Objective: To study the taste of unprocessed and processed Siegesbeckiae puescens (SP) and establish a taste discrimination model. Methods: The sour, bitter, astringent, salty, and sweet values of 12 batches of unprocessed and processed SP were measured by electronic tongue using a paired t-test, principal component analysis (PCA), and the linear discriminant factor analysis (LDA) to study the changes of the taste of unprocessed and processed SP. Results: The paired t-test showed that the bitterness and saltiness of processed SP decreased; Astringency, sweetness, and sourness did not significantly change. The PCA can distinguish unprocessed and processed SP. The model of the taste of unprocessed and processed SP was established through the LDA, and conduct cross validation; Its correct rate reached 100%. The fingerprint of taste of unprocessed and processed SP was established through radar-graph. Conclusion: Electronic tongue can identify sour, bitter, astringent, salty, and sweet values of traditional Chinese medicine accurately. It can reflect the numerical change of taste of unprocessed and processed SP combining with statistical method. Based on it, it can be inferred that the relationship between the change of taste and clinical efficacy before and after processing of SP. Besides, the discriminant model can be used to distinguish unprocessed and processed SP.

As the major commercial source of natural rubber,Hevea brasiliensis attracts much attention.However,the heterozygous nature,long breeding cycle are strong limitations for conventional breeding.While genetic engineering,which can be used to widen the germplasm base and produce desirable agronomic traits quickly and efficiently,offers a viable alternative approach to complement traditional breeding.Comprehensive analysis indicates that in the past two decades,with calli derived from immature anther or integumental tissues of immature fruit as receptors,both biolistic and Agrobacterium-mediated transformation methods were employed for developping rubber genotypes with improved latex yield,tolerance to tapping panel dryness syndrome,producing high-value recombinant proteins,etc.Being recalcitrant to tissue culture,the transformation efficiency of Hevea is comparatively low,and the procedures are still needed to optimize.Finally,breeding objectives and strategies to improve transformation efficiency were also proposed in the review.

Objective To develop a BP26 recombinant BCG (rBCG-BP26) vaccine,and to observe the effects of rBCG-BP26 on CD4+,CD8+ T cells in immunized mice.Methods The recombinant shuttle vector pMV261-Ag85B-BP26 was constructed by using traditional molecular biological technology.The recombinant strains were obtained by kanamycin resistance screening and PCR identification after electroporation.Western blotting was used to detect the expression of recombinant BP26 vaccine in immunized mice.Safety experiment was carried out in three different groups:the target experiment(rBCG-BP26) group,the positive control(BCG) group and the negative control(PBS) group,15 BALB/c mice in each group.Intradermal inoculations of 100 μl rBCG-BP26 [containing 106 colony forming units(CFU)],BCG,and PBS were carried out,respectively.Signs of mice in each group were observed.After immunization for 10,20,30,and 40 days,body weight was weighed,and tail blood was collected to observe the change of peripheral blood CD4+ and CD8+ T cells by flow cytometry.Results The rBCG-BP26 was successfully constructed.The expression of BP26 protein was detected in the liquid medium and the bacteria cells.The results of safety test analysis showed that there were no significant differences in signs and body weights(F=2.468,0.331,1.520,0.739,all P＞ 0.05),between PBS group[ (19.24 ± 0.54),(21.37 ± 0.66),(22.83 ± 0.62),(25.06 ± 0.37)g],BCG group[ (19.90 ± 0.02),(21.53 ± 1.57),(21.95 ± 0.55),(24.70 ± 0.39)g]and rBCG-BP26 group[ (19.16 ± 0.55 ),(20.89 ± 0.20),(22.15 ± 0.76),(24.60 ± 0.64)g].The results of flow cytometry showed that the percentages of CD4+ T cell level were lower in BCG group(26.70％,33.07％) and rBCG-BP26 group( 13.40％,26.70％) than that of the PBS group(33.85％,29.33％) and the values of CD4+/CD8+ T cells increased in rBCG-BP26 group (0.69％,1.27％,1.57％,1.70％ ) 10,20 and 30 days after immunization.Conclusions Recombinant BCG-BP26 vaccine strain can express brucella BP26 protein efficiently.Furthermore,its virulence is mild,and it can activate CD4+,CD8+ T cells in the body.It can be used as one of candidate vaccine strain against brucellosis.

