Objective To examine and analyze the methylation status of the promoter region of human telomerase reverse transcriptase(hTERT)in gastric cell lines.Methods Telomerase activity was detected with telomeric repeat amplification protocol(TRAP).Expression of hTERT was observed by immunofluorescence.The target sequence of hTERT CpG island(-692~-403bp)was selected by database search.The methylation status of hTERT CpG island was examined with bisulfite sequencing PCR(BSP)as following:Genomic DNAs were modified by sodium bisulfite and amplified by PCR with primers that contained no CpG sites.Direct sequencing was performed for each case.Results Both TRAP and immunofluorescence analysis showed that high levels of telomerase and hTERT were expressed in SGC-7901 and BGC-823 cells.The sequences(-634~-403bp)of hTERT gene promoter,which were sequenced successfully,were homologous to the sequences before sodium bisulfite modification.25 CpGs in BGC-823 cell line and 26 CpGs in SGC-7901 cell line were observed to be methylated in all 26 CpGs of this part.Only one repressive transcription factor,which called myeloid-specific zinc finger protein 2(MZF-2),was found to bind this region which possessed two special binding motifs.Such a kind of binding showed potential importance for methylation-mediated derepression of hTERT transcription.Conclusion The hypermethylation may change the structure of DNA and result in abortive binding between MZF-2 and its motif,which might be one of the reasons for up-regulation of hTERT transcription and telomerase activity.

Objective To investigate the clinical characteristics and treatment method of pulmonary metastasis of transitional cell carcinoma of urinary tract.Methods The clinical data of 18 patients with pulmonary metastasis of transitional cell carcinoma of the urinary tract,who were admitted to the Second Hospital of Tianjin Medical University from July 2005 to March 2009 were analyzed.Results The primary sites of transitional cell carcinoma of these 18 patients were renal pelvis(8 cases),ureter(6 cases) and bladder(4 cases).Multiple metastases to both lungs were found in 14 cases,the chest X-ray showed dense nodular shadows in different size diffused in both lungs,and CT examination revealed the same picture.All the 14 patients were treated with systemic chemotherapy,and 13 of them underwent surgical operation after the discovery of local recurrence in their bladders.Isolated lung metastasis was found in 4 cases,in whom chest X-ray showed isolated dense nodular shadow in the lung,and CT examination revealed soft tissue shadows adjacent to the chest wall.All the 4 patients were treated with implantation of radioactive pellets and systemic chemotherapy,and one of them underwent surgical operation after the discovery of a local recurrence in the bladder.Follow-up was conducted for 6-36 months.Two patients died in 6-12 months,14 died in 12-24 months,and 2 survived for longer than 24 months.Conclusions The incidence of pulmonary metastasis of transitional cell carcinoma of urinary tract is low.Chest radiograph and CT are the main diagnostic means.Systemic chemotherapy should be used as the main treatment for diffuse pulmonary metastasis,radioactive pellet implantation and systemic chemotherapy should be used for a localized isolated pulmonary metastasis,and surgical operation should be done for the patient with local recurrence in the bladder.

Objective Compared with the effect of thromboembolectomy assisted with intraoperative digital subtraction angiography (DSA) and traditional method,evaluate the value of intraoperative DSA on surgical procedures for acute lower limb arterial embolism. Methods Sixeight cases of traditional thromboembolectomy to take on the operation (group A) and 72 cases under the DSA operation (group B)were analyzed during January 2005 to December 2009, Comparing two sets of operation (time, the amount of bleeding and the ankle brachial index changes before and after the surgery , amputations rate). Results The operation time of group B [(76 ±17) min] was less than that of group A [(95 ± 22) min, t =5. 736, P ＜ 0. 01] ; the amount of bleeding of group B [(83 ± 35 ) ml] was significantly less than group A [(102 ± 58 ) ml, t = 2. 362, P ＜ 0. 05]; The difference of ABI in group B after the operation than before (0. 32 ±0. 08) than that in group A(0. 25 ±0. 12) had remarkably improved ( t =2. 33, P ＜0. 05) ;there were 8 patient amputations in group A and 2 in group B ( u = 2. 06 , P＜ 0. 05 ); there were 2 patients died in group A and one in group B ( P ＞ 0.05 ). Conclusion Thromboembolectomy assisted with DSA can be accurate assessment of the arteries embolism position, elevate the success rate in the operation ,reduce the rate of amputation saws.

