The MegaBACETM 1000 DNA Sequencing System was made by Amersham Pharmacia,which is a high throughput fluorescence-based DNA sequencer that can be used for DNA sequencing and genetic analysis etc.But the markers(ET550-R Size Standards)produced by this corporation which are used for genetic analysis in this instrument are very expensive.In order to cut the cost,a set of markers were developed and after authentication,that can be used in the MegaBACE 1000 DNA sequencer.Using these markers and our own recognition software,an improved,high throughput AFLP work protocol based on MegaBACETM 1000 were constructed.

Objective To investigate the influence of the eye acupuncture therapy in the expressions of brain derived neurophic factor(BDNF)and tyrosine kinase receptor B(TrkB)in penumbra brain tissue of ischemic area in the rats with acute cerebral ischemia reperfusion inj ury,and to explore the related mechanism of protective effect on brain of eye acupunture.Methods 62 SD rats were randomly divided into blank control group (n=11),sham operation group (n=11)and model copy group (n=40).The rats in copy model group were used to establish models with suture method,and the successful model rats (n=32)were randomly divided into model group(n=16)and eye acupuncture group(n=16).The rats in eye acupuncture group were treated with eye acupuncture intervention;the acupoints were chosen according to human acupoint selection method, the liver, the upper-jiao, the down-jiao district,and renal were selected for acupuncture intervention. After reperfusion for 2 h,the acupuncture was performed once every 8 h,lasted for 10 times;30 min after the last intervention,the rats were sacrificed;the regional ischemia penumbra around the parts of the brain tissue was obtained and the expressions of BDNF and TrkB mRNA and protein in brain tissue of the rats were detected with RT-PCR and Western blotting method. Results Compared with blank control group,the expression levels of BDNF and TrkB mRNA and protein in brain tissue of the rats in sham operation group had no statistical significance(P>0.05);compared with sham operation group,the expression levels of BDNF and TrkB mRNA and protein in brain tissue of the rats in model group were significantly up-regulated(P<0.01);compared with model group,the expression levels of BDNF and TrkB mRNA and protein in brain tissue of the rats in eye acupuncture group were significantly up-regulated(P<0.05 or P<0.01).Conclusion The expressions of BDNF and TrkB are increased after cerebral ischemia and reperfusion injury.Eye acupuncture can protect the brain of MCAO/R rats,which may be related to the up-regulation of the expressions of BDNF and TrkB in the corresponding penumbra.

Objective to observe the influence of Candida albicans infection on t cell subsets in mice with spleen deficiency,so as to explore the immune mechanism. Methods totally 150 healthy SPF mice were randomly divided into four groups:control group(N1 group,n = 30);Candida albicans infection group(N2,n = 40),spleen deficiency model group(M1,n = 40),spleen deficiency model combined with Candida albicans in-fection group(M2,n = 40). N2 and M1 mice were infected by Candida albicans at a concentration of 2×108 CFU/mL. ten mice were randomly se-lected from each group at 7,14,and 21 days after infection respectively,and the index was detected. Flow cytometry was used to detect the percent-age of CD4+/CD8+t cells in intestinal mucosa of mice,and the expression level of IL-4 and IFN-γ mRNA was detected by Rt-PCR assay. the levels of IFN-γ and IL-4 were detected by Western blot assay. Results Compared with N1 group,the proportion of CD4+t cells in the lamina of the natu-ral layer of the small intestine was decreased(P < 0.01),the proportion of CD8+t cells was increased(P < 0.01),the ratio of the two was significant-ly decreased(P < 0.01),and the expression levels of IL-4,mRNA IFN-γ and protein in the small intestine tissues were increased in other groups (P < 0.01). In addition,the expression level of IFN-γ was significantly increased in M2 group,while the expression of IL-4 was significantly in-creased in N2 group. Conclusion the susceptibility of Candida albicans infection was increased in spleen deficiency mice,which may be closely related to the regulation of th1/th2 balance.

Objective To learn the current situation,effect and the matters of the internet class teaching quality assessment system and to propose some suggestions and measures for teaching quality assessment scientifically.Methods We made a questionnaire among the undergraduates of the Chongqing Medical University,using Epidata3.02 for data recording and SAS8.2 for data analysis.Result The accessibility of the class teaching quality net assessment system was not very ideal,and most students were not familiar with the pathway for assessing the teaching quality on internet;the students were generally objective and fair in the teaching quality assessment,but they were not very energetic and initiative;the students had the desire to communicate,but the teachers did not think highly of the teaching assessment;the current assessment index and the internet class teaching quality assessment system should be improved.Conclusion We should emphasize the publication and education of the internet class teaching quality assessment system,raise the recognition of both teachers and students on the role and significance of the internet assessment system,upgrade and improve the assessment index system and promote the close association of the assessment and the actual teaching status.

Objective To study the effect of behavioral therapy combined with Paroxetine in the treatment of overactive bladder accompanied by depression and anxiety.Methods Over the past two years,a total of 69 cases of patients with overactive bladder accompanied by depression and anxiety were diagnosed by outpatient,they were divided into experimental group (n=33)and control group(n=36).The experimental group were given behavior therapy and Paroxetine in the treatment of anxiety,depression,while the control group were given behavior therapy.Then the overactive bladder symptom score(OABSS),urgency score,SDS,SAS score before and after treatment of the two groups of patients were statistically analyzed.Results (1)The OABSS score ((3.30± 1.01) vs (7.51 ± 0.69)),urgency score((2.60±0.51) vs (3.93±0.69)),SDS score((43.1±6.2) vs (66.4±4.7)) and SAS score ((41.9±0.6) vs (61.4±3.9)) decreased after treatment of the experimental group.There were statistically significant compared with before treatment(t=17.8773,8.9045,17.2039,16.0273,all P＜0.01).(2) The OABSS score,urgency score decreased after treatment of the control group.There were statistically significant compared with before treatment (t=6.1926,6.3483;both P＜0.01).SDS,SAS score before and after treatment of the control group were not statistically significant (t=1.3105,0.5852,bothP＞0.05) (3) The OABSS score,urgency score,of the experimental group were more depressed than the control group,which were statistically significant (t =3.3830,3.6391,both P＜0.01).Conclusion For overactive bladder patients with anxiety and depression,behavioral therapy combined with Paroxetine is better than behavioral therapy alone.