ObjectiveTo investigate the concentration of water fluoride and the application of water defluoridation projects in Suizhou,and to provide a basis for water improvement and prevention of the disease.MethodsCross-sectional study was carried out to investigate completely the water defluoridation projects built between 1986 and 2009 in the area under Suizhou's jurisdiction; 3 source water and terminal water samples of the projects in use were collected,respectively; 1 water sample was collected,respectively in the 3 projects with out-ofcommission or discarded water sources that used to be major water supply,for detection of fluoride,chloride,total hardness,heavy metals and other indicators.ResultsA total of 21 projects were investigated,14 projects were in normal use (66.7％),7 projects were out of order and abandoned (33.3％).Of the 14 projects normally used,13projects with water fluoride concentration ≤ 1.0 mg/L(92.9％)and 1 project with water fluoride concentration higher than 1.0 mg/L(7.1％),the concentration of water fluoride in the 7 abandoned projects was higher than 1.0 mg/L(100％).Arsenic and lead levels of all the water samples were normal.There were two water samples with iron content exceeded the standard,and one total hardness of water samples exceeded the standard in the normally used 14 peripheral water sources.One chloride content exceeded the standard,one manganese content exceeded the standard,two iron content exceeded the standard and three total hardness exceeded the standard in the seven abandoned projects.ConclusionsSome defluoridation projects are stopped using and abandoned in Suizhou,and the water fluoride exceeds the standard.

Objective To construct the bp26 deletion mutant of Brucella vaccine strain M5-90 (M5-90Δbp26),to compare its immunogenicity with parental strain(M5-90),and to develope a new candidate vaccine strain of Brucella melitensis with reduced virulence that can be used to distinguish vaccinated livestock from infected animals.Methods Mutant vaccine strain of Brucella melitensis was constructed by conventional molecular biology techniques then the genetic stability of mutant M5-90Δbp26 was tested and its conventional bacteriological nature was identified; 1.0 × 109 CFU/2 ml doses of M5-90Δbp26 strain and the parental strain were used to vaccinate 3 sheep; sera were analyzed for reactivity against BP26 by Western blotting and for agglutination activity; to analyze the virulence of mutant and parental strain,mice were injected with 1.0 × 106,6.0 × 106 and 2.0 × 107 CFU/0.2 ml doses of M5-90 and M5-90Δbp26,respectively,and clinical symptoms were monitored and the death of mice was recorded.Results The M5-90Δbp26 was successfully generated and reversion was not observed in 15 generations.The size of PCR products was 629 bp while the parental strain was 1279 bp.The sequence analysis showed a 650 bp missing in M5-90Δbp26.The conventional bacteria identification tests confirmed that the mutant was depth variant strain,including mono-specific antiserum M type transformed into R,and the BK2 phage based splitting assay converted from the positive to the negative.Western blotting showed the purified BP26 protein was recognized by the serum against the parental strain while not by the serum against M5-90Δbp26 strain.Agglutination test showed the level of the serum antibody induced by M5-90Δbp26 strain(1:50) was significantly lower than that of serum induced by parental strain(＞ 1:800).Virulence test showed that M5-90Δbp26 strain was less virulent than parental strain.Conclusions M5-90Δbp26 has been successfully constructed.M5-90Δbp26 of Brucella melitensis has the characteristic of reduced virulence and has a potential as brucellosis candidate vaccine strain permitting serological discrimination between diseased and vaccinated livestock.

Objective To determine the difference of macrophage RAW264.7 apoptosis induced by Brucella melitensis virulent strain 16M and attenuated strain M5-90 and elucidate the regulatory role of caspases 3,8 and 9.Methods The best multiplicity of infection (MOI) was determined through kinetic analysis of Brucella melitensis strain 16M and M5-90 induced mouse macrophages apoptosis(bacterium ∶ cell =100 ∶ 1,50 ∶ 1,10 ∶1).The infection model was established using the best MOI =50 ∶ 1.The numbers of in vivo bacteria by colony formation units were calculated after macrophages were infected for different times,including 2,4,8,12,24 and 48 h,and the infected cells were collected.The ratios of apoptosis were detected and the regulation of caspases 3,8 and 9 in apoptosis pathway was elucidated by flow cytometry.Results The numbers of 16M in vivo bacteria were 105.4,104.8,105.8,106.5,108.0 and 109.0,respectively and of M5-90 were 106.1,106.2,106.4,106.3,106.1 and 105.0,respectively.The number of in vivo bacteria of 16M was significantly increased than that of M5-90 after infected for 24 h to 48 h.The ratios of apoptosis induced by 16M after infected for 2,4,8,12,24 and 48 h was (2.67 ± 0.09)％,(13.13 ± 0.30)％,(6.56 ± 0.42)％,(6.49 ± 0.28)％,(16.07 ± 0.86)％ and (24.23 ± 1.67)％,respectively,and by M5-90 was (3.62 ± 0.02)％,(32.01 ± 2.59)％,(17.58 ± 0.44)％,(16.09 ± 0.10)％,(62.53 ± 2.70)％ and (85.53 ± 0.15)％,respectively,and by control group was [(1.90 ± 0.20)％,(1.92 ±0.16)％,(1.99 ± 0.03)％,(2.48 ± 0.11)％,(3.56 ± 0.07)％,(5.26 ± 0.33)％].The differences were statistically between groups in same time.The Brucella melitensis vaccine strain M5-90 was more powerful than virulent strain 16M in respect of inducing macrophage apoptosis after infected for 24 to 48 h.Twenty-four hours after infection,the expression of caspases 3,8 and 9 was (1.47 ± 0.05)％,(1.52 ± 0.02)％ and (2.47 ± 0.12)％,respectively,in control group and the expression was (9.70 ± 0.46)％,(6.08 ± 0.56)％ and (35.08 ± 1.64)％,respectively,after infected for 24 h induced by M5-90.The expression of caspases 3,8 and 9 was significantly higher than that control group (P ＜ 0.01).Twenty-four hours after given caspases 3,8 and 9 inhibitor,apoptosis rate in control group was (66.72 ± 1.28)％,in M5-90 group was (22.58 ± 0.55)％,(53.15 ± 1.85)％ and (29.18 ± 0.23)％,respectively,and compared with control group,apoptosis rate of caspases 3,8 and 9 was significantly lower(P ＜ 0.01).Conclusions Apoptosis of macrophage can be induced by Brucella melitensis virulent vaccine strain 16M and attenuated strain M5-90.M5-90 is stronger than that of strain 16M.Caspases 3,8 and 9 can regulate macrophage apoptosis after M5-90 infection.

