ABSTRACT
Radiation-induced lung injury (RILI), one of the common complications caused by radiotherapy, encompasses two phases: an early phase known as radiation pneumonitis (RP) and a late phase called radiation fibrosis (RF), threatening the life and life quality of patients, with poor prognosis. Accumulating evidence has shown that the occurrence of RILI is related to a variety of cytokines and signaling pathways. This paper summarized the research on the effects of Chinese medicine on RILI from the perspective of cytokines and signaling pathways. Cytokines include transforming growth factor-β1 (TGF-β1), interleukins (ILs), tumor necrosis factor-α (TNF-α), platelet-derived growth factor (PDGF), vascular endothelial growth factor (VEGF), and high mobility group box-1 (HMGB1). Related signaling pathways are phosphatidylinositol-3-kinase/protein kinase B(PI3K/Akt) signaling pathway, mitogen-activated protein kinase (MAPK) signaling pathway, Wnt/β-catenin signaling pathway, Notch1/Jagged1 signaling pathway, and nuclear factor-E2-related factor2/antioxidant response element (Nrf2/ARE) signaling pathway. Cytokines may interfere with RILI progression by initiating various downstream signaling pathways, such as TGF-β1/Smads signaling pathway, TGF-β1/VEGF signaling pathway, TNF-α/nuclear factor-κB (NF-κB) signaling pathway, and HMGB1/Toll-like receptor 4 (TLR4) signaling pathway. In recent years, many scholars have attempted to delay RILI progression by down-regulating the expression of cytokines, antagonizing the effect of cytokines or regulating signaling pathways. It has been verified that many Chinese medicines, Chinese medicine monomers, and compound Chinese medicine prescriptions can inhibit the release of some cytokines or regulate some signaling pathways to reduce the incidence/severity of RILI, with satisfactory therapeutic effects, which have attracted the interest of scholars.
ABSTRACT
PLCζ is a new isoenzyme of the PLC family which plays an important role in activating mammalian oocytes. In recent years, large-scale expression and purification of active PLCζ protein in vitro for structural biology research has not been successful. In this study, the recombinant human PLCζ protein was expressed and purified in the baculovirus expression system. First, the full length of human PLCζ gene was cloned into the pFastBac-HTA plasmid to form the recombinant donor plasmid that was further transformed into DH10Bac Escherichia coli cells to construct the recombined bacmid by the site-specific transposition that was screened by resistance and blue-white spots. Then the bacmid was transfected to Sf9 insect cells via cellfectin to package the recombinant baculovirus. After the amplification of the recombinant baculovirous, the recombinant protein was expressed from the cells transduced by the recombinant baculovirus and was purified by Ni-NTA resin. Purified protein was identified by Western blotting and time-of-flight mass spectrometry and the enzyme activity was determined. The results showed that the recombinant PLCζ protein in the Sf9 cells was achieved at 72 hours after baculovirus infection and expressed in secreted form in cell culture medium. The recombinant protein purified by Ni²⁺ affinity column was identified as PLCζ by Western blotting and ionization time-of-flight mass spectrometry and the enzyme activity was up to 326.8 U/mL. The experimental results provide a reference for the large-scale production and biological application of recombinant human PLCζ protein.
Subject(s)
Animals , Humans , Baculoviridae , Genetic Vectors , Recombinant Proteins , Sf9 Cells , SpodopteraABSTRACT
Objective Bladder cancer , which has a high rate of recurrence and invasion , is the most common genitourinary cancer.The article was to study the effect of specific chemokine receptor CXCR 4 on invasion capacity and intraluminal implantation of human bladder cancer cells . Methods A CXCR4 specific recombinant plasmid vector (short hairpin, shRNA) was constructed to select those cells which could inhibit the expression of CXCR 4, and these cells were divided into blank control group , negative control plasmid group and recombinant plasmid group (pshRNA-CXCR4-1, pshRNA-CXCR4-2).RT-PCR and immunofluorescence technique were used to detect the mRNA and protein expression of CXCR 4 respectively .Invasion capability in vitro of the cells was evaluated by Boyden chamber .20 nude mice were randomly divided into experimental group and control group ( n=10 ) .The experimental group was established by injection of 100μL shRNA-EJ-M3 into the bladder , while the control group was established by injection of 100μL EJ-M3, aiming to detect the effect of shRNA-CXCR4 on intraluminal implantation of human bladder cancer cells . Results The CXCR4 mRNA expression of the pshRNA-CXCR4-1 group (62.05 ± 1.35) was significantly lower than that of blank control group (174.38 ±1.96, P 0.05).In immunofluores-cence experiment, the red cell amount of the pshRNA-CXCR4-1 group(32.24 ±2.23) was lower than that of the blank control group (89.61 ±4.47,P0.05).The Boyden chamber experiment showed that the number of penetrating cells of the pshRNA -CXCR4-1 group (39.67 ±8.45) was significantly lower than that of the blank control group (135.33 ±9.28, P<0.05) and that of the negative control plasmid group(123.63 ±6.36, P<0.05).As to the intraluminal implanting capability, the difference between the ex-perimental group and the control group of statistical significance (10%vs 70%,P<0.01). Conclusion CXCR4 shRNA can inhibit the expression of CXCR4 and significantly decrease the invasion capacity and intraluminal implantation of human bladder cancer cells .
