AIM:To investigate the integrative treatment of both coenzyme Q 10 ( CoQ10 ) and minocycline in the rats intranigrally intoxicated with 1-methyl-4-phenylpyridinium ( MPP+) .METHODS:The rat model of Parkinson disease ( PD) was established by intranigral microinjection of MPP +.The degree of microglial activation was measured by immuno-fluorescent density of OX-42 ( a microglia marker ) in the substantia nigra ( SN) .The number of viable dopaminergic neurons was determined by counting the tyrosine hydroxylase ( TH) positive neurons in the SN .The behavioral performances were re-vealed with the number of apomorphine-induced rotations , score of forelimb akinesia and vibrissae-elicited forelimb placing a-symmetry.RESULTS:Pretreatment with CoQ10 or intracerebroventricular (icv) posttreatment with minocycline alone pro-vided partial attenuation against MPP +-induced locomotor defects .Integrative therapy provided enhanced beneficial effects , and resulted in a significant attenuation of locomotor disability than any single therapy (all P<0.01).The results of immu-nohistological analysis showed that the TH positive neurons were maximally protected by integrative therapy compared with minocycline group and CoQ 10 group (P<0.01) .CONCLUSION:The integrative therapy of CoQ 10 combined with minocy-cline may offer additional therapeutic benefit to MPP +-induced hemiparkinson rat model .Such neuroprotective strategy of tar-geting different aspect of the neurodegenerative phenotypes may highlight a new therapeutic strategy for future management of PD.

Objective To investigate the clinical outcomes of the medial displacement calcaneal osteotomy with reconstruction of posterior tibial tendon insertion on navicular for the flatfoot related with accessory navicular.Methods From March 2009 to October 2011,13 patients (16 feet) with flatfoot related with accessory navicular received treatment by the medial displacement calcaneal osteotomy with reconstruction of posterior tibial tendon insertion on navicular.There were 4 males and 9 females,with an average age of 41.3 years (range,18-64 years).All patients had obvious valgus calcaneus,the angle of which was 11.3°± 1.4°.According to AOFAS ankle-hindfoot scores,the arch height,calcaneus inclination angle (CI),talocalcaneal angle (TC),talar first metatarsal angle (TMT) on the lateral weight-bearing radiograph of foot,and the talocalcaneal angle (TC),talar first metatarsal angle (TMT) on the AP view of the weight-bearing radiograph of foot,and the heel valgus alignment on axial radiographs of the hindfoot were measured on the X-ray film.Results All patients were followed up for 12 to 31 months,with the average of 16.8 months.Eleven patients (13 feet) felt no pain 6 months after operation,while 2(3 feet) felt pain after long walking.There was no complication,including infection,nerve injury,un-union,and so on.The average AOFAS ankle-hindfoot score improved from 56.4-±6.4 preoperatively to 88.1±2.8 at the last follow-up.Radio graphically,all parameters were statistically significant between pre-operation and the last follow-up,including the arch height,CI,TC,TMT modifying from 3.8±0.3 mm,9.5°±1.1°,47.3°±2.5°,17.6°±1.6° to 12.0±1.1 mm,20.1°±1.5°,32.3°±2.5°,6.8°±1.0° respectively on the lateral weight-bearing view; TC improving from 39.5°±2.3° to 26.2°±2.0°and TMT improving from 15.2°±1.7° to 6.3°±1.0° on the AP weight-bearing view.Conclusion The medial displacement calcaneal osteotomy with reconstruction of posterior tibial tendon insertion on navicular is a good choice for the treatment of flatfoot related with accessory navicular with excellent clinical outcomes.

Objective This review was aimed to provide reference for production, management and use of laboratory animals by analyzing the test results on intestinal parasitic infections of mice and rats in different provinces from 1989 to 2013 in China.The results showed that the infection rates in clean and SPF mice and rats were reduced to 10%, being better than that in the past years, but the situation was still not optimistic for the control of flagellate parasites infections.

Objective To investigate the neuroprotective effect of Dengzhan Shengmai capsule (DZSM) in rat cerebral ischemia-reperfusion injury and to explore the mechanism. Methods Rats were divided into Sham group, MCAO group, DZSM group, carbenoxolone (CBX) group and DZSM + CBX group. Each group was assessed for neurological function , infarct volume and the expression of Caspase-3 48 h after reperfusion. Connexin 43 (Cx43) expression of MCAO group was detected 3, 12, 24, 48 h after reperfusion. Results There were lower neurological deficit scores , infarct volume and the expression of Caspase-3 in DZSM , CBX and DZSM + CBX group 48 h after reperfusion when compared with those in MCAO group (P < 0.05) but Cx43 expression level in each group increased after reperfusion at each time point (P < 0.05). Expression of Cx43 was lower in DZSM, CBX and DZSM + CBX group than that in MCAO group (P < 0.05). Lower expression of Cx43 was also seen in CBX and DZSM + CBX group when compared with that in DZSM group (P < 0.05). Conclusion DZSM capsule can improve neurological function , reduce infarct volume and inhibit the expression of Caspase-3. The mechanism may be related to its inhibition of Cx43 expression.

