Objective:To study the optimal water extraction method for Shugan Xiaocuo granules. Methods: The study was car-ried out according to the orthogonal tablet. The water amount, extraction time,extraction times and soak time as the influencing factors, and the extractive yield and baicalin content as the indices in a comprehensive evaluation, the optimal extraction technology was screened. Results:The optimal extraction technology was as follows:soaked 30 minutes before cooking, 8-fold water,extracted for 3 times with 1h for every time. Conclusion:The verification test showed that the optimum extraction process was stable,reasonable and feasible.

Objective:To optimize the extraction technology of volatile oil in myrrh. Methods: Using the extraction quantity of volatile oil as the index, the volatile oil was extracted by an essential oil extractor, and U6 (64 ) uniform design was used to investigate the influencing factors such as the immersion time ( A) , the mesh number ( B) , the extraction time ( C) and the additive water amount ( D) . Results:The optimized extraction technology was as follows: myrrh was crushed to obtain 10 mesh powder, soaked for 10 mi-nutes with 8-fold amount of water, and then extracted 3 hours. Conclusion:The quantity of volatile oil extracted from myrrh is accu-rate, the technology is reasonable, and the research provides scientific basis for the extraction of volatile oil in myrrh.

OBJECTIVE:To optimize water reflux extraction technology of Jiuwei yiwei mixture. METHODS:With the over-all score of the content of salvianoli acid B,a water-soluble component of the principal drug Salvia miltiorrhiza,the content of pae-oniflorin in Paeonia Radix Rubra and dry ointment yield rate as the observed index,and taking soaking time,extraction time, amount of water added,and extraction times as the factors,L9(34)orthogonal test was designed to optimize the extraction technolo-gy of Jiuwei yiwei mixture. And the verification test was conducted. RESULTS:The optimal technology was to add water with an amount 8 times as much as that of the drugs to soak them for 1 h and to conduct extraction twice,1 h for each. The overall scores of 3 batches of samples in the verification test were respectively 98.793,99.078 and 99.220(RSD<2%,n=3). CONCLUSIONS:The optimal technology for Jiuwei yiwei mixture is stable and feasible.

BACKGROUND:The application of virtual reality technology in preoperative simulation can reduce the risk in operation effectively and improve the quality of surgery. Virtual reality technology has very important practical significance in the application of surgery. OBJECTIVE:The reverse engineering and virtual reality technology were used to achieve the preoperative simulation of a case of Pilon fracture. METHODS:The affected bones were reconstructed according to the CT data using the patient’s ankle portion in MIMICS software, and the separated bone fractures were restored. According to the characteristics of virtual reality technology, further processing on bone model was carried out using Geomagic software;the models of the fractured bones were exported with STL format. The restored bone fragments were checked up to determine integrity in the virtual reality operation platform. These models were used to do simulated operations in the virtual reality operation platform. RESULTS AND CONCLUSION:A case of Pilon fracture was simulated by using reverse engineering and virtual reality technology preoperatively. The processed 3D model was introduced into the virtual reality system to simulate the operation and help doctors choose the type, position and direction of the surgical approach and plate;the effect is good.

Objective:To optimize the ethanol reflux extraction process for Aitong cataplasm .Methods: Orthogonal design was used to investigate the effect of solid-liquid ratio,extraction time and times on the extraction technology of Aitong cataplasm with the content of asarinin and dry extract yield as the indices .Results:The best extraction conditions were as follows:extracted 3 times with 8-fold amount of 70%ethanol, and 1 hour for each time.Conclusion: The optimized extraction process is stable and feasible , which can be used to extract Aitong cataplasm .

OBJECTIVE:To determinate the release rate and in vitro transdermal rate of asarinin in Cancer pain cataplasm. METHODS:Using homemade devices and modified France diffusion,isolated skin of rats as barrier,normal saline as solvent,the content of asarinin was determined by HPLC. Release rate of Cancer pain cataplasm within 20,50,80 and 120 min and transder-mal amount within 2,4,8,12,24 h were investigated,and accumulative release rate and accumulative transdermal rate were cal-culated. RESULTS:Accumulative release rate by 120 min of asarinin in Cancer pain cataplasm was 73.01%;24 h in vitro transder-mal rate was 26.01%,and transdermal kinetics equation of asarinin was Q=5.717 7t1/2-0.385 4(r=0.979). CONCLUSIONS:Cancer pain cataplasm has good release and transdermal performance. Its transdermal kinetics is in line with Higuchi equation.

Objective To optimize the supercritical CO2extraction technology condition of radix cynanchi paniculati. Methods The paeonol extract was used as indexes.The content of paeonol was determined by high performance liquid chromatography method.The extraction condition was optimize tby design-response surface method. Results The optimum process conditions for the extraction pressure was 10.81 MPa,extraction temperature was 55 ℃,flow rate of CO2was 59.91 L·h-1,extraction time was 2.42 h. Conclusion The content of paeonol with optimum extraction process of radix cynanchi paniculati is high,process is stable and feasible.