Objective:To investigate the effect of potassium sodium hydrogen citrate in the treatment of renal uric acid calculi with heavy load.Methods:The clinical data of 6 patients with intrarenal high-load uric acid calculi (long diameter >4 cm) treated with potassium sodium hydrogen citrate in our hospital from January 2018 to July 2020 were reviewed. All of the patients were male. Their ages ranged from 42 to 66 years, with an average age of 51.3 years old. The average length to diameter was 6.0(4.1-7.6) cm. The average density of stone was 475 (418-535) HU. The average blood uric acid was 453.3(258.7-570.0)μmol/L, and all patients had a urine pH ≤5.5. The blood uric acid level was higher than normal serum uric acid level before treatment, and 3 cases had a history of gout. Stone composition analysis revealed 100% uric acid stone in two patients before treatment. The remaining patients were more likely to have uric acid calculi before treatment. None of the patients had a history of urinary tract infection. All patients were treated with oral potassium sodium hydrogen citrate. During the treatment, the starting dose was 10g/ day, which was divided into 2.5 g after breakfast, 2.5 g after lunch and 5.0 g after dinner. The dose was adjusted according to the pH value of urine, and the urine pH was maintained between 6.5-7.0. CT plain scan was repeated every 2-3 months during the treatment period to evaluate the treatment effect.Result:After 2.5-8.0 months’ treatment, the stone load of all the patients was reduced to different degrees, and the average length diameter was shortened by 3.2cm, among which two patients’ stones disappeared. CT scan showed the stone edge changed from smooth to rough after the treatment, and there would be worm erosion defect and cavity on the surface and inside of the stone, showing obvious stone dissolution phenomenon.Conclusions:Potassium sodium hydrogen citrate has a good therapeutic effect on renal uric acid calculi with heavy load. Non-infected patients with uric acid calculi should be treated with potassium sodium hydrogen citrate.

OBJECTIVE To assess the outcomesof three different therapeutic approaches for the treatment of T3N0M0(stage Ⅲ) glottic squamous cell carcinoma. METHODS Sixty-five cases of T3N0M0 glottic squamous cell carcinoma treated with curative intent by three different modalities include vertical hemilaryngectomy(VHL,n=21),total laryngectomy(TL,n=31)and radiotherapy(RT,n=13) were reviewed retrospectively. The survival rate,recurrence rate at the primary lesion site and jugular lymph node, and laryngeal preservation was compared among three methods. RESULTS There was no significant difference in the overall survival rates(Cox proportion hazard model) and recurrence rates at the primary lesion site or jugular lymph node among the three methods. Laryngeal function was preserved in 100% of the cases in the VHL and RT groups after initial treatment. CONCLUSION The three treatment modalities had statistically similar survival and recurrence rates. Patients treated with VHL and RT had a higher rate of laryngeal preservation compared to that of TL, hence VHL or RT is a valid alternative to TL in treating selected patients with T3N0M0 glottic squamous cell carcinoma.

