthermia can prolong the survival of glioblastoma (GBM) patients.Methods:Clinical data of 61 GBM patients undergoing surgery in our hospital from September 2016 to June 2019 were retrospectively analyzed. According to different treatment methods, all patients were divided into the control group ( n=34) and observation group ( n=27). In the control group, three-dimensional radiotherapy with a dose of 60 Gy combined with temoazolamine chemotherapy was delivered. In the observation group, simultaneous radiochemotherapy with 15-20 cycles of hyperthermia at 40-41℃ was supplemented. The survival time was calculated by Kaplan-Meier method, and the survival time was compared with log-rank test between two groups. Results:The median progression-free survival in the observation group was significantly longer than that in the control group (14.33 months vs.9.94 months, P<0.05). The median overall survival in the observation group was also remarkably higher than that in the control group (18 months vs. 14 months, P<0.05). Conclusions:Simultaneous radiochemotherapy with hyperthermia is innovatively applied to treat GBM after surgical resection. Preliminary findings demonstrate that compared with chemoradiotherapy, simultaneous radiochemotherapy with hyperthermia can prolong the survival time of GBM patients.

Objective:To investigate the factors related to the prognosis of patients with metastatic renal cell carcinoma after bone metastasectomy and to provide a reference for the clinical treatment of renal cell carcinoma.Methods:The clinical data of 143 patients with metastatic renal cell carcinoma in our center from January 2008 to December 2018 were retrospectively collected. Among 143 patients, 121 were male and 22 were female, with the average age of (54.8±12.5) year-old(from 18 to 85 year-old). The KPS scores of 138 patients were no higher than 80%. According to International Metastatic RCC Database Consortium (IMDC) risk model for metastatic renal cell carcinoma, the patients were divided into intermediate risk group (92 patients) and poor risk group (51 patients). The patients who had received complete resection for both primary lesion and metastatic tumor were regarded as tumor-free (47 patients). Otherwise, the patients with unresectable primary tumor or incomplete resection of the metastatic tumor were defined as the patients surviving conversely bone metastases were the only metastatic site, and the other 57 patients also experienced concomitant metastases comparatively. Sixty-two patients only had solitary bone metastasis lesion and 81 patients had multiple bone metastases. Kaplan-Meier survival analysis was used to calculate the 1-year, 3-year and 5-year survival rate. The impacts of different variables on the prognosis were examined by log-rank test. Univariate analysis and multivariable Cox proportional hazards regression models were used to identify the independent risk factors.Results:The median follow-up time was 49.0 months. The overall survival was 1-115 months, with the median OS was 24.0 months. The 1-year, 3-year and 5-year survival rate were 79.2%, 59.4% and 31.6%, respectively. All the patients underwent bone metastasectomy and 72 of them were treated with targeted therapy. The pathology results of metastasectomy were clear cell carcinoma for 132 patients and non-clear cell carcinoma for the 11 patients. In tumor-free group, the median OS was 30.0 months and in survival with tumor group, the median OS was 19.4 months, with a significant difference between the two groups ( P=0.030). In IMDC intermediate risk group, the prognosis was improved among the patients who received post-surgical targeted therapy after metastasectomy (24.3 months vs. 16.8 months, P=0.027), whereas the difference was not significant for IMDC poor risk group ( P=0.449). Age ≥60 years and multiple bone metastases sites were proved to be the independent risk factors for the prognosis of patients with metastatic renal cell carcinoma after bone metastasectomy. Conclusions:The prognosis of RCC patients with bone metastases was generally poor. Metastasectomy could prolong the OS of the patients who had undergone primary nephrectomy and had solitary bone metastasis. Metastasectomy combined with targeted therapy could significantly improve the prognosis of the IMDC intermediate risk patients. However, the effect of targeted therapy among IMDC poor risk patients remained to be further proved.