Objective To evaluate the clinical effects of intra-articular injection of autologous adipose derived stem cells (ADSCs) or ADSCs combined with hyaluronate acid (HA) for knee osteoarthritis.Methods From May 2013 to May 2015,a total of 108 patients with knee osteoarthritis (Kellgren-Lawrence grades:1-3) were recruited in the present study.The patients were randomly divided into three groups:ADSCs,HA and ADSCs+HA.All patients (36 cases in each group) were injected with the drug in the joint cavity once a week for three weeks.The methods used for evaluating the clinical manifestations and joint damage on MRI included visual analogue scale (VAS),Western Ontario and McMaster University (WOMAC) osteoarthritis index and whole-organ magnetic resonance imaging score (WORMS).Evaluations were conducted before injection and at 3,6,12,24 and 36 months after injection.Results All patients were followed up for 36 months without any dissociation.No adverse reaction was observed during the treatment and at the follow-up duration.The VAS score and WOMAC total score of the ADSCs group and the ADSCs+HA group were better than those in the HA group at each time point after injection (P＜0.05).The average VAS of the ADSCs group decreased from 4.14±1.42 at pre-injection to 2.39±1.74 at 36 months after injection.The WOMAC total score decreased from 42.86± 31.24 to 27.17±27.99.The average VAS in the ADSCs+HA group decreased from 4.25±1.13 to 2.31±1.74,and the WOMAC total score decreased from 34.92±22.62 to 21.33±21.38.However,the average VAS and WOMAC total score of the HA group at 36 months after injection were higher than those before the injection.In terms of the VAS at 3 months after injection,the ADSCs+HA group scored better than that of the ADSCs group (P＜0.05).There was no significant difference in WOMAC scores between the ADSCs and ADSCs+HA groups at each time point after injection (P＞0.05).The WORMS cartilage injury score improved in 10 patients with ADSCs after injection (P＜0.05).The subchondral bone wear score improved as well (P＜0.05).The difference of WORMS cartilage injury scores before and after injection was correlated with the difference of the WOMAC total score (r=0.790,P=0.007) and that of VAS score difference (r=0.800,P=0.005).Conclusion Autologous ADSCs and ADSCs combined with HA intra-articular injections can effectively relieve pain and improve function of patients with knee osteoarthritis for 36 months.In addition,ADSCs combined with HA injections can relieve pain more effectively within a short duration.Cartilage repair is associated with joint function improvement.

Objective: To investigate the feasibility of myeloid and plasmacytoid dendritic cell combined vaccines loaded with heat-treated Lewis lung cancer cell lysates for treatment of lung cancer in mice. Methods: Bone marrow cells were induced by the recombinant mouse fms-like tyrosine kinase receptor 3 ligand (rmFlt3-L) in vitro, myeloid dendritic cells (mDC) and plasmacytoid dendritic cells (pDC) were separated by magnetic beads. The mDC, pDC, and mDC∶pDC=1∶1 were stimulated with heat-treated Lewis lung cancer cell lysates, respectively. The effects of each group on stimulating of lymphocyte proliferation and inducing of T cell to kill tumor cells in vitro were compared. The alternations of the immunophenotypes of CD80, CD86, CD40 and major histocompatibility complex Ⅱ (MHC-Ⅱ) were detected by flow cytometry. The secretion of cytokines including interlukin-12 (IL-12), interlukin-6 (IL-6), and tumor necrosis factor α (TNF-α) were detected by enzyme-linked immunosorbent assay (ELISA). Results: The lymphocyte proliferation in mice stimulated with mDC+ pDC group loaded with heat-treated Lewis lung cancer cell lysates was 10.80±0.66, significantly higher than 8.63±0.65 of mDC group and 7.10±0.46 pDC group under the same culture conditions, respectively (P<0.05). When the ratio of effector cells: target cells (E∶T) was 10∶1, the killing rate of the mDC+ pDC group loaded with heat-treated tumor cell lysate was 31.68%±2.93%, significantly higher than 17.44%±0.97% of mDC group and 10.29%±1.33% of pDC group, respectively (P<0.05). When the ratio of E∶T was 20∶1, the killing rate of the mDC+ pDC group loaded with heat-treated tumor cell lysate was 54.77%±3.28%, significantly higher than 35.25%±1.51% of mDC group and 15.52%±0.73% of pDC group, respectively (P<0.05). When the ratio of E∶T was 40∶1, the killing rate of the mDC+ pDC group loaded with heat-treated tumor cell lysate was 73.01%±0.91%, significantly higher than 51.36%±0.58% of mDC group and 22.65%±1.28% of pDC group, respectively (P<0.05). With the rate of E∶T increased, the killing rate also increased. The mean fluorescence intensities of surface molecules including CD80, CD86, CD40 and MHC-Ⅱ of mDC: pDC=1 group pulsed with heat-treated Lewis lung cancer cell lysates were higher than those of mDC group and pDC group. The IL-6 cytokine concentrations of mDC+ pDC group, mDC group and pDC group loaded with heat-treated Lewis lung cancer cell lysates were (586.67±52.52) pg/ml, (323.33±67.14) pg/ml and (166.67±16.07) pg/ml, respectively. The concentrations of IL-12 in each group were (2 568.75±119.24) pg/ml, (2 156.25±120.55) pg/ml and (672.92±31.46) pg/ml, respectively. The concentrations of TNF-α in each group were (789.33±48.08) pg/ml, (584.89±116.49) pg/ml and (291.56±40.73) pg/ml, respectively. The concentrations of IL-6, IL-12 and TNF-α secreted by mDC+ pDC group were much higher than those of mDC group and pDC group under the same culture conditions (P<0.05). Conclusions The mDCs and pDCs combined vaccines pulsed with heat-treated Lewis lung cancer cell lysates have synergistic effects on inducing of T lymphocyte proliferation and killing tumor cells in vitro. This synergistic anti-tumor effect is related with up-regulation of co-stimulatory molecules and increased secretion of cytokines.