Objective To investigate the effects of antisense recombinant euraryotic expression vector of HCCR-2 on the proliferation and apoptosis of HepG2. Methods The antisense recombinant eukaryotic expression vector of HCCR-2 was constructed. The vector was stably transfected to the HepG2 cells, and positive clones were selected by G418 (antiseuse vector group), pIRES2-EGFP vector was transfected into the HepG2 cells in the same way (pIRES2-EGFP group). The conditions of the nontransfected HepG2 cells were used as control (HepG2 group). Changes in cell growth curve, cell cycle, cell apoptosis and morphology of HepG2 cells after the transfec-tion were detected by MTT method, flow cytometry and transmission electron microscopy, respectively. All the data were analyzed by one-way ANOVA and chi-square test. Results The expression level of HCCR-2 mRNA was down-regulated to 0.39±0.04 in antisense vector group, and the expression level of HCCR-2 mRNA in pIRES2-EGFP group and HepG2 group were 0.62±0.06 and 0.72±0.03, respectively, with significant difference among the 3 groups (F=43.701, P<0.05). The apoptotic rate of HepG2 cells in antisense vector group, pIRES2-EGFP grop and HepG2 group were 13.30%, 2.51% and 2.07%, respectively, with significant difference among the 3 group (χ2=6.793, 8.721, P<0.05). The growth of HepG2 cells in antisense vector group was retarded, and was blocked in G0/G1 stage. Conclusions The HCCR-2 antisense recombinant eukaryotic expression vector can inhibit the mRNA expression of HCCR-2 and promote the apoptosis of cells. HCCR-2 may be involved in cell regulation and the proliferation of hepatocellular carcinoma cells.

ObjectiveTo study the effect of ischemia postconditioning intervention in a rabbit's acute mesenteric ischemia-reperfusion injury model.Methods 120 rabbits were divided randomly into Con( only expose SMA by operation),I/R( clamping SMA 30 min,reperfusing 120 min),IpostC1 ( clamping SMA 30 min,3 clamping 30 s/releasing 30 s round,reperfusing 117 min),and IpostC2 (clamping SMA 30 min,3 clamping 60 s/releasing 60 s round,reperfusing 114 min) group (n =30).Levels of MDA and MPO in serum and intestinal tissues were measured. Chiu-6 standard scoring was used to determine the pathology score of injured intestinal mucosae.ResultsCompared with the Con group,MDA and MPO levels in serum and intestinal tissues increased obviously in the three other groups,the same as in the pathology score of injured intestinal mucosae (P ＜ 0.01 ) ; Compared with the I/R group,the MDA and MPO levels in serum and intestinal tissues decreased obviously in the IpostC1 group ( P ＜ 0.01 ),but not in the IpostC2 group ( P ＞ 0.05 ).ConclusionsMDA and MPO levels in serum and intestinal tissues and intestinal mucosal injury decreased obviously in the rabbit's acute mesenteric ischemia-reperfusion injury model by ischemia postconditioning intervention.

BACKGROUND: Autism spectrum disorder (ASD) encompasses a range of disorders that are characterized by social and communication deficits and repetitive behaviors. This study evaluated the effect of methyl-6-(phenylethynyl)-pyridine (MPEP), an antagonist of the mGluR5 metabotropic glutamate receptor, on memory enhancement in the BTBR T+tf/J (BTBR) mouse strain, which has been recognized as a model of ASD. METHODS: The pharmacological effects of MPEP on memory and motor coordination were assessed using the Morris water maze and rotarod tests in BTBR and C57BL/6J (B6) mice. Furthermore, we performed morphological analyses of cerebellar foliation in BTBR and B6 mice using hematoxylin and eosin staining. RESULTS: MPEP-treated BTBR mice exhibited improved learning and memory in the Morris water maze test. MPEP administration also improved motor coordination in the rotarod test. However, no significant difference was observed regarding the numbers of Purkinje cells in the cerebella of BTBR versus normal B6 mice. CONCLUSION: This study suggests that the mGluR5 antagonist MPEP has the potential to ameliorate learning and memory dysfunction and impaired motor coordination in BTBR mice. These results further suggest that the BTBR mouse model may be useful in pharmacological studies investigating drugs that could potentially alleviate cognitive dysfunction in ASD.
Animals ; Child ; Autism Spectrum Disorder* ; Eosine Yellowish-(YS) ; Hematoxylin ; Learning ; Maze Learning ; Memory ; Memory, Short-Term* ; Mice* ; Purkinje Cells ; Receptors, Metabotropic Glutamate ; Rotarod Performance Test