[ ABSTRACT] AIM:To investigate the effect of triptolide on the inhibition of microglial activation in 1-methyl-4-phenyl pyridinium ( MPP+)-induced hemiparkinson disease rats.METHODS:The rat model of Parkinson disease was es-tablished by intranigral injection of MPP +.The rats were randomly divided into sham group, MPP+group, triptolide group and vehicle group.The survival of dopaminergic neurons was detected by the immunofluorescence of tyrosine hydroxylase ( TH) in the substantia nigra ( SN) .The activation of microglia was determined by immunofluorescence of OX-42 ( micro-glia marker) in the SN.The expression of chemokine receptor CX3CR1 in SN was measured by Western blotting.RE-SULTS:Intranigral injection of MPP+increased the fluorescence intensity of the microglial marker, and promoted DA neu-ron degenerative death.Immunohistological analysis showed that the OX-42 density was decreased (P<0.01) and tyrosine hydroxylase (TH) positive neurons were increased in the triptolide group (P<0.01).The expression of CX3CR1 was lower in triptolide group than that in model group (P<0.05).CONCLUSION:Triptolide may improve PA neurons func-tion in MPP+-induced rats through inhibiting CX3CR1 expression and microglial activation.

Objective To establish the a staging system based on scoring of ketamine associated urinary dysfunction (KAUD) and evaluate its diagnosis performance and clinical value.Methods A retrospective analysis of 135 admitted KAUD patients from September 2009 to September 2015 was performed.All patients were classified into three stages according to bladder and upper urinary tract impairment, and treated with subsequent strategy.The behavioral modification and pharmacotherapy was performed in stage Ⅰ which image investigation was normal.Hydrodistension and intravesical perfusion was used if pharmacotherapy failed in stage Ⅱ patients who had bladder impairment in image or urodynamics.Augmentation cystoplasty or cystectomy with urinary diversion was optional if aforementioned treatment failed in stage Ⅲ who had upper urinary tract involvement and abnormal renal function.The voided volume, micturition interval, and nocturnal void frequency as recorded in their 3-day bladder diary, PUF, ICPI and ICSI score after treatment in one month, were compared with their baseline data prior to treatment.Ketamine abusing history, renal and liver function, bladder and upper urinary tract changes, urodynamics and radioisotope renography correlation with stages were analyzed.A scoring system was established and the concordance between the clinical staging and model staging of each patient was compared.Receiver operating characteristic curve was employed to testify the sensitivity and specificity.Results There were 39, 80 and 16 patients included in each stage respectively.After subsequent strategy of treatment, voided volume, micturition interval, and nocturnal void frequency, PUF, ICPI and ICSI were improved significantly.Age (P =0.018), ketamine abusing history (P =0.006), eGFR (P =0.001), ALT (P =0.037), γ-GT (P ＜ 0.001), bladder (P =0.013) and ureteral changes (P ＜ 0.001), hydronephrosis (P ＜ 0.001) and radioisotope renography abnormality (P =0.01) were correlated with clinical stages.In total, 107 cases' stages based on scoring were accordant with clinical stages (30, 65 and 12 cases in each stage respectively).AUC of the scoring model ROC was 0.922 on cut point between Ⅰ and Ⅱ stages and AUC of the scoring model ROC was 0.978 on cut point between Ⅱ and Ⅲ stages.Conclusions We presented a scoring staging system of KAUD based on ketamine abusing history, laboratory test results, and imaging findings.It might be helpful to evaluate the progression on bladder and upper urinary tract, and provide a reference for treatment.This staging system based on scoring still needs further validation and improvement.

