Objective To investigate expression of 14-3-3 sigma gene in patients with breast cancer and its clinical significance.Methods Expression of 14-3-3 sigma gene was semi-quantitated by using RT-PCR and western-blot in 40 specimens of breast cancer and 18 specimens benign breast disease tissue.Results Of 40 cases of breast cancer 35(87.5%)were negative for 14-3-3 sigma gene in RT-PCR,32(80%)were negative in Western-blot,and 31(77.5%)were negative in both RT-PCR and western-blot.Besides,the expression in 2 cases was down-regulation in both the 2 method.In 18 specimens with benign breast disease tissue the expression of 14-3-3 sigma gene was detectable,which was demonstrated by RT-PCR or western-blot.Conclusion Inactivation and down-regulation of 14-3-3 sigma gene is a frequent event in breast cancer,and it may contribute to diagnosis of breast cancer.

105 copies/mL would develop liver-related complications, such as cirrhosis, hepatocellular carcinoma and liver failure. Available evidences indicate that control of HBV replication with antiviral drugs can decrease the incidence of these complications and mortality.

Objective To investigate the effects of intrathecal (IT) neostigmine on the excitatory amino-acid content in the L4-5 segment of spinal cord in a rat model of incisional pain. Methods Thirty-two male SD rats weighing 250-270 g were anesthetized with intraperitoneal pentobarbital 40 mg ? kg-1 . Intrathecal catheter was placed with the tip reaching the lumbar region according to the method of Yaksh. Five days later an 1 cm long incision was made in the plantar region of the right hind paw under isoflurane anesthesia according to the method of Brennan. Pain behavior was assessed at 1h after incision by cumulative pain score. The animals were randomly divided into four groups with 8 animals in each group: Ⅰsham operation group received IT artificial cerebro-spinal fluid (ACSF) 20 ?l but no incision was made in the hind paw; Ⅱ control group received ACSF 20 ?1 30 min before incision was made; Ⅲ postoperative neostigmine group received IT neostigmine 10 ?g 30 min after incision; Ⅳ preoperative neostigmine group received IT neostigmine 10 ?g before incision. 2h after incision the animals were decapitated and lumbar segment of spinal cord was removed for determination of aspartate and glutamate contents by high performance liquid chromatography (HPLC) with fluorescence detection. Results The cumulative pain scores in group Ⅱ were significantly higher than those in group Ⅰ (P 0.05) . Conclusion The decline in the increased excitatory amino-acid contents in spinal cord induced by incisional pain is involved in the mechanisms of analgesia provided by IT neostigmine.

Objective To explore the possibility of heterogenous gene to express in juvenile Schistosoma japonicum and the application of electroporation in transformation of schistosomulae. Methods The plasmids of pEGFP-C1 were introduced into mechanically transformed schsitosomula with electroporation. The presence, transcription and translation of the transgene in electroporated schistosomula were confirmed by PCR, RT-PCR and Western blotting analysis respectively using the genomic DNA, total RNA and protein extracted and isolated from schistosomula cultured in vitro for 48 hours. Meanwhile, localization of EGFP within electroporated schistosomula was performed with confocal laser scanning microscope. Results 760 bp and 276 bp amplified products by PCR and RT-PCR were found coincident with the expected size and expression of EGFP gene in elctroporated schistosomula was confirmed by Western blotting. Fluorescence of EGFP was localized in tegument and subtegument of the electroporated schistosomula with confocal microscopy, especially in the anterior part of the worm. Conclusion The heterogenous gene of EGFP has been successfully introduced into juvenile S. japonicum by electroporation and the expression of transgene was confirmed with molecular and microscopical methods.

