Objective To construct the vector that expresses the fusion protein of HIV-Tat protein and red fluorescent protein(mCherry) in mammalian cells,and observe by fluorescence microscopy the intracellular transduction and localization of recombinant protein in cells,in order to obtain a useful tool for the study of the uptake mechanism and intracellular localization of HIV-TAT.Methods With the designed primer coding mCherry sequence,the mCherry gene was amplified by PCR with the vector pmCherry-C2 as template,and inserted into vector pET14b-His-TAT to construct the expression vector pET14b-His-TAT-mCherry.The constructed vector was then transformed into E.coli BL21(DE3),which had been identified by PCR and double digested with restriction endonuclease,followed by sequencing.After IPTG induction,the recombinant protein of His-TAT-mCherry was lyzed and analyzed with SDS-PAGE.Purified His-TAT-mCherry recombinant protein was added to Hela cells and the fluorescence was observed to evaluate the transduction efficiency.Results The results of identification by PCR,digestion with restriction endonuclease and sequencing indicated that the vector His-TAT-mCherry was correctly constructed.His-TAT-mCherry fusion protein was expressed in mammalian Hela cell line and purified successfully,and the fusion protein showed cellular transduction activity.It was found by fluorescence microscopy that the red fluorescence protein located mainly over the cytoplasm,and also the membrane to some extent.Conclusion The expression vector is successfully constructed for HIV-TAT labeled with mCherry sequence.Effective expression and purification of this fusion protein is achieved.It has been observed that the constructed vector may be expressed in mammalian Hela cell under active condition.Thus,it might be useful in the study of uptake mechanism and intracellular localization of HIV-TAT.

Objective:To investigate the regulation and mechanism of chronic Trichinella spiralis ( Ts) infection on liver immunopathology in mice co-infected with Plasmodium berghei ANKA( PbA). Methods:According to body weight, 64 specific pathogen free female Kunming mice (6 - 8 weeks old, weighting 22 - 25 g) were divided into 4 groups by using random number table method. Control group: uninfected; Ts group: mice were mono-infected with 30 Ts larvae by oral gavage on day 0; PbA group: mice were mono-infected with 1 × 10 6PbA-infected red blood cells in 0.1 ml of phosphate buffer (PBS) administered by intraperitoneal injection on day 121; co-infected ( Ts+PbA) group: mice were infected with 30 Ts larvae by oral gavage and intraperitoneal injected with 1 × 10 6PbA-infected red blood cells in 0.1 ml PBS on day 121 after Ts infection. There were 16 mice in each group, in which 10 mice in each group were monitored for the survival rate. The peripheral red blood cell parasitemia of PbA group and Ts + PbA group were monitored every other day by light microscope examination of Giemsa-stained thin tail-blood smears from day 3 after PbA infection. Mice were sacrificed at day 135 after Ts infection and/or at day 15 after PbA infection, the mouse body weight and liver weight were measured, and the liver index were calculated. Ts-infected mice were monitored by a light microscope examination of diaphragm compression slide. Under a light microscope, the liver pathology and liver fibrosis of mice were observed and compared with hematoxylin-eosin (HE) staining and Sirius red staining, respectively. The F4/80 + Kupffer cells in liver of mice were examined by immunohistochemical staining. Results:After infection with Ts or PbA, Ts larvae cysts were observed in diaphragm tissues and PbA were observed in red blood cells under the light microscope. After PbA infection, there was no significant difference in survival rate between PbA group and Ts+ PbA group ( P > 0.05). Compared with PbA group, the peripheral red blood cell parasitemia was significantly decreased in Ts+ PbA group on days 11 and 15 after PbA infection (%: 27.104 ± 7.623 vs 45.032 ± 9.849, 60.218 ± 2.776 vs 76.778 ± 6.351, P < 0.05), and the liver index and the liver pathology score were significantly decreased in Ts+ PbA group ( P < 0.05). Sirius red staining showed that the positive area of liver fibrosis in Ts+ PbA group was significantly higher than that in PbA group ( P < 0.05). Immunohistochemical staining showed that the average optical density value of F4/80 + Kupffer cells in Ts+ PbA group was significantly higher than that in PbA group ( P < 0.01). Conclusion:Chronic Ts infection may reduce the peripheral red blood cell parasitemia, increase F4/80 + Kupffer cells expression in liver, and attenuate liver pathology in mice co-infected with PbA.

