Objective To investigate the effects of the HyTK gene transduction on murine bladder carcinoma cells in vitro . Methods The retroviral vector plasmid PL(HyTK)SN was transfected into packaging cell line PA317 using the liposome method. The obtained recombinant retroviral supernatant fluid was used to infect murine bladder cancer cell line T739 cells. B resistant colonies, named as T739 TK, were obtained after hygromycin B selection. The integration and expression of HyTK gene in T739 TK cells were identified by RT PCR. The in vitro killing effects of GCV on T739 TK cells were observed. Results RT PCR showed that HyTK gene was inserted into T739 cells successfully and there was mRNA expression of HyTK gene. In vitro , T739 TK cells expressed the killing effects after treatment with GCV of different concentrations. However, T739 cells grew well at different doses of GCV. Conclusion GCV has obvious killing effect on HyTK gene transfected T739 cells in vitro .

Objective To determine the bystander effect of therapy with herpes simplex virus thymidine kinase ( HSVtk ) gene combined with ganciclovir (GCV) on mouse bladder cancer. Methods Mouse bladder cancer cell line (T739) was transfected with retroviral vector HSVtk gene. The sensitivity of T739TK cells to GCV was detected in vitro . Bystander effect in vitro in co cultured mixtures of T739TK and T739 cells at different ratios or at the same ratio in culture bottles of different size was determined. In the mouse model of bladder tumor, mixtures of T739 and T739TK cells were implanted beneath the peritonea of syngeneic mice. When tumors grew to the size of 0 5-0 8 cm, intraperitoneal administration of GCV was carried out for 6 d. Changes of tumor size and survival rate of mice were observed. Results T739 cells retrovirally transfected with the HSVtk gene became sensitive to low concentrations of GCV. Analysis by RT PCR confirmed HSVtk expression in the transfected T739TK cells. An absolute bystander effect was observed in the mixed culture of cells in vitro when the HSVtk gene transferred cells were at the ratio of above 10%. The tumor growth in the animal model was significantly inhibited by GCV, demonstrating the existence of bystander effect in vivo . The survival time of mice was prolonged after administration of GCV. Conclusion The in vitro and in vivo bystander effects exist in mouse bladder cancer transfected with HSVtk gene, enhancing the role of HSVtk /GCV system for the treatment of bladder carcinoma.

Objective:To explore the application effect of optimizing the anti-epileptic drug (AED) administration nursing procedure for patients with severe epilepsy at the first time after ICU admission.Methods:A quasi-experiment study was conducted by convenience sampling. The 44 patients with severe status epilepticus admitted in our department from January 2016 to December 2017 were selected as the control group .The 44 patients with status epilepticus admitted in our department from January 2018 to December 2019 were selected as the experimental group .The experimental group was implemented the optimized administration procedure after ICU admission. The control group was implemented the routine administration procedure after ICU admission. Some data of the two groups were compared, including baseline data, first administration time after admission, administration step time, AED dose reserve, and seizures at different time periods and so on.Results:Comparative analysis of the time of the first drug administration after admission into the two groups .The administration time of the control group was 31.0 (10.0-69.0) min, which was significantly longer than that of the experimental group, 25.0 (16.0-31.8) min ( χ2 value was -2.760, P<0.05). Comparison and analysis of the time taken for each step of drug administration: The time taken for step 1(Patient admission—medical order confirmation) in the control group was significantly greater than that in the experimental group ( χ2 value was -2.811, P<0.05); step 2 (medical order confirmation—Prepare medicines)in the control group was significantly greater than that in the experimental group ( χ2 value was -4.327, P<0.05).There was no significant difference in the time taken for step 3(Prepare medicines—Drug delivery to complete)between the two groups ( χ2 value was -1.137, P>0.05). Control group AED reserve ratio 63.6% (28/44), experimental group AED reserve was 100.0% (44/44), and the difference was statistically significant ( χ2 value was 19.556, P<0.05). Epilepsy control in two groups of patients: the total control rate after the first administration of the experimental group was 93.2% (41/44), which was significantly higher than 61.4% (27/44) of the control group ( Z value was -3.445, P<0.001). Conclusion:It can shorten the time to first give AED and improve the patient's clinical symptoms that optimizing the first drug administration procedure for patients with status epilepticus after admission.

Objeetlve To evaluate the relationship between temporomandibular joint(TMJ)osteoarthrosis (OA)and cytokines.Methods 20 rabbits were enrolled in this study.They were divided into either experiment(n=10)or control group(n=10).The modeh of TMJ osteoarthrosis were formed by injection SMT into upper joint compartment of the TMJ.Cytokines in the aspirated synovial fluid,induding interleukin(IL-1),IL-6,tumor necrosis faetor(TNF-α),were tested before and after SMT iniection.Results The level of IL-1β,IL-6 and TNF-α in experiment group decreased significantly after treatment,there was significance(P＜0.05);and the level of IL-1β,IL-6 and TNF-α in control group is lower after treatment,there was no significance(P＞0.05).Conclusion The cytokines,including IL-1β,IL-6,TNF-α,decreased significantly by injection SMT into upper joint compartment of the TMJ with OA.

