Source analysis of lymphocytes secreting interleukin-22 from spleen of mice infected with Trichinella spiralis at the early encapsulated stage / 中华地方病学杂志

Objective To investigate the cell origin of interleukin (IL)-22-secreting cell of mice infected with Trichinella spiralis (T.spiralis) at the early encapsulated stage.Methods Twelve Balb/c mice were divided into the infected group and the control group according to body weight by random number table.The infected mice were intragastrically administrated with 300 muscle larvae of T.spiralis,and the control mice were given the same amount of normal saline.The IL-22-secreting cell subsets in mouse splenic lymphocytes were detected by flow cytometry at the fourth week after infection.Results The proportion of IL-22-secreting cells in splenic lymphocytes of T.spiralis infected mice was increased when compared with control group [(0.88 ± 0.25)％ vs (0.28 ±0.17)％,t =-4.899,P ＜ 0.05].There was no significant difference between the proportion of CD3+IL-22+ cells and CD3-IL-22+ cells in the splenic lymphocytes of the infected group [(0.29 ± 0.17)％ vs (0.51 ± 0.17)％,t =-2.195,P ＞ 0.05],and the percentage of CD3-IL-22+ cells were similar between the infected group and the control group [(0.51 ± 0.17)％ vs (0.44 ± 0.22)％,t =-0.600,P ＞ 0.05].The proportion of CD3+IL-22+ cells in the infected group was significantly higher than that in the control group [(0.29 ± 0.17)％ vs (0.07 ± 0.06)％,t =-3.068,P ＜ 0.05],and the percentage of CD4+IL-22+ T cells and γδTCR+IL-22+ T cells were obviously increased in CD3+ lymphocytes [(1.28 ± 0.54)％ vs (0.16 ± 0.07)％,(0.33 ± 0.22)％ vs (0.02 ± 0.00)％,t =-4.997,-3.342,P ＜ 0.05].Conclusions The proportion of IL-22-secreting splenic lymphocytes is increased in mice infected with T.spiralis at the early encapsulated stage.The rise is caused by increased numbers of IL-22-secreting CD3 + lymphocytes,especially CD4+ T cells and γδT cells.

Role of spinal Toil-like receptor 4 signaling pathway in development of inflammatory pain in rats / 中华麻醉学杂志

Objective To evaluate the role of spinal Toll-like receptor 4 (TLR4) signaling pathway in the development of inflammatory pain in rats.Methods Thirty-six adult male Sprague-Dawley rats were divided into 3 groups (n=12 each) using a random number table:control group,inflammatory pain group and TLR4 signaling pathway inhibitor epigallocatechin gallate (EGCG) group (EGCG group).Inflammatory pain was induced by injecting 50 μl of complete Freund's adjuvant (CFA) into the ankle joint cavity of the left hindpaw of rats anesthetized with isoflurane.At 1-3 days after injection of CFA,EGCG 30 μg was intrathecally injected once a day in group EGCG.At 1,3 (30 min after intrathecal injection),5and 7 days after injection of CFA,the mechanical paw withdrawal threshold (MWT) and thermal paw withdrawal latency (TWL) were measured.The ipsilateral L4.5 segments of the spinal cord were removed at 3 days after CFA injcction for determination of TLR4 expression (by Western blot) and contents of tumor necrosis factor-alpha (TNF-oα),interleukin-6 (IL-6) and IL-1β in the spinal dorsal horn (by enzymelinked immunosorbent assay).Results Compared with control group,the MWT was significantly decreased and the TWL was shortened at each time point after injection of CFA,the expression of TLR4 in the spinal dorsal horn was up-regulated,and contents of TNF-α,IL-6 and IL-1β in the spinal dorsal horn were increased in inflammatory pain group (P＜ 0.05),and no significant change was found in the parameters mentioned above in EGCG group (P＞0.05).Compared with inflammatory pain group,the MWT was significantly increased and the TWL was prolonged at each time point after injection of CFA,the expression of TLR4 in the spinal dorsal horn was down-regulated,and contents of TNF-α,IL-6 and IL-1β in the spinal dorsal horn were decreased in EGCG group (P＜0.05).Conclusion Spinal TLR4 signaling pathway is involved in the development of inflammatory pain in rats.

Research progress in the third-generation genomic editing technology - CRISPR/Cas9 / 中华医学遗传学杂志

CRISPR/Cas9 technology originated from type II CRISPR/Cas system, which is widely found in bacteria and equips them with acquired immunity against viruses and plasmids. CRISPR-associated protein Cas9 is a RNA-guided endonuclease, which can efficiently introduce double-strand breaks at specific sites and activate homologous recombination and/or non-homologous end joining mechanism for the repair of impaired DNA. Features such as easy-to-use, cost-effectiveness, multiple targeting ability have made it the third-generation genomic engineering tool following ZFNs and TALENs. Here the history of discovery and molecular mechanism of the CRISPR/Cas9 technology are reviewed. The rapid advance in its various applications, especially for the treatment of human genetic disorders, as well as some concomitant problems are discussed.

Role of interleukin-12 in spinal cord in maintenance of arthritic pain in rats / 中华麻醉学杂志

Objective To evaluate the role of interleukin-12 (IL-12) in the spinal cord in the maintenance of arthritic pain (AP) in rats.Methods Adult male Sprague-Dawley rats, aged 6-8 weeks,weighing 200-300 g, were randomly divided into 4 groups using a random number table: control group (group C, n=6);AP group (n=9);phosphate buffer solution (PBS) group (n=6);IL-12 antibody group (n =6).AP was induced by injecting 50 μl of complete Freund' s adjuvant into the ankle joint cavity of the left hindpaw of rats anesthetized with isoflurane.Goat anti-rat IL-12 antibody 1.50 μg (20 μl) was intrathecally injected on 9 days after establishment of the model in group IL-12 antibody, while 0.01 mol/L PBS (20 μl) was administered in group PBS.The mechanical paw withdrawal threshold to yon Frey filament stimulation (MWT) was measured before establishment of the model (baseline) and on 9 and 10 days after establishment of the model.The rats were sacrificed after the last measurement of pain threshold, and the L4 6 segments of the spinal cord were obtained for detection of IL-12 expression in the spinal dorsal horn (by immunofluorescence) , and the co-expression of IL-12 and glial fibrillary acidic protein (an astrocyte marker) was examined simultaneously in group AP.Results Compared with group C, the MWT was significantly decreased, and the expression of IL-12 was up-regulated on 9 and 10 days after establishment of the model in AP, PBS and IL-12 antibody groups.Compared with group AP, the MWT was significantly increased at 10 days after establishment of the model, and the expression of IL-12 was down-regulated in group IL-12 antibody, and no significant change was found in the MWT at 10 days after establishment of the model and expression of IL-12 in group PBS.IL-12 was co-expressed with glial fibrillary acidic protein in group AP.Conclusion IL-12 in the spinal cord is involved in the maintenance of AP in rats.