Objective To investigate the value of Q-analysis real-time elasticity in differentiating between benign and malignant thyroid nodules. Methods Eighty-six thyroid nodules in 62 patients with pathologic diagnosis were included in this study and were examined using Q-analysis real-time elasticity. The real-time elasticity features were observed and the quantitative index including the whole elasticity rate and the local elasticity rate were compared between benign and malignant nodules. Results There were 51 benign and 35 malignant nodules according to histopathological examination. The Q-analysis curve of real-time elasticity of benign nodules was smoother and with lower peak, compared with that of malignant nodules. The whole elasticity rate of malignant nodules were significantly higher than that of benign nodules (3.59±0.84 vs 2.32±0.56, P=0.000). And the local elasticity rate of malignant nodules were significantly higher than that of benign nodules (3.96±1.32 vs 2.39±0.58, P=0.000). The cutoff point of whole elasticity rate for the differential diagnosis was 3.25 with sensitivity, specificity and diagnostic accuracy as 71.4%, 96.1% and 86.0% respectively. The cutoff point of local elasticity rate for the differential diagnosis was 3.45 with sensitivity, specificity and diagnostic accuracy as 68.6%, 96.1% and 84.9% respectively. The diagnostic efficiency of whole elasticity rate and local elasticity rate had no significant difference (P=0.591).Conclusions Q-analysis real-time elasticity could provide the real-time elasticity features of thyroid nodules. The whole and local elasticity rate as the quantitative index contributed to the differential diagnosis of benign and malignant thyroid nodules.

OBJECTIVETo examine the effect of aqueous extracts of Scutellaria baicalensis Georgi and Radix paeoniae Alba on periodontitis mice and compare the results of the two herbs for the treatment of the periodontitis mice.

METHODSSixty-four SPF 12-week-old male Kunming mice were selected and randomly divided into four groups:Control group(C); Experimental periodontitis group (P):the peridontitis models in Kunming mice were prepared by wrapping silk ligature and inoculating with putative periodontopathic bacteria; Scutellaria baicalensis Georgi treatment group (SG): periodontitis was induced by the same method described above, the mice were gavaged with Scutellaria baicalensis Georgi; Radix paeoniae Alba treatment group (RG): periodontitis was induced by the same method described above, the mice were gavaged with Radix paeoniae Alba.Four mice were sacrificed at each time point of the end of 4, 6, 8 and 10 weeks in each group. The histopathological changes of periodontal tissue were observed under microscope with HE staining. The level of serum IgG1 and IgG2a was measured by enzyme-linked immunosorbent assay (ELISA) .

RESULTSA serious inflammatory response, alveolar progressive absorption and a large number of osteoclasts were observed in the experimental periodontitis group.However, in SG and RG, the inflammation of the periodontal tissue was decreased and tissue repair was significant. The level of serum IgG2a in SG (6 week:0.934 ± 0.006, 8 week:0.743 ± 0.009, 10 week: 0.674 ± 0.008) and RG (6 week: 1.023 ± 0.032, 8 week: 0.851 ± 0.032, 10 week:0.790 ± 0.009) was significantly decreased after the mice were gavaged with the two herbs(P < 0.01). The level of serum IgG2a in SG was significantly lower than that of RG (P < 0.01). The level of serum IgG1 in SG (6 week: 0.314 ± 0.006, 8 week: 0.344 ± 0.004, 10 week: 0.367 ± 0.006) and RG (6 week: 0.287 ± 0.005, 8 week: 0.303 ± 0.058, 10 week: 0.336 ± 0.006) were significantly increased (P < 0.01). The level of serum IgG1 in SG was significantly higher than that of RG (P < 0.01).

CONCLUSIONSBoth the aqueous extracts of Scutellaria baicalensis Georgi and Radix paeoniae Alba showed therapeutic effect on periodontitis in mice.Scutellaria baicalensis Georgi was more effective than Radix paeoniae Alba.

Aconitum ; Animals ; Immunoglobulin G ; metabolism ; Male ; Mice ; Paeonia ; Periodontitis ; metabolism ; Plant Extracts ; pharmacology ; Scutellaria baicalensis ; Water

OBJECTIVETo probe the risk of colorectal polyp, colon and rectal carcinoma and the intake of NSAIDs.

METHODSCase-control study participants were from patients who underwent colonoscopy at different hospitals, the persons with the above disease was as cases, and those without the above diseases was as controls. Use of NSAIDs was assessed by interviewing the participants with a questionnaire which include a list of NSAIDs and related dietary and life style factors and family history.

RESULTSThere are 37 cases of colorectal polyp, 105 cases of colon carcinoma and 142 cases of rectal carcinoma and 66 controls. Adjusted for potential confounders, the risk of colorectal polyposis, colon carcinoma and rectal carcinoma were markedly reduced by NSAIDs. The OR values were 0.21 (95% CI 0.07-0.65, P = 0.007), 0.13 (95% CI 0.05-0.35, P < 0.001), 0.15 (95% CI 0.11-0.58, P < 0.001) respectively. The risk of the above diseases were also reduced markedly by aspirin, the OR values were 0.265 (95% CI 0.07-0.96, P = 0.044), 0.10 (95% CI 0.03-0.35, P < 0.001), 0.15 (95% CI 0.04-0.49, P = 0.002) respectively. The risk of colon carcinoma was also reduced by profen, with the OR being 0.11 (95% CI 0.02-0.64, P = 0.014).