Objective To compare the perioperative parameters and renal function in patients,whose aged was 65 year-old or above,with clinical T1-2 renal tumors undergoing partial nephrectomy (PN) or radical nephrectomy (RN).Methods A retrospective review of 469 patients,who underwent RN and PN in our center,was conducted from January 2012 to November 2018,icluding 247 in the RN group and 222 in the PN group.The RN group consisted of 170 male and 77 female patients,with the mean age of (70.96 ± 5.21) year-old.126 cases were found that the tumor located on the fight side,with the median diameter of (4.93 ± 2.03) cm.The median BMI,median R.E.N.A.L.score and pre-operation eGFR of the RN group were (24.4 ± 3.1) kg/m2,8.39 ± 1.45) and (80.23 ± 15.14) ml/(min · 1.73 m2),respectively.The PN group consisted of 150 male and 72 female patients,with the mean age of (70.23 ± 4.62) years old.108 patients had tumors on the left side while 114 on the right side,with the median diameter of (3.17 ± 1.41) cm.The median BMI,median R.E.N.A.L score and pre-operation eGFR of the PN group were (23.5 ± 3.2) kg/m2,(6.69 ± 1.81) and (82.83 ± 14.36) ml/(min · 1.73m2),respectively.No statistical difference was noticed in the age,gender,tumor location and BMI between RN group and PN group(P ＞ 0.05).The PN group had smaller tumors and lower R.E.N.A.L.scores than the RN group (P ＜ 0.05).Various parameters were compared between the PN and RN groups,including operative duration,surgical procedure,intraoperative blood loss,perioperative blood transfusion,drainage tube removal time,postoperative duration of hospitalization,pathological results,the renal function immediately after surgery and at 1 month,1 year,2 years,3 years,4 years,5 years after surgery,and the incidence of chronic renal dysfunction.Results Significant differences were found in multiple variables between the two cohorts,such as operative duration [(115.70 ± 39.69) min in RN vs.(132.26 ± 49.02) min in PN],estimated intraoperative blood loss [(45.85 ± 55.93) days in RN vs.(66.60 ± 61.55) ml in PN],drainage tube removal time [(4.38 ± 1.71) days in RN vs.(4.86 ± 1.61) days in PN],duration of postoperative hospitalization [(5.14 ± 1.65) days in RN vs.(5.52 ± 1.32) days in PN] (P ＜ 0.05).Furthermore,higher proportion of RCC was detected in the RN cohort (93.5％ in RN vs.86.5 ％ in PN,P ＜ 0.05).There was no significant difference in perioperative blood transfusion rate between the two cohorts (10.93％ vs.9.01％,P ＞ 0.05).Compared with the RN cohort,the PN cohort had higher eGFR immediately after surgery [(74.08 ± 18.31) ml/(min · 1.73m2) vs.(52.58 ± 14.21) ml/(min · 1.73m2)],1 month after surgery [(76.11 ± 18.34) ml/(min · 1.73m2) vs.(53.78 ± 15.03)ml/(min · 1.73m2)] and at the last follow-up [(73.92 ± 18.59) ml/(min · 1.73m2) vs.(52.35 ± 16.13) ml/(min · 1.73m2)] (P ＜ 0.001).Compared with those of the RN cohort,the incidences of eGFR ＜ 45 ml/(min · 1.73m2) of the PN cohort were lower immediately after surgery [9.01％ (20/222) vs.31.9％ (79/247)],1 month after surgery [7.87％ (14/178) vs.27.31％ (62/227)],1 year after surgery [8.96％ (13/145) vs.38.75％ (62/16 0)],2 years after surgery [9.89％ (9/91) vs.31.57％ (42/133)],3 years after surgery [13.21％ (7/53) vs.30.61％ (30/98)],4 years after surgery [16.21％ (6/37) vs.30.26％ (23/76)] and 5 years after surgery [18.18％ (4/22) vs.31.11％ (14/45)] (P ＜ 0.001).Conclusion The perioperative risk of PN in the treatment of elderly patients aged 65 and above with clinical cT1-2 renal tumor is controllable.PN could better retain renal function for those patients and reduce the risk of postoperative chronic renal insufficiency.