Objective To identify the risk factors of chronic ketamine-induced upper urinary tract dysfunction (KUUTD).Methods Medical records of 103 patients with ketamine-associated urinary dysfunction(KAUD) from Sep.2009 to Jul.2015 were retrospectively reviewed.All the patients were divided into the upper urinary tract dysfunction group(n =39)or the normal group(n =64)according to the diagnostic criteria.Indexes such as general features (age,gender,BMI),drug-abusing characteristics (abusing time,frequency,dose,abstinent period),duration of LUTS,pelvic pain and urgency/frequency symptom score(PUF score),serum parameters (WBC,ALT,AST,γ-GT),bladder diary (daytime inter-void interval,nocturia,average urine volume),urodynamic data (maximum cystometric capacity,post-void residual,maximum urethral closure pressure,maximum and average flow rate,instable contraction,bladder compliance) and imaging manifestations were compared between the two groups.Logistic regression analysis was performed for the evaluation of variables with statistical significance.Results There was significantly difference between the two groups in BMI (t =2.127,P =0.043),drug-abusing frequency (t =2.335,P =0.029),abstinent period (t =3.451,P =0.003),duration of LUTS (t =2.289,P =0.036),PUF score (t =3.030,P =0.004),γ-GT value(t =2.697,P =0.012),MCC(t =5.056,P ＜ 0.001),MUCP(t =3.015,P =0.008),BC (t =8.562,P ＜ 0.001) and imaging findings of bladder contracture (x2 =15.155,P ＜0.001).The multivariate Logistic regression analysis showed that drug-abusing frequency(OR =1.279,P =0.038),abstinent period(OR =0.394,P =0.028),γ-GT value (OR =1.007,P =0.032),MCC (OR =0.775,P =0.011)and BC (OR =0.013,P =0.008)were the key factors of KUUTD.Conclusion Consecutive and frequent drug abuse,combination with chronic obstructive cholangitis,reduced capacity and low compliance bladder are the risk factors of upper urinary tract dysfunction associated with ketamine abuse.

Objective Plasma circulating DNA can be em-ployed in place of bone marrow examination for the auxiliary diagnosis of leukemia.This study aimed to explore the clinical application of the plasma DNA level in evaluating the effect of chemotherapy on chronic leukemia. Methods We collected blood samples from 52 patients with chronic myelogenous leukemia (CML) (33 in the chronic phase, 7 in the acceleration phase, and 12 in the blast phase) , 85 with chron-ic lymphocytic leukemia (CLL) (28 with complete remission, 27 with partial remission, and 30 with no remission), 4 patients with hairy cell leukemia (HCL), and 80 healthy subjects.We simultaneously obtained plasma DNA and recombinant plasmid DNA using the BI-LATEST DNA Kit and examined the human β-actin gene and the level of plasmid DNA by real-time quantitative PCR. Results Before chemotherapy, the median value of plasma DNA was 149.46(30.63-496.91)ng/ml in the CML and 101.54(69.10-258.14) ng/ml in the CLL patients, both significantly higher than in the healthy controls (19.05[12.67-25.92]ng/ml) (P<0.01).After chemotherapy, the plasma DNA level of the CML patients was remarkably decreased, but still higher than that of the controls ( P<0.01).The CML patients in the chronic phase showed a markedly higher level of plasma DNA (302.89[93.33-541.52]ng/ml) than those in the blast phase (43.19[23.54-70.03]ng/ml) and acceleration phase (28.11[16.21-92.07]ng/ml) (P<0.05).The CLL patients with CR exhibited a significantly lower level of plasma DNA (24.29[14.64-30.74]ng/ml) than those with PR (106.88 [96.23-143.25]ng/ml) and NR (460.73[284.57-653.38〗ng/ml) (P<0.01), but all dramatically higher than that of the healthy controls (P<0.01) Conclusion The quantification of plasma DNA has a clinical application value in evaluating the effect of chemo-therapy on chronic leukemia.

Skin sensitization (allergic contact dermatitis, ACD), is a serious condition caused by small reactive molecules and is characterized by a delayed-type hypersensitivity .Animal tests were usually used in the evaluation of sensitizing potential of chemical substances in the past .However , there is an increasing interest from the public for reducing and ultimately replacing animal tests .The European Union (EU) has posed a ban on animal testing of cosmetic ingredients that includes skin sensitization since 2013.Therefore, alternative in vitro tests are the main tendency in chemical substances and cosmetic sensitizing potential research in the future .In this study, different kinds of in vitro test methods that were adopted by OECD or on research (LLNA, DPRA, KeratinoSens TM, h-CLAT) were reviewed through recent years literature , comprehensive introduction and evaluation were made to obtain reliable hazard and potency information on potential skin sensitizers .

Clustered regularly interspaced short palindromic repeats (CRISPR) is an acquired immune system existing in archaea and bacteria with the long-term process of evolutionary.CRISPR/Cas9 gene editing system is a new type of gene editing technology developed based on the system.CRISPR/Cas9 is a more efficient method for gene targeting than the previous methods.It has been successfully applied for gene-modified of eukaryotes since 2012,but the reports about pathogenic microorgaisms are rarely.Here,the research progress in the structure,mechanism of CRISPR/Cas9 system and its applications on pathogenic microorgaisms is reviewed.