Objective To investigate the clinical value of ~(99)Tc~m-ethylenedicysteine (EC) diuretic renography (DR) in pre-and post-operative pediatric congenital hydronephrosis.Methods The DR with injection of Furosemide at 15 min of forty children with hydronephrosis was retrospectively studied.The preoperative renal blood perfusion rate (BPR),effective renal plasma flow (ERPF),grade of hydronephrosis,renogram and renal dynamic imaging of pre- and post-operative kidneys were compared.The t-test and Mann-Whitney test were used for data analysis.Results (1) Of 40 pathological kidneys,the BPR increased 5.99% (t=-5.13,P＜0.01)from pre-operative to post-operative:(34.05±11.07)% to (40.04±8.56)%.The ERPF increased 12.48 ml/min(t=-4.35,P＜0.01) from pre-operative to post-operative:(57.81±34.32)ml/min to(70.29±5.37)ml/min.(2)The grade of hydronephrosis of 40 pathological kidneys improved significantly(Z=-2.64,P＜0.01) with the mean sum of ranks of 47.21 pre-operatively to 33.79 post-operatively.(3) As the hydronephrosis worsened,the collecting system became bigger,the renal parenchyma became thinner,the extent of intrarenal parenchymal photopenia became larger and the response to diuretic challenge in pathological kidneys decreased or became totally irresponsive.(4)Thirty-seven cases of obstruction at ureteropelvic junction (UPJO) and 3 cases at ureterovesical junction (UVJO) were diagnosed by DR,which were all confirmed by surgery.Conclusions DR is a reliable method to evaluate pediatric congenital hydronephrosis.It can accurately reflect the grade and (or) severity of the disease,guide therapy and assess the therapeutic success of operation.

Objective Prospective comparison was done on concurrent chemo-radiotherapy and se- quential chemo-radiotherapy for unresectable stageⅢnon-small cell lung cancer(NSCLC) and to evaluate three different regimens of concurrent chemo-radiotherapy.Methods Ninety-six such patients were ran- domized into four groups:1.sequential chemo-radiotherapy group received two cycles of induction chemother- apy with 40 mg/m~2 of cisplatin on D 1-3,29-31 and 100 mg/m~2 of etoposide on D 1-3,29-31 before conven- tional radiotherapy,2.concurrent chemo-radiotherapy group 1 received 100 mg/m~2 etoposide on D 1-3 and DDP 40 mg/m~2 on D 1-3,D 29-31,iv.drip,3.concurrent chemo-radiotherapy group 2 received concurrent chemotherapy with 40 mg/m~2 of paclitaxel every Monday during conventional radiotherapy,4.concurrent chemo-radiotherapy group 3 received concurrent chemotherapy with 40 mg/m~2 of paclitaxel every Monday during three-dimensional conformal radiotherapy.All patients were irradiated with 2.0 Gy/fraction,5 frac- tions/week,to a total dose of 60-64 Gy.They all received two cycles of consolidation themotherapy with 40 mg/m~2 of cisplatin on D 1-3 and 100 mg/m~2 of etoposide on D 1-3.Results The overa/1 response rate was 67%,71%,71% and 79% for sequential ehemo-radiotherapy group,concurrent chemo-radiotherapy group 1,2 and 3,respectively.There was a significant difference between the concurrent chemo-radiotherapy and sequential chemo-radiotherapy(P＜0.05).The 1-,3-and 5-year overall survival rate(OS) was 54%,8% and 4%;71%,17% and 8%;79%,17% and 8%;83%,46% and 13%,respectively for the four groups. The difference among all these groups(P=0.017) was significant.It was also significant between the con- current chemo-radiotherapy group 1 and 3 (P=0.046).The difference of distant metastasis rate among all the groups was statistically insignificant (P＞0.05) also was the difference of toxicity (P＞0.05),but the severe toxicity of concurrent chemo-radiotherapy groups 1 and 2 were higher than the sequential chemo-radio- therapy group and concurrent chemo-radiotherapy group 3.Conclusions Better locoregional progression- free survival and overall survival of unresectable stageⅢnon-small cell lung cancer could be achieved by concurrent chemo-radiotherapy as compared with sequential chemo-radiotherapy though at the expense of in- crease in toxicity.With the combination of concurrent chemo-radiotherapy and conforrnal radiotherapy,the o- verall survival rate could be much improved with miider toxicity.