ABSTRACT
Clinical bilingual education is an effective method to educate modern senior medical talents.In order to cultivate a group of medical talents with good clinical competence,high professional standard of English,the model of “Bilingual teaching,goal-directed motion and learning by using binding” in combination with modern advanced teaching means is worth discussing.
ABSTRACT
Objective To discuss the long term clinical effect of ileal orthotropic neobladder.Methods From 1991 to 1998,79 patients,mean age 55(41~75)years,male 74,female 6,were followed up.The serum creatinine and urea,electrolytes,blood routine,B ultrasonic scan of the neobladder residual urine and IVU or MRU of the patients were followed up.The max transverse diameter of renal pelvis and the max verticaI/level diameter of neobladder were measured in 5,10 to 14,15 years of postoperative when IVU or MRU.All results of different time were compared by the multiple comparisons.The local or distant cancer recurrence and the complications of the operation Were evaluated. Results Sixty-four cases,58 male,6 famle,were long term followed up:mean time was 167 (range,121~216)months.Seven cases died of other diseases.Seven cases had pelvic recarrence.Two cases had urethral recurrence.Three cases died of tumor metastasis.One case had ureter recurrence.Forty-eight patients were alive more than 10 years.The value of the serum creatinine,urea,electrolytes and bloods routine of the patients were normal after 5,10 to 14 and 15 vears postoperative (P>0.05).The max transverse diameter of the renal pelvis in 5,10 to 14 and 15 years Dostoperative were 14.0 mm,14.1 mm and 13.7 mm,respectively,P>0.05.The max vertical/level diameter of the neobladder in 5,10 to 14,15 years of postoperative were 110.4 mm/90.4 mm,111.5 mm/95.3mm and 127.0 mm/97.0 mm,respectively,P>0.05.The residual urine of 5 cases was more than 50 ml and had not increased during follow up.Eight cases with neobladder stone were cured by the intracavitary lithothrypsis.Two cases with uretheral stricture were cured by the intracavitary therapy.Twelve cases of 14 cases with inguinal hernia were cured by reoperation,2 cases accepted conservative treatment.Only 17 cases had no complication involve of the cancer and the operation. Conclusion The upper urinary tract and neobladder of the ileal orthotopic neobladder could be stable for long time,the cure rate of tumor is satisfactory and the lifetime follow up is necessary.
ABSTRACT
Objective To summarise the methods for a long segmental avulsed inj ury of ureter.Methods Ureteroscopies were performed on three male patients with upper ureteral calculi and one female patient with haematuria from upper urinary tract.Ureteral calculi caused small and middle hy-dronephrosis,obstruction wasn't seen on the upper urinary tract with haematuria.The kidney on the affected side showed normal founction on IVP.Four patients were suffered from a long segmental avulsed inj uries of full-thickness of ureter from ureteroscopy in a continuous epidural anesthesia.The injuries took place in UPJ and upper ureter.The length of avutsed ureters is 20-25 cm,mean length is 24 cm. Results The ureteroscopies were altered to open operations which were ileal ureteral substi-tutions on two patients,being anastomosed the avulsed ureter wrapped by caul on one patient and nc-phrectomy on one patient under general anesthesia in an emergercy.Two ileal ureteral substitutions had reached satisfied results that were no damaged renal functions,no metabolic acidosis,no repeatly U-rinary tract infections and no complains about micturition.One patient has maintained normal renal morphology and function after thirteen years followup,the other formed stricture in the anastomosis of the proximal piece of-ileum to the renal pelvis tWO years after operation,and then improved after ante-grade dilation.The patient with being replaced back and anastomosed the avulsed ureter had compli-cated with a renal atrophy on the affected side three months after the double-J was extracted.The kid-ney showed no function on isotop nephrogram and then was ablated.The last one with nephrectomy and the one before were followed up regularly,the renal functions and blood pressures remain normal.Conclusion Ileal ureteral substitution would be a good choice and has a stable curative effect and provide a good prognosis for treating long segmental avulsed injury of ureter suffered from ureterosco PY when no available urinary tract was utilized for reconstruction.