Objective:To investigate the feasibility and safety of a novel single-port robotic surgical system with flexible 3D endoscope and deformable surgical instruments in zero ischemic partial nephrectomy.Methods:From May 2021 to October 2021, a prospective study on patients with renal tumor who plan to receive zero ischemic partial nephrectomy was conducted. Inclusion criteria included over 18 years old, body index between 18.5 to 30.0 kg/m2, American Society of Anesthesiologists Score ranged from 1 to 3 points, cooperation with the follow-up and related examinations, voluntary in participating the clinical trial and signing the informed consent. Exclusion criteria included patients with other co-existed malignant tumors or a medical history of other malignant tumors, the patients who have received the same urological surgery in the past, the patients who underwent or plan to undergo other major operations 3 months before or after the surgery, the patients with active pulmonary tuberculosis or severe systemic diseases, the patients to be considered not suitable to enroll in by the researchers. A novel single-port robotic surgical system was used to perform the surgery. The system consiststed of a remote control console, a surgical equipment cart, a four-arm operation cart and deformable robotic instruments which were reusable. The two-section deformable robotic instruments were able to bend in four directions and carried different surgical instruments such as unipolar scissors, bipolar grasping forceps and needle holders. The deformable robotic instruments entered the body through a special trocar with single hole and multi-channel, and then unfolded in a triangle. By bending instruments, surgeons could perform single-port robotic surgery without the trouble of "chopstick effect" or "reverse direction" . Four cases of single-port partial nephrectomy were carried out. Under general anesthesia, the patients were taken the lateral recumbent position, with elevated waist and lowered head and feet. A 3-4 cm incision was taken layer by layer along the lateral edge of the rectus abdominis at the umbilicus level. A special 2.5 cm robotic trocar was set into the cavity, and a high-definition 3D laparoscopic lens and a snake shaped mechanical arm were then put into the abdominal cavity through the trocar. All operations were performed by transperitoneal approach with an auxiliary port through the same or a different skin incision if necessary. Tumor resection and renal reconstruction were performed by the way of zero ischemia. The perioperative parameters such as tumor size, operation time, intraoperative bleeding and postoperative complications were analyzed.Results:Four patients were involved, including 2 males and 2 females, with 2 of them having a history of hypertension. The ECOG scores were all 0, and KPS score was 100 in 3 cases and 90 in 1 case. Preoperative mean serum creatinine was (76.8±18.8)μmol/L (range 70-104 μmol/L). The tumors were located on the left in 3 cases and on the right in 1 case. The diameter of the tumor ranged from 1.1 to 2.8 cm, with the TNM classification of T 1a. The R. E.N.A.L. scores were 4a, 7p, 6p and 4P respectively. The first operation was performed by pure single-port surgery, and the other 3 cases were performed with the help of an auxiliary port to ensure the safety .The operation time ranged from 155 min to 210 min, and the intraoperative bleeding ranged from 20 ml to 170 ml. Postoperative pathology showed 2 cases of renal clear cell carcinoma with negative margin and 2 cases of angiomyolipoma. No severe complications, such as bleeding or urinary leakage, were observed during the perioperative period, and the change of serum creatinine was insignificant before discharge and before operation( P=0.24). Conclusions:A robotic single-port partial nephrectomy can be successfully carried out by using this novel single-port robotic surgical system with flexible 3D endoscope and deformable surgical instruments.

Objective To explore the scan technique and image quality of coronary artery imaging with dual-source CT without oral Betaloc preparation.Methods Plain and enhanced dual-source CT coronary artery imaging without oral Betaloc preparation was performed in 215 patients with clinically suspected coronary heart disease or early-stage coronary lesions.Calcium scoring with plain scan images and multi-planar reconstruction(MPR),maximum intensity projection(MIP)and volume rendering technique (VRT)reconstruction with enhanced scan images were made in all cases.The scan technique and post reconstruction experience was summarized.The image quality was classified as three grades,and coronary segments classified according to AHA standards were evaluated.Results The median of calcium score of the 215 cases was 82.2(2.3—1827.9).The average heart rate of the enhanced scan was(80.6?15.3) (57—139)bpm.The post reconstruction methods with which coronary segments could be shown as best as possible consisted of(1)multiphases screening methods,(2)bi-phase or multiple-phase complement method,and(3)premature beat removing or arrhythmia shifting method.Altogether 3026 coronary segments were evaluated,among them 97.5% were evaluated as grade 1 image quality,2.0% were evaluated as grade 2 and 0.5% were evaluated as grade 3.The coronary segments in 91 cases were completely normal, while 112 segments with

Objective:To evaluate the clinical characteristics and prognosis of gallbladder cancer (GBC) patients in China.Methods:This retrospective multicenter cohort study enrolled 3 528 consecutive GBC patients diagnosed between January 2010 to December 2017 in 15 hospitals from 10 provinces. There were 1 345 (38.12%) males and 2 183 (61.88%) females.The age of diagnosis was (63.7±10.8) years old (range: 26 to 99 years old) .There were 213 patients (6.04%) in stage 0 to Ⅰ, whereas 1 059 (30.02%) in stage Ⅱ to Ⅲ, 1 874 (53.12%) in stage Ⅳ, and 382 (10.83%) unavailable. Surgery was performed on 2 255 patients (63.92%) . Three hundred and thirty-six patients received chemotherapy or radiotherapy (9.52%; of which 172 were palliative); 1 101 (31.21%) received only supportive treatment.The patient source, treatment and surgery, pathology, concomitant gallstone, and prognosis were analyzed.Results:Among the 3 528 GBC patients, 959 (27.18%) were from East China, 603 (17.09%) from East-North China, 1 533 (43.45%) from Central China, and 433(12.27%) from West China. Among the 1 578 resectable tumor, 665 (42.14%) underwent radical surgery, 913 (57.86%) underwent surgery that failed to follow the guidelines.Eight hundred and ninety-one (56.46%) patients were diagnosed before surgery, 254 (16.10%) during surgery, and 381 (24.14%) after surgery (time point of diagnosis couldn′t be determined in 52 patients) .Among the 1 578 patients with resectable tumor, 759 (48.10%) had concomitant gallstone.Among the 665 patients underwent radical surgery, 69 (10.4%) showed positive resection margin, 510 (76.7%) showed negative resection margin, and 86 (12.9%) unreported margin status.The 5-year overall survival rate (5yOS) for the 3 528-patient cohort was 23.0%.The 5yOS for patients with resectable tumor was 39.6%, for patients with stage ⅣB tumor without surgery was 5.4%, and for patients with stage ⅣB tumor underwent palliative surgery was 4.7%.Conclusions:More than half GBC patients in China are diagnosed in stage Ⅳ.Curative intent surgery is valuable in improving prognosis of resectable GBC.The treatment of GBC needs further standardization.Effective comprehensive treatment for GBC is in urgent need.

