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.

Objective To evaluate the therapeutic efficacy of hook plate and fascia lata graft in management of chronic acromioclavicular joint dislocation.Methods From May 2012 to March 2014,a total of 16 cases of chronic acromioclavicular joint dislocation (Rockwood type Ⅲ) were treated with hook plate and fascia lata graft.They were 10 men and 6 women,from 19 to 57 years of age (mean,34.7 years).Seven cases were at the left side and 9 at the right side.All injuries resulted from direct violence.The time from injury to surgery ranged from 8 to 54 weeks (mean,12.4 weeks).At the final follow-ups,the shoulder function was assessed using the Quick Version of the Disabilities of the Arm,Shoulder and Hand Score (DASH) and visual analogue scale (VAS),and the efficacy using Karlsson's evaluation system.Results All surgical wounds primarily healed.The dislocation of acromioclavicular joint was corrected,without any vascular or neural damage.The patients were followed up for an average of 12 months (from 6 to 18 months).No plate breakage,screw loosening or redislocation was observed.The DASH and VAS scores of the shoulder at the final follow-ups (25.42 ± 8.34 and 2.22 ± 1.70,respectively) were significantly improved,compared with the preoperative ones (43.72 ± 12.35 and 6.33 ± 1.37,respectively) (P ＜ 0.05).According to the Karlsson's criteria,10 cases were excellent,4 good and 2 moderate.Conclusion In management of Rockwood type Ⅲ chronic acromioclavicular joint dislocation,hook plate and fascia lata graft can restore dynamic balance and stability of the acromioclavicular joint,resulting in solid fixation,early joint motion and limited complications.

Objective To investigate the effect of diuretic (furosemide) therapy on kidney injury induced by melamine and cyanuric acid in rats. Methods 36 male Spragne Dawley rats were random disided into 3 groups. Group A was treated with 2mL of water daily, group B was treated with melamine and cyanuric acid ( each 100 mg/kg) daily for 4 days and then 2ml of water daily, group C was treated with the same as group B at the first 4 days and then treatment with furosemide (20mg/kg) daily. Samples of blood and 24h urine were collected to detective biochemical indexes, and kidney sections were performed on days 4 and 11 ( each end point, n = 6). The kidneys were observed with histopathology and renal crystal deposition scores were determined. Results On the 4th day, group B and group C were resulted in acute kidney injury such as oliguria [ ( 3. 39 ± 1.02 ) ml, ( 3. 20 ± 0. 86 ) ml ] and high serum creatinine [ ( 153.54 ±27. 08)μmol/L, (160. 11 ± 19. 55)μmol/L] and renal melamine cyanurate crystal were found in the renal tissues. On the 11th day, the renal crystal deposition score in the rats was reduced by 9. 52% ( P >0. 05). Compared with those of the 4th day in group B, it reduced by 63.63%( P <0.05) in group C. Urine volume were increased significantly compared with those of the 4th day( P < 0. 05 ) in group C [ from (3.20±0. 86)ml to (25.96 ±5.97)ml] and group B [ from(3. 39 ± 1.02)ml to (8. 57 ± 1.66)ml] , and Urine volume in group C was increased significantly more than that in group B ( P < 0. 05 ). The serum creatinine was obviously reduced as compared with those of the 4th day in group B and C( P <0.05), from[ (153. 54±27.08) μmol/L] to [ ( 106. 10 ±5.53) μmol/L] in group B and from [ ( 160. 11 ± 19. 55) μmol/L] to [ (67. 17 ± 12. 80 ) μmol/L] in group C, but the serum creatinine in group B was still higher than that in group A and C ( P < 0. 05). Conclusions Furosemide can attenuate the damage of acute kidney injury induced by melamine and cyanuric acid.

Objective To explore the curative effects of unilateral Orthofix external fixation in the treatment of tibial defects.Methods From June 2012 to February 2016,we treated 19 patients with tibial defects using unilateral Orthofix external fixation.They were 14 men and 5 women,aged from 26 to 79 years (mean,43.0 years).The right tibia was involved in 12 cases and the left tibia in 7.The soft tissue defects averaged 6.4 cm × 3.7 cm.The mean bone defect before bone transport was 8.9 cm in length.The anteroposterior and lateral X-ray films of the affected tibia were taken regularly after surgery.Edwards tibial fracture scores were used to evaluate the functional recovery and complications of the affected limb at final follow-ups.Results All the patients were followed up for 10 to 40 months (average,18.5 months).Bone union time ranged from 4 to 16 months (average,9.0 months).Lengths of bone transport ranged from 6 to 10 cm (average,7.8 cm).All the bone defects were reconstructed.According to the Edwards scoring at the final follow-ups,10 cases were rated as excellent,6 as good and 3 as poor.Complications were observed in 4 cases,giving a complication rate of 21.1％.Conclusion Unilateral Orthofix external fixation and bone transport can lead to preferable curative effects in the management of tibial defects.

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