ABSTRACT
BACKGROUND: A long-term follow-up indicates that orthotopic ileal neobladder can cause acid-base balance and nutritional metabolic disorder. Otherwise, a long mesenterium is necessary to balance the tension of bladder at pelvic cavity and urinary inosculation due to a high position of ileum. On the contrary, sigmoid neobladder is near by urinary canal, and orthotopic sigmoid neobladder as a bladder substitute after radical cystectomy has few effects on acid-base balance of electrolytes, nutritional metabolism and secretion of mucus.OBJECTIVE: To compare the clinical results of these two operations basis on long-term follow-up. DESIGN: Retrospective analysis.SETTING: Department of Urinary Surgery, the Second Affiliated Hospital of Kunming Medical College.PARTICIPANTS: 164 patients with carcinoma of bladder were selected from Department of Urinary Surgery, the Second Affiliated Hospital of Kunming Medical College form January 1995 to March 2005. Ninety-six of them, including 74 males and 22 females, with age of 43-74 years and the average age of 65 years, accepted the operation of orthotopic ileal neobladder were regarded as the ileal neobladder group, and the other 68, including 64 males and 4 females, with age of 51-72 years and the average age of 62 years, accepted the operation of orthotopic sigmoid neobladder were regarded as the sigmoid neobladder group. All patients were finally diagnosed as pathological examination, and informed consent was provided by all patients. Treatment plan was approved by the local ethical committee.METHODS: ① Orthotopic ileal neobladder: Once the bladder was removed, a segment of ileum about 40-60 cm in length was isolated. In the operative procedure, the distal part of ileum which connected to the caecum often kept, the length of which was 15-20 cm. Both distal ends of the ureters were anastomosed to the homolateral not been split end of the isolated bowel. A perforation was constructed at the bottom of the pouch which served as the outlet, this outlet was then anastomosed to the proximal portion of the remaining urethra. ② Orthotopic sigmoid neobladder: After surgically removing the bladder, a part of the sigmoid colon, the length of which was 30-40 cm was isolated. Other operations were as the same as those mentioned above. MAIN OUTCOME MEASURES: Time of operation, blood loss during the procedure, length of time confined to bed, time of indwelling catheter, the ability to maintain continence and urinate, the results of urodynamic studies, and pouch related complications after operation.RESULTS: In 164 patients, 12 (7.3%) were lost to follow-up. The mean follow-up times were 46 months in the group of orthotopic ileal neobladder and 42 months in the group of orthotopic sigmoid neobladder, respectively. Blood loss during the procedure and the ability to maintain continence and urinate were similar in the two groups (P > 0.05). Compared with sigmoid neobladder group, the ileal neobladder group spent more time on operation, keeping the bed and indwelling catheter. The max volume of ileal pouch was higher than that of sigmoid pouch, and the difference was significant in statistic analysis (t=2.56-3.08, P < 0.05-0.01). Incidence of complication of ileal pouch (16.7%, 29.2%) was higher than that of sigmoid pouch (9%, 16%). The incidence in the early phase was not significantly different, but that in the late phase was significantly different (x2=5.426, P < 0.05).CONCLUSION: Compared with orthotopic ileal neobladder, sigmoid neobladder is worthy of being preferred for its shorter operative time, faster recovery and lower rate of pouch related complications.
ABSTRACT
0.05).Conclusion: The method of constructing a highly invasive subline of bladder neoplasm is reliable,and EGFP transfection does not change the biological behavior of the cells.The obtained sublines may provide a valuable experimental platform for further study on the molecular mechanisms of bladder neoplasm metastasis.
ABSTRACT
<p><b>OBJECTIVE</b>To survey the application of PCR for screening HCV RNA from blood donations within the window period.</p><p><b>METHODS</b>According to a standardized method, 12 blood banks organized by the National Center for Clinical Laboratories collected and prepared about ten thousands specimens. The specimens were tested with two different kits.</p><p><b>RESULTS</b>Among the 7173 specimens A group, 21 were PCR positive for HCV RNA. The positive rate was 0.29%. There were not positive for HCV RNA among 7477 specimens (B group).</p><p><b>CONCLUSIONS</b>It is feasible to use the PCR screening for the detection of HCV RNA of blood donations but is unnecessary to standardize the specimen collection and the kit selection.</p>
Subject(s)
Humans , Blood Donors , Blood Transfusion , Hepacivirus , Genetics , Hepatitis C , Mass Screening , Polymerase Chain Reaction , RNA, ViralABSTRACT
0.05).The operation was failed in 2 cases due to the displacement of the renal tube(re-puncture was performed) or massive hemorrhage from torn renal calices(the procedure was terminated,and the bleeding was controlled by compression and hemostat).One patient developed acute pulmonary edema after the operation and was cured by diuretic;4 cases showed postoperative fever;no serious complications occurred in this series.The stone-free rate in one session was 89.1%(49/55).The residual calculi were all cleared by a second operation combined with extracorporeal shock-wave lithotripsy.The patients were followed up for 3 to 12 months(mean,6 months),during which no recurrent case was found.ConclusionUreteroscopy is safe and effective for establishing renal channel in minimally invasive percutaneous nephrolithotomy.