To investigate the feasibility of myeloid and plasmacytoid dendritic cell combined vaccines loaded with heat?treated Lewis lung cancer cell lysates for treatment of lung cancer in mice. Methods Bone marrow cells were induced by the recombinant mouse fms?like tyrosine kinase receptor 3 ligand ( rmFlt3?L) in vitro, myeloid dendritic cells ( mDC) and plasmacytoid dendritic cells (pDC) were separated by magnetic beads. The mDC, pDC, and mDC ∶ pDC＝1 ∶ 1 were stimulated with heat?treated Lewis lung cancer cell lysates, respectively. The effects of each group on stimulating of lymphocyte proliferation and inducing of T cell to kill tumor cells in vitro were compared. The alternations of the immunophenotypes of CD80, CD86, CD40 and major histocompatibility complex Ⅱ( MHC?Ⅱ) were detected by flow cytometry. The secretion of cytokines including interlukin?12 (IL?12), interlukin?6 (IL?6), and tumor necrosis factor α ( TNF?α) were detected by enzyme?linked immunosorbent assay ( ELISA). Results The lymphocyte proliferation in mice stimulated with mDC+pDC group loaded with heat?treated Lewis lung cancer cell lysates was 10.80±0.66, significantly higher than 8.63±0.65 of mDC group and 7.10±0.46 pDC group under the same culture conditions, respectively ( P＜0.05). When the ratio of effector cells:target cells (E ∶ T) was 10 ∶ 1, the killing rate of the mDC+pDC group loaded with heat?treated tumor cell lysate was 31.68%±2.93%, significantly higher than 17.44%±0.97% of mDC group and 10.29%±1.33% of pDC group, respectively (P＜0.05). When the ratio of E ∶ T was 20 ∶ 1, the killing rate of the mDC+pDC group loaded with heat?treated tumor cell lysate was 54.77%± 3.28%, significantly higher than 35.25%± 1.51% of mDC group and 15.52%±0.73% of pDC group, respectively (P＜0.05). When the ratio of E ∶ T was 40 ∶ 1, the killing rate of the mDC+pDC group loaded with heat?treated tumor cell lysate was 73.01%± 0.91%, significantly higher than 51.36%± 0.58% of mDC group and 22.65%± 1.28% of pDC group, respectively (P＜0.05 ). With the rate of E ∶ T increased, the killing rate also increased. The mean fluorescence intensities of surface molecules including CD80, CD86, CD40 and MHC?Ⅱ of mDC:pDC＝1 group pulsed with heat?treated Lewis lung cancer cell lysates were higher than those of mDC group and pDC group. The IL?6 cytokine concentrations of mDC+pDC group, mDC group and pDC group loaded with heat?treated Lewis lung cancer cell lysates were (586.67±52.52) pg／ml, (323.33±67.14) pg／ml and (166.67± 16.07) pg／ml, respectively. The concentrations of IL?12 in each group were ( 2 568.75± 119.24) pg／ml, (2 156.25±120.55) pg／ml and (672.92±31.46) pg／ml, respectively. The concentrations of TNF?α in each group were (789.33±48.08) pg／ml, (584.89±116.49) pg／ml and (291.56±40.73) pg／ml, respectively. The concentrations of IL?6, IL?12 and TNF?α secreted by mDC+pDC group were much higher than those of mDC group and pDC group under the same culture conditions ( P＜0.05). Conclusions The mDCs and pDCs combined vaccines pulsed with heat?treated Lewis lung cancer cell lysates have synergistic effects on inducing of T lymphocyte proliferation and killing tumor cells in vitro. This synergistic anti?tumor effect is related with up?regulation of co?stimulatory molecules and increased secretion of cytokines.