AIM: To investigate therapeutic efficiency of amniotic extraction on dry eye in rabbit model induced by topical benzalkonium chloride (BAC).
METHODS: Totally 26 rabbits (26 right eyes) with dry eye model were studied and divided into two groups:group A (control group with PBS eye drops, n = 13) and group B ( amniotic extraction group, n = 13). Another two rabbits were chosen as normal control. The SchirmerⅠ tests ( S Ⅰ t) and corneal fluorescein staining ( FL) were made, and the tear total protein content, amylase activity, lactoferrin, lysozyme contents, goblet cell density were performed in two groups before treatment and 1, 2, 4 and 8 wk after treatment.
RESULTS: There were significant differences in SIT, FL scores, lysozyme activity and goblet cell density among different groups at different time points (P<0. 05). But, there was no significant differences in SⅠt, FL scores, lysozyme activity and goblet cell density between two groups before treatment (P>0. 05). After 8wks' treatment with PBS, the mean differences of the group A showed great changes in SⅠt, lysozyme and goblet cell density compared with those before treatment ( P < 0. 05); but there was no significant differences in FL scores compared with those before treatment (P>0. 05). As for group B, 8wks after treatment, there were statistical changes in SⅠt, FL, lysozyme (P<0. 05); but there was no significant differences in goblet cell density compared with those before treatment ( P > 0. 05). It was evident that statistical differences were observed in S Ⅰ t, FL scores, lysozyme activity and goblet cell density between two groups at each time point (P<0. 05). However, there were no significant differences in total protein, lactoferrin, amylase activity at different time points (P>0. 05). Meanwhile there was no significant differences in total protein, lactoferrin, amylase activity between two groups before treatment ( P > 0. 05 ). But there were significant differences in total protein, lactoferrin, amylase activity between two groups after 4 and 8 wks'treatment (P<0. 05).
CONCLUSION: Amniotic extraction has significant therapeutic effect on the dry eye in rabbit model.

OBJECTIVETo study the clinical results of needle Dao for the treatment of subacromial impingement syndrome.

METHODSFrom September 2013 to August 2014, 90 patients with subacromial impingement syndrome were treated, including 61 females and 29 males, with the average age of (55.21±8.30) years old(ranged, 45 to 65 years old), and mean course of disease (3.49±2.98) months (ranged, 4 weeks to 7 months). Twenty-six patients had pains in left shoulder, and 64 patients had pains in right shoulder. Under local anesthesia, needle Dao was performed to release the pain point at the lower edge of the acromion and the subacromial adhesion. By observing the abduction of shoulder joint, anterior flexion and lifting, internal rotation and external rotation, shoulder pain score, the number of cases cured, markedly effective, effective and ineffective after treatment were counted, and the curative effect of needle Dao for the treatment of subacromial impingement syndrome was evaluated.

RESULTSAll the patients completed the treatment successfully without obvious adverse reactions. All the patients were followed up for more than 12 months. The visual analogue scale(VAS) decreased from preoperative 7.72±5.42 to postoperative 1.35±0.78(=15.89, <0.01). The Constant score at the 12th month after operation increased from preoperative 58.89±15.63 to 83.17±19.11(=28.48, <0.01). Overall efficacy evaluation was that, 46 cases were cured, 33 cases were markedly effective, 11 cases were effective, and 0 cases were invalid.

CONCLUSIONSThe advantage of needle Dao in the treatment of subacromial impingement syndrome is the minimally invasive treatment of the underlying lesions. The disadvantage is that it can not deal with the impact caused by bone abnormalities.