Objective To investigate correlation between the results of drug susceptibility and the extent of drug-resistances in Mycobacterium tuberculosis clinical isolates. Methods Liquid culture and MTT test were used. Twelve anti-TB drug MICs and drug susceptibility testing of the 163 MTB strains from random clinical isolates were detected, which including RFP, INH, SM, EBM, OFLX, LVFX, MOX, AMK,CPM, PTA, CLA and PAIN. Results There are 67% (42/62) Mycobacterium tuberculosis strains resistant to SM, 63% (51/81) Mycobacterium tuberculosis strains resistant to INH, 77% (50/65) Mycobacterium tuberculosis strains resistant to RFP, 41% ( 15/37 ) Mycobacterium tuberculosis strains resistant to AMK,41% (12/29) Mycobacterium tuberculosis strains resistant to CPM, 20% (12/60) Mycobacterium tuberculosis strains resistant to EMB and 43% (25/58) Mycobacterium tuberculosis strains resistant to OFLX which MICs were equal to or more than 16 μg/ml, 8 μg/ml, 8 μg/ml, 16 μg/ml and 4 μg/ml, 4 μg/ml and 8 μg/ml,respectively. There were significant differences in the MICs of OFLX, LVFX and MOX in OFLX resistant strains (2-128, 1-32 and 0.0625-1 μg/ml, respectively) by ANOVA ( F = 16.874, P ＜ 0.001 ). The MICs of SM, INH, RFP, EMB, OFLX, AMK and CPM in isolates resistant to six or seven drugs (0.5-128,2-64,0.25-128,1-32,1-64,0.5-128 and 1-128 μg/ml,respectively) were higher than those (0.25-128,0.0625-64,0.25-32,0.25-2,0.125-2,0.5-4 and 1-4 μg/ml,respectively) in isolates resistant to one or two drugs (F=20.066, 40.499, 47. 197, 70.373, 91.432, 41.840 and 21.547, respectively, P ＜0.05). The MICs of SM, INH, RFP and EMB in isolates resistant to four drugs (1-128,2-64,0.25-128 and 1-32 μg/ml,respectively ) were higher than those ( 0.25-128,0.0625-64, 0.25-64 and 0.25-2 μg/ml,respectively) in isolates resistant to one or two drugs (F = 26.242, 23.563, 31.541 and 64.469,respectively, P ＜0.05).The MICs of RFP in MDR isolates (2-64 μg/ml) were higher than those (0. 25 μg/ml) in other resistant isolate except M DR isolates (F = 5.613, P <0.05). Conclusions The study shows that there are associations between the results of routine drug susceptibility testing and the resistant extent of anti-TB drugs. This could help doctors select more effective anti-TB regimen for TB patients according to the correlations.

Objective To establish a method of judge the pyrazinamide(PZA) susceptibility for Mycobacterium tuberculosis(MTB) with 24-hole micro-liquid culture silica gel color plate,and evaluate the clinical application value of this method.Methods According to the result of MGIT960,to detect PZA drug susceptibility for 30 MTB clinical isolates of whose PZA susceptibility were known with silica gel color plate.The effects of different pH value,different inoculation concentrations on the results were observed,and the optimum detection conditions were discussed.Finally,the 98 MTB clinical isolated of whose PZA susceptibility were unknown were simultaneously detected by gel color plate and MGIT960,the sensitivity,specificity and accuracy were judged by PZA drug sensitivity.Results The best pH was 5.8-5.9 and the best concentration was 2.5×10-1 mg/mL in gel color plate.The best results were read after 7-14 d.The sensitivity,specificity and accuracy were 95.50%,96.30%,and 98.21% respectively just at PZA critical concentration was 100 μg/mg.The sensitivity,specificity and accuracy were 90.90%,92.59%,and 91.84% respectively just at PZA critical concentration was 200 μg/mg.Conclusion The PZA susceptibility for MTB with 24-hole micro-liquid culture silica gel color plate is accurate,rapid,low-cost and simple.

