Dermatology ; Humans ; Integrative Medicine ; Physicians

Objective To explore the experiment condition and method for the application of in vitro in vasive Transwell chamber and to observe muscarinicreceptor stimulant and muscarinicreceptor antagonist's influence to cholangiocarcinoma's invasiveness.Methods Two hundred microliter cell suspension of various concentrations(0.5×105/mL,1.0×105/mL,1.5×105/mL and 2.0×105/mL)was added into the upper chamber of the Transwell chamber,and the cells were allowed to penetrate the matrigel for 12,18,24and 48 hours respectively.The numbers was gotten as the invasive cells on the under surface of the membrane.After optimal cell concentration and time were gotten,pilocarpine of various concentrations(0 mmol/L,0.1 mmol/L,0.3 mmol/L and 0.5 mmoL/L)was added into the upper chamber of the Transwell chamber,then the cells on the matrigel were stained and counted.So did the cells when atropine of various concentrations(0.01 mmol/L,0.01 mmol/L,0.05 mmoVL and 0.1 mmol/L)were added into the upper chamher of the Transwell chamber in according to pilocarpine of various concentrations(0 mmol/L,0.3 mmol/L,0.3 mmol/L and 0.3mmol/L).Results With the increase of the time and cell concentrations，the cells couts that penetrated the matrigel increased，while the increase tended to he stable when the culture time exceeded 36 hous and the cell concentration Was over 1.0×105/mL.By adding pilocarpine,there were significant differences between the control and experimental groups(P＜0.05)，but there were no significant differences in experimental groups with various concentrations.There were no significant differences in blank group and experimental groups with atropine added(P＞0.05).When added pilocarpine and atropine,there were significant differences between blank and experimental groups(P＜0.05)，but there were no significant differences in experimental groups with various concentrations.Conclusions Thirty-six hours as invasive time,and one cell concentration 1.0 × 105/mL were optimal to test invasion abilities of cholangiocarcingma cells to different medicines or reagents.There is the possibility that museariniereceptor exists in cholangiocarcinoma cells，and may play an important role in cholangiocarcinoma's invasiveness and metastasis.

Lactic acid production of twelve strains of LAB isolated from broiler intestine and tolerance property of three strains were investigated. The results of lactic acid production showed that among all strains K6 exhibited the most rapid production during the first twelve hours, the seconds were K9 and C1; D17 exhibited the highest production of lactic acid by twenty-four hours, C1 exhibited the highest production of lactic acid by forty-eight hours. The pH values in three strains of K9、D17 and C1 culture showed the fast decline during the first twelve hours, with the final values significantly lower than those of other strains cultures. The results of tolerance property showed that the survival counts of C1could be detected when pH value was at 2 after three hours, but the survival counts of D17 and K9 could not be detected after one hour. When pH value was at 2.5 after three hours ,the survival counts of C1 declined from 10~ 8.2 /mL to 10~ 4.8 /mL, K9 from 10~ 8.2 /mL to 10~ 4.6 /mL, the survival counts of D17 could not be detected. 0.08% bile had few effects on the survival counts of three strains; when incubated in the medium with 0.40% bile, the survival counts of C1 declined from 10~ 8.4 /mL to 10~ 6.5 /mL,D17 from 10~ 10.3 /mL to 10~ 7.5 /mL, and K9 from 10~ 9.8 /mL to 10~ 7.7 /mL. When the group treated with 37℃ for 20 minutes was served as the control, the survival counts of C1 and K9 was not detected when treated with 80℃, but the survival counts of D17 were 10~ 4.9 /mL, when treatment with 65℃ the survival counts of C1 and K9 decreased significantly .

Objective To evaluate the clinical effect and safety of retroperitoneal laparoscopic partial nephrectomy in treatment of patients with T1b renal carcinoma.Methods Fourteen patients (11 males and 3 females) with T1b renal carcinoma were retrospectively performed.The age of patients was (54.5 ± 9.2)years old,with 8 cases on the left side and 6 cases on the right side.Tumor diameter was (5.1±1.3) cm.All the patients received retroperitoneal laparoscopic partial nephrectomy.Results None of the 14 cases was converted to open surgery.The operation time was (112.0 ± 24.7) min,the intraoperative blood loss was (64.6 ± 15.9) ml,the warm ischemia time was (26.5 ± 9.3) min.The 14 patients were not blood transfusion in intraoperative and postoperative.Postoperative negative pressure drainage placement time was (3.1 ± 1.5)d,lying in bed time was 72 h.Serum creatinine increase was found in 1 case postoperative 12 h,others were no severe complications.Postoperative pathology:the incisal margin of 14 cases were all negative,clear cell carcinoma was in 13 cases,the pathology stage was T1bNoM0;angiomyolipoma of kidney was in 1 case.All the patients were follow-up 3-16 (21.4 ± 9.6) months,all the patients had normal renal function and had no tumor recurrence or metastasis.Conclusion Retroperitoneal laparoscopic partial nephrectomy is safe and reliable for treatment of patients with T1b renal carcinoma.