Objective To investigate the application valve of bFGF to improve the viability of trans-plantation flap in Wistar rat. Methods Forty-eight Wistar rats were randomly divided into two groups based on the age: group A,B(1.5 month old, every group was 12 rats) and group C,D (3 month old, every group was 12 rats). After an ischemic model completed, recombinant bovine basic fibroblast growth factor (rb-bFGF) was given to groups A and C in ischemie zone by vascularization flap injection and the equality of normal saline to groups B and D. 14 days postoperatively, the muscular tissue was sent for histology, the blood vessel density was calculated by image analysis, and positive VEGF was detected by immunohistochem-istry. Results The member of capillaries and positive VEGF were more in group A than that in B, and also were morein group C than D(P < 0.05), but there were not statistic difference between group A and C (P > 0.05). Conclusion Recombinant bovine bFGF can stimulate angiogenesis and improve the ischemie vascu-larization flap of rat, which is not associated with their age.

Objective To discuss the clinical therapeutic efficacy of the clinical study of thumb tip defect reconstruction with free distal second toe in emergency. Methods Fifteen cases of thumb tip defect were reconstructed with free distal second toe in emergency.The blood supply was restored by anastomosis of digital and toe'S artery and nerves,the dorsal surface and other side vein of fingers all can be anastomosed with toe's vein.but the first was main. Results All of fifteen fingers survived.Followed-up from 3 months to 3 years,all reconstruction thumbs has a satisfying outward appearance,normal nail and fingerprint,well pinch and hold functions, none effect to the foot. Conclusion Repair of thumb tip defects using free distal second toe in emergency enables favorable functional recovery and satisfying outward appearance,the method is simple and rats recover quickly.

Objective:To investigate the preventive effect of different intermittent pneumatic compressiondevices (IPC) schemes on deep vein thrombosis (DVT) of lower extremity in patients with status epilepticus (SE), to provide nursing basis for rational application of IPC in early admission of SE patients.Methods:A similar experimental study was conducted and 76 SE patients in the intensive care unit of the Department of Neurology, Xuanwu Hospital, Capital Medical University from January 2018 to May 2022 were selected by convenience sampling method. The 35 SE patients hospitalized from January 2018 to February 2020 were included in the control group, and 41 patients hospitalized from March 2020 to May 2022 were included in the intervention group.Both groups were given anti-seizure medications (ASMs) to actively control seizures after admission.IPC application frequency and duration in control group were twice a day, 1 hour each time; the frequency of IPC application in the intervention group was once a day for greater than or equal to 12 hours each time.The incidence of lower limb DVT, epileptic seizure, abnormal electroencephalogram discharge and coagulation index related to lower limb DVT formation were compared between the two groups.Results:There was no significant difference in the indexes of fibrinogen, D-dimer and thrombin time between the two groups of SE patients at admission ( P>0.05). At discharge, the indexes of fibrinogen, D-dimer and thrombin time were (4.02 ± 1.71) g/L, 7.06(5.33, 15.01) mg/L and 10.22(7.53, 14.42) s in the intervention group respectively, which were lower than (4.98 ± 1.62) g/L, 11.92(5.75, 15.26) mg/L and 17.96(13.21, 28.14) s in the control group, the differences were statistically significant ( t = 3.03, Z = 4.32, 1.56, all P<0.05). The DVT incidence rate was 62.86% (22/35) in the control group, and the intervention group was 12.20% (5/41). The difference was statistically significant ( χ2 = 21.59, P<0.01). Conclusions:On the basis of effective control of clinical seizures in SE patients treated with ASMs, early IPC use and daily treatment time at least 12 hours can avoid induced seizures, reduce the formation and risk of lower extremity DVT, and improve blood hypercoagulability.

OBJECTIVETo identify pathogenic mutations of TSC1 and TSC2 genes in two familial and one sporadic cases with tuberous sclerosis complex (TSC).

METHODSFor five patients and their family members, potential mutations of the TSC1 and TSC2 genes were detected by direct sequencing.

RESULTSFor one family, a novel missense mutation c.1964C>T (p.S655F) was detected in the exon 19 of the TSC2 gene. For the sporadic patient, a repeat substitution with deletion mutation c.5238-5255delCATCAAGCGGCTCCGCCA (p.His1746GlnfsX56) was detected in the exon 40 of the TSC2 gene, which led to a stop codon TGA after the 56th amino acids. No mutation was found in another family.