Objective:To evaluate the clinical characteristics and prognosis of gallbladder cancer (GBC) patients in China.Methods:This retrospective multicenter cohort study enrolled 3 528 consecutive GBC patients diagnosed between January 2010 to December 2017 in 15 hospitals from 10 provinces. There were 1 345 (38.12%) males and 2 183 (61.88%) females.The age of diagnosis was (63.7±10.8) years old (range: 26 to 99 years old) .There were 213 patients (6.04%) in stage 0 to Ⅰ, whereas 1 059 (30.02%) in stage Ⅱ to Ⅲ, 1 874 (53.12%) in stage Ⅳ, and 382 (10.83%) unavailable. Surgery was performed on 2 255 patients (63.92%) . Three hundred and thirty-six patients received chemotherapy or radiotherapy (9.52%; of which 172 were palliative); 1 101 (31.21%) received only supportive treatment.The patient source, treatment and surgery, pathology, concomitant gallstone, and prognosis were analyzed.Results:Among the 3 528 GBC patients, 959 (27.18%) were from East China, 603 (17.09%) from East-North China, 1 533 (43.45%) from Central China, and 433(12.27%) from West China. Among the 1 578 resectable tumor, 665 (42.14%) underwent radical surgery, 913 (57.86%) underwent surgery that failed to follow the guidelines.Eight hundred and ninety-one (56.46%) patients were diagnosed before surgery, 254 (16.10%) during surgery, and 381 (24.14%) after surgery (time point of diagnosis couldn′t be determined in 52 patients) .Among the 1 578 patients with resectable tumor, 759 (48.10%) had concomitant gallstone.Among the 665 patients underwent radical surgery, 69 (10.4%) showed positive resection margin, 510 (76.7%) showed negative resection margin, and 86 (12.9%) unreported margin status.The 5-year overall survival rate (5yOS) for the 3 528-patient cohort was 23.0%.The 5yOS for patients with resectable tumor was 39.6%, for patients with stage ⅣB tumor without surgery was 5.4%, and for patients with stage ⅣB tumor underwent palliative surgery was 4.7%.Conclusions:More than half GBC patients in China are diagnosed in stage Ⅳ.Curative intent surgery is valuable in improving prognosis of resectable GBC.The treatment of GBC needs further standardization.Effective comprehensive treatment for GBC is in urgent need.

Objective To understand the epidemic characteristics of different virus types of hand, foot and mouth disease (HFMD) in Zhaotong City, and provide guidance and recommendations for the prevention and control of HFMD, and to analyze seasonal characteristics of different virus types of HFMD in Zhaotong City. Methods The epidemiological characteristics of different virus types of HFMD in Zhaotong City from 2014 to 2017 were analyzed using the concentration and circular distribution methods. Results The main pathogens detected were EV71, Cox A16 and other enteroviruses, which were 216, 182, and 294, respectively, accounting for 57.45%, 73.44%, and 67.11%. M was 0.86, indicating that EV71 had strong seasonality. The Rayleigh test showed statistically significant differences (Z = 99.53, P <0.001). ā = 157 °, the peak day of onset was May 10, similar to untyped (May 16), the peak period was April 21-June 1, and the epidemic period was April 1-June 21. Conclusion According to the incubation period of hand-foot-mouth disease and the period of time during which the vaccine develops protective effects, vaccination of hand-foot-mouth disease vaccine at the peak period has a good guiding significance for the timeliness and pertinence of vaccination.