The mammalian cell nucleus is highly structured and organized into various membrane-less nuclear compartments called nuclear bodies. Nuclear bodies are highly dynamic structures, with a variety of substances gathered inside to promote the more efficient conduct of certain biological reactions. It dynamically produces responses under different biological processes and stress conditions such as tumorigenesis, apoptosis, antiviral defense, and plays an important role in regulating cell homeostasis. Tumor is a major public health problem, and finding new targets is the key to tumor therapy. How the nuclear bodies are involved in the development of tumor has not been reported. This review aims to provide a new understanding of how the nuclear bodies regulates tumor progression and provide a new effective strategy for tumor prevention and treatment.

Animals ; Decompression ; Decompression Sickness ; physiopathology ; Electrocardiography ; Electroencephalography ; Electromyography ; Electrophysiology ; Male ; Rabbits

AIM: To clone EBV-LMP2A gene, construct and identify the recombinant retroviral vector and stable cell strains expressing EBV LMP2A. METHODS: The full-length EBV LMP2A gene was generated by RT-PCR amplification from B95.8 cells which contain complement nucleotide sequence of EBV LMP2A gene. The gene was ligated to T-vector and sequenced to construct retroviral vector consisting with LMP2A. To produce retroviral virus, packing cells, 293T cells were co-transfected with recombinant retroviral expression vector pGEZ-LMP2A and two auxiliary viral vectors pHIT456 and pHIT60 by lipofectAMINE2000. Viral titration was performed according to the instructions of the manufacturer. To establish L929 cell line stable expressing LMP2A, L929 cells were infected with recombinant retrovirus three times and selected by Zeocine. The Zeocine-resistant clones (L929/LMP2A) were screened for LMP2A expression by RT-PCR and Western blot. RESULTS: The recombinant retrovirus vector carrying LMP2A gene was constructed successfully. Transfection yield a titer of 5×10~8 infectious particles/L. The infected L929 cells were selected by Zeocine. Results of RT-PCR and Western blot indicated that L929 transgenetic cells could stably express EBV-LMP2A. CONCLUSION: The L929 cell line stably expressing LMP2A provides suitability for extraction of the LMP2A protein and preparations of the vaccine for the therapy of EBV-associated diseases.

Objective To study the pharmacokinetics and detection window of clozapine and its metabolites in human blood, so as to provide experimental basis for forensic cases of identification of clozapine poisoning. Methods 29 Taiyuan Han people's elbow venous blood was collected after given oral administration of 12.5mg clozapine at different time point, in which clozapine and its metabolites were extracted with solid phase extraction (SPE) and determined by HPLC-MS-MS. The qualitative analysis was based on retention time and MRM ions. The quantitative analysis was based on an internal standard method and calibration curve. Using the 3p97 pharmacokinetic software, pharmacokinetic equation of clozapine in the blood were imitated from the C-T data, and pharmacokinetic parameters were calculated. Results The pharmacokinetics of clozapine met a two compartment open model with a first kinetics absorption. The Tmax of clozapine(CLP), demethylclozapine(DMCLP), N-oxidation-clozapine(NO-CLP) respectively were 2.96±1.32h, 8.65±3.00h, 9.31±26.38h; The Cmax of CLP, DMCLP, NO-CLP respectively were 34.68±9.32ng/mL, 11.16±4.15ng/mL, 9.62±13.88ng/mL;The t1/2 of CLP, DMCLP, NO-CLP respectively were 17.02±23.63h, 27.06±12.58h, 41.27±29.75h; The detection window of CLP, DMCLP, NO-CLP respectively were 81.72±26.19h, 93.21±29.40h and 19.93±14.62h. Conclusion The pharmacokinetics of clozapine in blood of Han people is consistent with two compartment open model with a first kinetics absorption. The pharmacokinetics model and parameters of clozapine can provide expirimental basis for forensic identification of clozapine poisoning cases.

Comprehensive biochemistry experiment,which is interlocked and has difficulties in a certain degree,requires considerable knowledge,multiple techniques and long time.In order to ensure the smooth progress of the experiment,biochemistry and molecular biology department of Hebei medical university has taken following measures in teaching preparation and teaching implementation:building a specialized laboratory;performing collective lesson preparation and pre-experiments;technical teaching;teaching with multimedia equipments;students submitting experimental preparatory reports before class and then completing the experiments in groups.These measures achieved the intended purpose of setting up a comprehensive experiment.

ObjectiveTo study the effect of α7 ( α7 AChR) agonist nicotine on regulating sensitivity of regular chemotherapeutic agent in cholangiocarcinoma cells,and explore the possible target.MethodsThe effect of nicotine and α-BTX pretreatment on the survival ability of cholangiocarcinoma cells was investigated when applied with 5-FU by using MTT and Flat cloning formation experiment.ResultsApplied with 5-FU,in various con centrations nicotine stimulating group( 10-3 g/L,10-4 g/L,10-5 g/L ),the survive rate of QBC939 was 128％,124％,118％,while that in α-BTX stimulating group and combined stimulation group was 92％,94％,93％,92％,respectively.The cloning formation ability of nicotine- stimulating group (6.2 ± 0.40) was significantly higher than α- BTX stimulating group (3.2 ± 0.20 ),combined stimulation group ( 3.2 ± 0.20 ) and control group ( 3.4 ±0.33).ConclusionNicotine can prevent chemotherapy-induced apoptosis,and improve cholangiocarcinoma cell survival via α7 nicotine acetylcholine receptor in vitro.