Objective:To study effect of tripterygium glycosides(TG)on T-lymphocyte subsets in rats with idiopathic thrombocytopenic purpura(ITP),and further to discuss the possible mechanism of TG in treating ITP.Methods:ITP animal model was established by intraperitoneal injection of exogenous anti-platelet–serum(APS).Sixty model rats were randomly divided into five groups:normal group,saline group,the high and low dosage of TG groups,prednisone group.Every rat was giving liquid medicine by intragastric administration one time per day for ten days 36 hours after the first intraperitoneal injection of exogenous APS.The changes of peripheral hemogram,the amount of megakaryocyte of bone marrow and T-lymphocyte subsets were observed.Results:In high dosage of TG group,the number of platelet count increased obviously,the number of megakaryocyte in bone marrow,CD8+and CD4+decreased,and CD4+/CD8+increased.Conclusion:High dosage of TG had obviously therapeutic effect on IPT rat,such as increasing platelet count obviously,reducing number of megakaryocyte in bone marrow obviously and ameliorating T-lymphocyte subsets.The mechanism may be related to regulating the cell immune function.

Objective To establish a stable animal model for sequentially dynamic and direct monitoring of the skin wound infection. Methods The mice with full-thickness skin incisions were replicated. After immediate subcutaneous suture,the mice were randomly divided into four groups,ie,Group A was inoculated with 50 μl sterile PBS solution),Groups B,C and D were inoculated with 50 μl suspension containing 1 × 106,1 × 108 and 1 × 1010 colony forming unit (CFU)/ml bioluminescent methicillin-resistant staphylococcus aureus (MRSA) respectively.Then,the diet behavior of each group was observed and the mean weight and mortality of each group were also recorded at different time points.The bioluminescent intensity of fluoresce in the wounds was recorded at different time points by using the charge-coupled device (CCD) based imaging system.Local wound tissues were incised at 24 hours after inoculation for HE staining so as to observe wound inflammatory reaction in each group.Wound healing time of each group was also recorded. Results ( 1 ) Average weight:Groups A and B showed unobvious changes in weight; Group C lightened until day 3 after inoculation and then recovered gradually to the preinoculation level at day 14; Group D lightened gradually until death.(2)Mortality:Groups A and B had no death; Group C had 10％ deaths at day 14; Group D had 100％ deaths.(3) Bioluminescent intensity of wounds:Groups A and B showed a gradual weakened luminescence since the day of inoculation and had almost complete disappearance at days 5 and 7 respectively; there was no sign of obvious increase or decrease in Group C from the day of inoculation till day 14 ; Group D had a gradual increase since the day of inoculation and the luminous area expanded until the death.(4) HE staining at 24 hours after inoculation:all the four groups showed inflammatory cell infiltration,especially in Groups C and D.(5) Wound healing time:wound healed at days 5 and 7 after inoculation in Groups A and B; the wounds showed no healing even at day 14 in the Group C,but the wounds length and area did not show obvious enlargement or diminishment ; the wounds extended gradually until the death in the Group D,since the day of inoculation. Conclusions The inoculation of 50 μl suspension with 1 × 108 CFU/ml bioluminescent MRSA to full-thickness skin incision rats allows direct,real-time dynamic and continuous detection of the occurrence and development of the wound infections.The infection model is easy to make and has stability and high repeatability.

Objective To study the influence of serum containing Fuzheng Toudu Qudu Recipe, a Chinese formula with the actions of supporting healthy qi to expel and remove toxicity, on serum levels of interleukin 2 (IL-2) and soluble interleukin 2 receptor (sIL-2R) at different stages of CD34+derived dendritic cells (DC) of patients with minimal residual disease of myelogenous leukemia ( MRD-L) , and to explore the biological mechanism of Fuzheng Toudu Qudu Recipe in promoting CD34+ to transform into DC in MRD-L patients. Methods Bone marrow mononuclear cells ( BMMC) were separated from the bone marrow of acute myeloid leukemia patients at complete remission stage by using Ficoll centrifugation. CD34+ cells were isolated by using immuno-magnetic mircobeads method, and then were cultured with various concentrations of Chinese medicine medicated serum and cytokines in vitro for the induction of DC. The morphologic characteristics of DC were observed with the inverted phase contrast microscope, and the expression levels of DC surface molecules such as CD83, CD80, CD86, CD1a and HLA-DR were detected by using flow cytometry. On culturing day 0, 6 and 9, serum levels of IL-2 and sIL-2R of each group were measured by enzyme-linked immunosorbent assay ( ELISA). Results ( 1) Chinese medicine medicated serum combined with cytokines was effective on promoting CD34+ to differentiate into DC with typical morphology, and inducing DC to have high expression of CD80, CD83, CD86 and HLA-DR, which differed from those in fetal calf serum (FCS) group and blank rabbit serum group (P<0.01). Middle- and low-dose combination groups increased expression of CD1a, which differed from high-dose combination group and cytokines group ( P<0.01). ( 2) Content of IL-2 in combination groups was higher than that in blank rabbit serum group on culturing day 0. In the combination groups, IL-2 was higher on culturing day 6 and 9 than that on culturing day 0. Middle and high-dose combination groups had higher IL-2 content on culturing day 9 than on culturing day 6 ( P<0.05 or P<0.01). At the same time point, combination groups had higher IL-2 content than the blank rabbit serum group (P<0.05 or P<0.01). (3) Content of sIL-2R in the combination groups was lower than that in blank rabbit serum group on culturing day 0. In the combination groups, sIL-2R was lower on culturing day 9 than that on culturing day 0 and 6 ( P<0.01) . High-dose combination group had lower sIL-2R content on culturing day 6 than that on culturing day 0 (P<0.05), and the difference of sIL-2R in other groups was insignicant between on culturing day 6 and on culturing day 0 ( P>0.05) . At the same time point, combination groups had lower IL-2 content than the blank rabbit serum group (P<0.05 or P<0.01). Conclusion Fuzheng Toudu Qudu Recipe is effective on increasing serum content of IL-2 and reducing sIL-2R content, and the changes of cytokine contents are more obvious along with the maturity of DC, which indicates that the recipe plays positive effect in the process of promoting CD34+cells to differentiate into DC.