CONCLUSIONSAspirin and other NSAIDs could reduced the risk of colorectal polyp, colon carcinoma and rectal carcinoma markedly. Aspirin was the most prospective chemopreventive agents for colorectal polyp, colon and rectal carcinoma for its capability of reducing the risk of cardio-cerebral vascular disease as well.

Adult ; Anti-Inflammatory Agents, Non-Steroidal ; therapeutic use ; Aspirin ; therapeutic use ; Colorectal Neoplasms ; prevention & control ; Female ; Humans ; Ibuprofen ; therapeutic use ; Logistic Models ; Male ; Middle Aged ; Piroxicam ; therapeutic use ; Polyps ; prevention & control ; Rectal Neoplasms ; prevention & control ; Risk Factors ; Sulindac ; therapeutic use ; Time Factors

Parasitic diseases still remain the world's greatest health problems and cause huge economic burden in poor areas. The drugs currently used to treat protozoiases and helminthiases have certain defects, and it is urgent to develop more effective therapeutic drugs for these diseases. Berberine is one kind of important anti-inflammatory agents originally derived from Coptis rhizoma. The derivatives of berberine are obtained by modifying the structural site of berberine. In addition to its anti-inflammatory and anti-microbial activities, berberine and its derivatives also have significant anti-parasitic activity. In this paper, we summarized recent progress in the use of berberine and its derivatives against the infections of protozoa ( Leishmania spp ., Trypanosoma spp. , Toxoplasma gondii, Plasmodium falciparum, and Eimeria tenella) and helminths ( Schistosoma mansoni, Schistosoma. japonicum, Echinococcus granulosus, and Toxocara canis), which may providea useful reference for researchers in this field.

Objective:To investigate the therapeutic effect of artesunate on mice with acute Toxoplasma gondii ( T. gondii) infection. Methods:Based on body weight (16 - 18 g), sixty-four C57BL/6 female mice aged 6 - 8 weeks were divided into 4 groups by random number table method: uninfected control group without treatment; T. gondii-infected group (Tg group), each mouse was intraperitoneally (i.p.) infected with 100 tachyzoites of T. gondii RH strain; T. gondii-infected + artesunate treatment group (Tg + ART group), 3 hours after each mouse i.p. infected with 100 tachyzoites of T. gondii RH strain, artesunate solution was i.p. injected at a dose of 30 mg/kg, once a day for a total of 7 consecutive days; T. gondii-infected + sulfadiazine treatment group (Tg + SDZ group), 3 hours after each mouse i.p. infected with 100 tachyzoites of T. gondii RH strain, sulfadiazine solution was orally administrated at a dose of 100 mg/kg, once a day for a total of 7 consecutive days. There were 16 mice in each group, in which 10 mice were used to observe survival time and 6 mice were used to monitor body weight and collect tissue samples. Mice were weighed every day from day 1 post infection (p.i.); mice were sacrificed at day 7 p.i., the liver weights of mice were weighed and the liver indexes were calculated; liver tissues were paraffin-embedded, sectioned, and stained with hematoxylin-eosin (HE), and the pathological changes of liver tissues of mice in each group were observed under a light microscope. The expression levels of T. gondii major surface antigen 1 (SAG1) in the liver tissues of mice in each group were detected by real-time quantitative PCR for evaluating parasite load. Results:All mice in the uninfected control group were survived. The survival time was 7 - 9 days in Tg group, 8 - 11 days in Tg + ART group, and 9 - 13 days in Tg + SDZ group. Compared with Tg group, the survival times of mice in Tg + ART group and Tg + SDZ group were significantly longer ( P < 0.05). On day 7 p.i., compared with uninfected control group, Tg + ART group or Tg + SDZ group, the body weight of mice in Tg group was lower ( P < 0.05); however, there was no significant difference of body weight in Tg + ART group and Tg + SDZ group compared with uninfected control group ( P > 0.05). Compared with Tg group, Tg + ART group and Tg + SDZ group had lower liver indexes and SAG1 mRNA expression levels in the liver tissues ( P < 0.05 or < 0.001), and liver histopathological changes were milder. Compared with Tg + SDZ group, there was no significant difference in both liver index and SAG1 mRNA expression level in the liver tissue of Tg + ART group ( P > 0.05). Conclusion:Artesunate solution i.p. injection can prolong the survival time, reduce parasite load in the liver, and attenuate hepatic pathological damage, to a certain extent, of mice with acute T. gondii infection.