2-Keto-L-gulonic acid (2-KLG), the precurcor of L-ascorbic acid synthesis, was prepared directly from D-glucose by tandem fermentation. In the first step fermentation Erwinia sp. SCB 247 translated D-glucose to 2,5-diketo-D-gluconate (2,5-DKG), which accumenlated 180 mg 2,5-DKG per ml in the broth. In the second step fermentation Corynebacterium sp. SCB 3058 reduced 2,5-DKG to 2-KLG, which accumulated 35 mg 2-KLG per ml in the broth. This reductive fermentation was obtained under aerobic conditions by adding a hydrogen donor such as glucose.The average yield of five batches fermentation was 56.3 mol%, from D-glucose to 2-KLG in 10 L fermentor.

Objective To evaluate the clinical diagnosis and treatment of acute renal infarction.Methods Two cases (3 sides) of acute renal infarction were reported.The patients were 1 male and 1 female,with the age of 62 and 54 years.Case 1 presented acute left flank pain,and enhanced CT showed a non-enhanced area in the upper and mid pole of the left kidney.The diagnosis of focal renal infarction was made and treated with low-molecular heparin (6000 U ).Case 2 presented acute both right abdominal and flank pain,and enhanced CT showed right renal artery embolism and right renal complete infarction.Digital subtraction angiography (DSA) and catheter thrombolytic therapy was applied.4 months later,the patient presented acute left flank pain,and enhanced CT showed a low density area in left kidney without enhanced by contrast,and DSA and catheter thrombolytic therapy was applied again.Results In case 1,contrastenhanced MRI showed a still low signal area like enhanced CT after 2 days of treatment.The renal function remained normal in the follow-up of 36 months.In case 2,the right kidney resorted to moderate blood flow but became atrophy later.In the follow-up of 4 months,a recurrent focal infarction was confirmed in left kidney by enhanced CT.The left kidney also resorted to moderate bloodflow after DSA and catheter thrombolytic therapy.The renal function became normal after follow-up of 10 months and no new infarction was observed.Conclusions The diagnosis of acute renal infraction could be made by enhanced CT or MRI.Early diagnosis and location of the infraction renal artery is critical for recovery of the impaired renal function.Acute renal infraction should be suspected in patients with unexplained persistent and steady flank or abdominal pain in emergence department.

Objective:To evaluate the efficacy and safety of drug-coated balloon (DCB) with paclitaxel in the treatment of femoropopliteal artery in-stent restenosis.Methods:From Dec 2016 to Jul 2020, clinical and follow-up data of femoropopliteal artery in-stent restenosis (ISR) treated with paclitaxel DCB were retrospectively analyzed.Results:Firty-two patients (56 lower limbs) with femoropopliteal artery ISR underwent DCB therapy. According to Rutherford classification, 1 case was R2 (1.7%), 9 cases were R3 (23.2%), 23 cases were R4 (41.1%), 15 cases were R5 (26.8%) and 4 cases were R6 (7.1%). According to Tosaka classification of ISR, 46 (81.2%)limbs were Tosaka Ⅱ, 10(17.9%)limbs were Tosaka Ⅲ Mean lesion length of ISR was (240±122)mm. Bail-out stent implantation was performed in 25% cases. The median follow-up time was 18 months. The all-cause mortality rate was 11.8%, the major amputation rate was 5.9%, the primary patency rate was 53.4%, the primary assisted patency rate was 67.1%, the secondary patency rate was 93.2%, and the F-TLR was 77.2%.Conclusion:DCB is a safe and effective endovascular therapy for femoropopliteal artery ISR.

Infectious stones are produced by urease producing microorganisms, which have a fast generation rate, high recurrence and mortality rates, and are prone to complications related to infection. At present, the treatment of infectious stones includes surgical treatment and drug treatment, and the research on its treatment methods has become one of the hotspots in the field of urology. This article provides a review of the research progress in the treatment of infectious stones, with the aim of improving understanding of the treatment of infectious stones.

Acute gallstone pancreatitis (AGP) is a kind of acute pancreatitis caused by gallstones. The etiology of AGP is complex, and the anatomic basis and initiating factors have a synergistic effect on its pathogenesis, which needs to be studied jointly. The way of the confluence of pancreaticobiliary ducts, dilated main pancreatic duct, the relatively narrow opening of duodenal papilla and small stones or microlithiasis may be involved in the pathogenesis of AGP, in which small stones are the most important. Etiological diagnosis and clinical treatment of AGP should be carried out simultaneously. The timely selection of treatment methods for different causes can alleviate the patient's condition to the greatest extent and reduce the cost of treatment. At present, it is difficult to unify the prediction indexes of AGP. Meanwhile, the pathogenesis and related prophylaxis and treatment also need to be studied. In this paper, the anatomic basis, initiation factors, pathogenesis and self-defense of AGP were analyzed to provide a new perspective for its treatment.