Objective: To explore the safety and feasibility of colonoscopy - assisted transanal minimally invasive surgery via glove port (CA-TAMIS-GP) in the treatment of early rectal tumors. Methods: A total of 67 patients evaluated as early rectal tumors (adenoma limited within mucosal layer) with diameter ≤4.0 cm at Department of Anal-Colorectal Surgery, Hangzhou Third People′s Hospital from July 2013 to March 2017 were prospectively enrolled in the study. Benign tumors were diagnosed by preoperative imaging in all the patients with the distance to anal edge of 4 to 20 cm. Patients were randomly divided into treatment group and the control group according to the random number table. The treatment group (n=32) underwent CA-TAMIS-GP, including 19 males and 13 females with mean age of (55.6±11.2) years and mean tumor size of (3.3±0.4) cm, while the control group (n=35) underwent endoscopic submucosal dissection (ESD, control group), including 20 males and 15 females with mean age of (52.9±12.3) years and mean tumor size of (3.4±0.5) cm. Differences of baseline data between two groups were not significant (all P>0.05). The specific method of CA-TAMIS-GP was as follows: a surgical rubber glove sleeve (No.6) was passed through the anal device; the glove was fixed at the anvil device; after fully expanding the anus, the anal sac was placed into the anus with the fingers outside; then, the cuff and the anal sac were sutured and fixed to the perianal; a well-tight glove path was established; the ultrasonic scalpel, grasper and the colonoscopy lens connected to the host platform and the electric negative pressure suction were inserted into the three finger sleeves respectively and fixed by rubber band or silk thread; the laparoscopic instruments such as the grasper and the ultrasonic scalpel were used for pulling, grasping, cutting, electrocoagulation, suturing and other operations to complete the resection of rectal lesions. Efficacy, postoperative complication and operative cost, etc. between two groups were compared using the student′s t test, chi-square tests, and Fisher′s exact test. Results: Operations of two groups were completed successfully without conversion to laparotomy. Histopathologic examination showed all specimens had negative margins with the surgical resection of the layer to the submucosa, and showed no significant differences between two groups (P>0.05). Compared to the control group, the operation time was shorter [(49.5±14.6) minutes vs.(66.1±17.6) minutes, t=-4.235, P<0.001], and the intraoperative hemorrhage was less [(4.2±1.6) ml vs. (6.2±2.1) ml, t=-4.349, P<0.001] in the treatment group with significant differences. In the treatment group, 6 patients had mild anal pain or discomfort after operation, and 1 patient in the control group showed anal foreign body sensation. The difference was statistically significant [18.8% (6/32) vs. 2.9% (1/35), P=0.048]. The incidence of postoperative hematochezia in the treatment group was lower than that in the control group [9.4% (3/32) vs. 20.0% (7/35), P=0.310] without significant difference. The cost of consumables in the treatment group was (1586.9±204.4) yuan, which was lower than (7694.4±1123.2) yuan in control group, and the difference was statistically significant (t=-30.880, P<0.001). All the patients were followed up for 6 to 36 months after operation, and no recurrence or long-term complication occurred in the treatment group, while 1 case developed local recurrence in the control group. Conclusion CA-TAMIS-GP is a safe and effective method for early rectal tumors with simple and economical characteristics, which broadens the application of colonoscopy.

OBJECTIVE To prepare Xiongzhi shigao decoction soluble microneedles， characterize it and investigate its transdermal properties in vitro. METHODS Two-step centrifugal method was used to prepare Xiongzhi shigao decoction soluble microneedles. The formability and mechanical property of the microneedles were evaluated from aspects of stroma fluidity， microneedle formability， needle hardness， etc. The appearance， mechanical strength， dissolution performance， skin barrier recovery performance and drug loading of the prepared microneedles were characterized by using active components of the soluble microneedle （chlorogenic acid， ferulic acid， notopterol， imperatorin， ligustilide， isoimperatorin） as indicators. The in vitro transdermal performance was investigated by Franz diffusion cell. RESULTS The soluble microneedle tips of Xiongzhi shigao decoction prepared in this study were conical， evenly distributed and of the same thickness， with good mechanical properties； the tip of the needle could be almost completely dissolved after being penetrated into the skin of rats for 2 hours， and the skin barrier recovery performance was good； the drug loading of chlorogenic acid， ferulic acid， notopterol， imperatorin， ligustilide and isoimperatorin were （87.04±1.12）， （67.69±1.23）， （20.65±0.17）， （35.00±0.11）， （153.83±0.21） and （23.52±0.50） μg per patch respectively. The results of in vitro transdermal study showed that cumulative release rates of 6 active components in this microneedle after 72 hours were 36.94%， 56.72%， 19.36%， 57.98%， 11.06% and 35.19%， respectively. CONCLUSIONS Xiongzhi shigao decoction soluble microneedles are prepared successfully in this study and have good formability， mechanical properties and pliable backing， which can significantly promote the transdermal drug delivery.