CONCLUSIONThe missense mutation c.1964C>T(P.S655F) and the substitution with deletion mutation 5238-5255delCATCAAGCGGCTCCGCCA(p.His1746GlnfsX56) of the TSC2 gene probably underlie the disease in the first family and the sporadic case.

Adolescent ; Adult ; Base Sequence ; Child, Preschool ; DNA Mutational Analysis ; Female ; Humans ; Male ; Mutation, Missense ; Pedigree ; Phenotype ; Tuberous Sclerosis ; genetics ; Tumor Suppressor Proteins ; genetics

OBJECTIVETo identify pathogenic mutation of the TSC1 and TSC2 genes in a patient with tuberous sclerosis.

METHODSPeripheral venous blood samples and clinical data of a pregnant woman with tuberous sclerosis and 4 family members (parents, uncle and husband) were collected. Genomic DNA was extracted. All coding exons of the TSC1 and TSC2 genes and their flanking intronic sequences were amplified by polymerase chain reaction and subjected to direct sequencing.

RESULTSThe patient has presented facial angiofibroma and prefrons fibrous plaque for 20 years, and lumbar connective tissue nevus for 10 years. She also had mental retardation but no epilepsy. A novel frame-shift mutation c.4258-4261delTCAG was detected in exon 34 of the TSC2 gene, which had led to a premature stop codon TAG after the 55th amino acids. The same mutation was not found in the unaffected family members and 100 unrelated healthy controls.

CONCLUSIONThe novel frame-shifting mutation c.4258-4261delTCAG (p.Ser1420GlyfsX55) in the TSC2 gene may be responsible for the disease in the patient.

Adult ; Asian Continental Ancestry Group ; genetics ; Base Sequence ; China ; DNA Mutational Analysis ; Female ; Humans ; Male ; Molecular Sequence Data ; Mutation ; Pedigree ; Pregnancy ; Tuberous Sclerosis ; genetics ; Tumor Suppressor Proteins ; genetics ; Young Adult

Objective To explore the clinical features and risk factors of poor prognosis in neonatal necrotizing enterocolitis(NEC).Methods A retrospective study was carried out in the infants with NEC admitted to 6 cooperative hospitals in Guangdong Province between January 2005 and December 2014.The clinical features and risk factors of poor prognosis in preterm and full-term infants diagnosed NEC,early onset and late onset NEC were analyzed.Results A total of 449 cases who met the criteria were admitted during the study time.The mortality was 23.6％ (106/449 cases),of which the preterm group was 24.6％ (58/238 cases) while the full-term group was 22.7％ (48/211 cases),the early onset group was 22.1％ (45/204 cases) while the late onset group was 24.3％ (57/235 cases).The median number of NEC onset in preterm group was 11 d after birth while the number of the full-term group was 6 d.Full-term infants who diagnosed NEC were more likely to manifest themselves as abdominal distension (52.1％ vs.42.0％,x2 =4.597,P =0.032),vomiting(36.5％ vs.17.2％,x2 =21.428,P =0.000) and bloody stool(30.3％ vs.21.4％,x2 =4.653,P =0.031);but in the onset of NEC,preterm infants more likely to have feeding intolerance (21.0％ vs.12.8％,x2=5.309,P =0.021).The early onset group of full-term NEC was much common in twins or multiplets(9.4％ vs.1.1％,x2 =6.226,P =0.013),which rate of surgical therapy was much higher (41.0％ vs.27.0％,P =0.036) and the breast-feeding rate before NEC was lower than the late onset group(14.5％ vs.32.6％,x2 =9.500,P =0.002),the differences were statistically significant.The gestational age and birth weight were bigger in the early onset group of preterm NEC[(33.8 ±2.5) weeks vs.(32.2 ±2.8) weeks,t =4.261,P =0.000;(2.1 ±0.5) kg vs.(1.7 ± 0.5) kg,t =4.735,P =0.000)],but length of stay was shorter than the late onset group (18.0 d vs.26.5 d,P =0.000).Logistic regression analysis showed that the risk factors of poor prognosis of full-term NEC were shock,peritonitis and sepsis;while risk factors of poor prognosis of preterm NEC were small for gestational age infant,pulmonary hemorrhage,shock,intestinal perforation and sepsis;the risk factors of poor prognosis of the early onset group of full-term NEC was shock;while those of the late onset group were shock and peritonitis;the risk factors of poor prognosis in the early onset group of preterm NEC were shock and sepsis,while those in the late onset group were pulmonary hemorrhage,shock,intestinal perforation and sepsis.Conclusions Compared to the preterm NEC,the onset time of full-term NEC was earlier and the clinical manifestations were more typical.Early identification and management of shock,peritonitis,intestinal perforation,sepsis and pulmonary hemorrhage can reduce the risk of poor prognosis of neonate NEC.