Objective To discuss the clinical application value of microplate allochroic silica gel assay for rapidly detecting rifampicin(RIF)-resistance of Mycobacterium tuberculosis(MTB).Methods Fifty MTB clinical isolates preserved in the tuberculosis(TB) laboratory of Nantong Municipal Sixth People's Hospital were detected RIF-resistance by using the microplate allochroic silica gel assay and compared with the Bactec MGIT960 method.Then,the RIF susceptibility test in 40 clinical sputum smear-positive specimens were simultaneously detected by using the microplate allochroic silica gel assay and Bactec MGIT960.Finally the sensitivity,specificity and accuracy of the test results were compared.Results The optimal inoculation volume of MTB was 10-3 mg/mL,the optimal detection time was 7-10 d and the judging critical value of the RIF minimum inhibitory concentration(MIC) Was 1.00 μg/mL by microplate allochroic silica gel assay.With the Bactec MGIT960 test as the gold standard,the sensitivity,specificity and accuracies of microplate allochroic silica gel assay for RIF-resistance susceptibility test of smear-positive specimens were 94.12％,100％ and 97.37％ respectively.Conclusion Microplate Allochroic silica gel assay can be used for directly detecting the MTB sensitivity to RIF of in sputum specimens.

Objective: To study the distribution and related factors of curative care expenditure (CCE) of injury in Gansu Province in 2017. Methods: Based on the "A System of Health Accounts 2011 (SHA 2011)", the curative care expenditure of injury in Gansu Province was calculated and analyzed. The five-stage stratified cluster sampling method was adopted to extract 149 medical and health institutions, 120 township hospitals (including community health service centers), 150 individual clinics and 600 village clinics (including community health service stations). The top-down allocation method was used to calculate the cost of injury treatment in Gansu Province, and the influencing factors were analyzed by multiple linear regression. Results: In 2017, the CCE of injury in Gansu province was 3.831 billion yuan, and the expense in general hospitals was 2.708 billion yuan. Among them, the cost of lower limb injury and head injury were 1.090 and 0.847 billion yuan. People aged 40 to 69 years old spent 1.901 billion yuan on injury treatment, and the CCE of injury treatment for men and women were 2.422 and 1.409 billion yuan respectively. The results of multiple linear regression showed that hospitalization expenditure was significantly associated with length of stay, operation, hospital grade, age, payment method and gender (P<0.001). Conclusion The economic burden of injury in Gansu Province is relatively heavy, so it is necessary to focus on preventions for different groups and costly injury sites.

ObjectiveTo establish an allele-specific polymerase chain reaction （PCR） method for identifying Scolopendra dispensing granules， so as to ensure the quality and therapeutic effects of Scolopendra and its preparations. MethodThe primer interval suitable for the PCR was selected based on the cytochrome c oxidase subunit 3（COX-3） gene sequence of Scolopendra， and the single nucleotide polymorphism （SNP） loci of Scolopendra and its adulterants were mined from the interval for the design of specific primers. The samples of Scolopendra and its adulterants were collected. The PCR system was established and optimized regarding the annealing temperature， cycles， Taq enzymes， DNA template amount， PCR instruments， and primer concentrations， and the specificity and applicability of this method were evaluated. ResultThe PCR system was composed of 12.5 μL 2×M5 PCR Mix， 0.4 μL forward primer （10 μmol·L^-1）， 0.4 μL reverse primer （10 μmol·L^-1）， 2.5 μL DNA template， and 9.2 μL sterile double distilled water. PCR parameters： Pre-denaturation at 94 ℃ for 3 min， 30 cycles （94 ℃ for 20 s， 62 ℃ for 20 s， 72 ℃ for 45 s）， and extension at 72 ℃ for 5 min. After PCR amplification with the system and parameters above， the electrophoresis revealed a bright band at about 135 bp for Scolopendra and no band for the adulterants. ConclusionThe established allele-specific PCR method can accurately identify the medicinal materials， decoction pieces， and standard decoction freeze-dried powder of Scolopendra， as well as the intermediates and final products of Scolopendra dispensing granules， which is of great significance for ensuring the quality and clinical efficacy of Scolopendra and its preparations.