Objective To evaluate the feasibility and clinical effect of Argon plasma coagulator in simple enucleation for small renal cell carcinoma. Methods On the basis of successful performing the animal experience of coagulating therapy on the wound tissue during partial nephrectomy with Argon plasma coagulator in rabbit models, 10 cases of simple enucleation for small renal cell carcinoma with Argon plasma coagulator were accomplished. Results Both with the standard of stopping bleeding of wound tissue by Argon plasma coagulator and with the standard of re-spraying the wound tissue for 2 s after stopping bleeding using Argon plasma coagulator, the depth of wound tissue necrosis without blocking the renal pedicle is deeper than that with blocking the renal pedicle(P=0. 012 and P=0. 002, respectively).If the wound tissue was re-sprayed for 2 s after stopping bleeding by Argon plasma coagulator, the depth of the wound tissue necrosis without blocking the renal pedicle was deeper than that just with blocking the renal pedicle(P=0. 007 and P=0. 002,respectively). In the part of application in clinical, all procedures were successfully completed. The mean operative time was 163 min (range, 100-210 min) and mean blood loss was 230 ml (range, 100-400 ml). Drainage tube was pulled out 1 month after operation in 1 case for being allergic to absorbable hemostatic gauze, and the mean pulling drainage tube out time in others was 4. 2 d (range, 3-5 d). During a mean follow-up of 22 months (range, 10-38 months), no local tumor recurrence and distant metastasis was found. Conclusion Argon plasma coagulator can be used in simple enucleation for small renal cell carcinoma, and the clinical effectiveness is ideal.

Objective:We designed and applied a novel, bionic megaprosthesis for distal femur, which substituted the hinged articulation with an artificial ligament and a constrained tibial insert in order to maintain articular stability and reduce stress and the risks of wearing, loosening and breakage.Methods:We reviewed the cases of distal femoral osteosarcoma in children population who were initially treated in our center during 2019. Twelve cases of bionic distal femoral prosthesis (bionic group) and 21 cases of fixed-hinge distal femoral prosthesis (control group) were included. In the bionic group, there were six female and six male with the mean age as 8.8±2.6 y. One patient had pathological fracture. Staging according to Enneking system included 1 case of stage IIA, 10 cases of stage IIB and 1 case of stage III. In the control group, there were seven female and fourteen male with the mean age as 8.6±2.2 y. Pathological fracture was seen in three cases. Staging included 15 cases of stage IIB and 6 cases of stage III.We investigated the peri-operative safety, oncological outcome, complication profiles and post-operative functional status of the bionic prosthesis by comparing the baseline data, operative data, and oncological and prosthetic outcomes between the two groups.Results:The two groups were comparable in terms of baseline data including gender, age, height, weight and onset duration, and operative data including duration of operation (126.7±27.9 min vs 143.3±38.9 min, P=0.203), intra-operative hemorrhage (162.5±212.3 ml vs 247.6±175.6 ml, P=0.224) and duration of wound drainage (6.3±2.4 d vs 6.4±3.4 d, P=0.908). The mean follow-up duration of bionic group was 16.0±4.7 months, during which time three patients had systemic progression and one of them died of disease. Another patient had local recurrence and subluxation of the prosthesis that was treated by amputation. Subluxation might be related to the huge tumor mass and excessive resection of the quadriceps femoris. As for the control group, the mean follow-up duration was 12.7±4.5 months. Three patients had systemic progression and all patients were still alive by last follow-up. Local recurrence was seen in two patients and was treated by excision. No prosthetic complications were seen in the control group. The averaged range of motion (ROM) was greater in the bionic group than that in the control group (120.6°±13.6° vs 92.0°±7.7°, P<0.05), but the MSTS 93 scores were similar between groups (29.1±0.9 vs 29.5±0.6, P=0.337). Conclusion:Compared with the fixed-hinge distal femoral prosthesis, the bionic prosthesis had good peri-operative safety, did not increase the risks of prosthetic complications, and could achieve a better range of motion and a similar functional status based on the results of short-term follow-up.