Cytokine-induced killer cells (CIK) is the fourth largest cancer treatment after surgery,chemotherapy and radiotherapy,and it is the development direction of cancer treatment.It is a new type of immune cell,and it is named after natural killer cell samples T lymphocytes as it express CD3 and CD56.Currently,CIK treatment has a broader range of clinical applications,and it has achieved the better clinical efficacy in the blood system cancer and solid tumors,The CIK adoptive immunotherapy is considered to be a new hope for the anticancer treatment.

Objective To study the effects of bone marrow mesenchymal stem cells(BMSCs) transplantation combined with artesunate(Art) on rat model of liver fibrosis.Methods 20 rats from 120 SD rats were selected randomly as normal control group.10 rats from those were executed and extracted marrow to culture BMSCs.The other 90 rats were injected intraperitoneally with CCl4 to make liver fibrosis rat models.10 rats from the 90 rats were selected randomly to ensure the liver fibrosis model was made successfully after 10 weeks.Then,the rest 80 rats were divided randomly into 4 groups:model group,Art treated group,BMSCs transplantation group and Art + BMSCs transplantation group.Fibrous degeneration of the hepatic tissue was detected by Masson stain.The mRNA expression of α-SMA,TGF-β1 in liver were detected by RT-PCR.The hydroxyproline(Hyp) level in liver was detected by enzyme digestion method.Serum TNF-αand TGF-β1 were detected by ELISA.Serum hepatic fibrosis parameters were detected by chemiluminescence and serum liver functional parameters were detected by biochemistry method.Results Liver functional parameters of Art + BMSCs transplantation group were better than those of the other groups.ALB in Art + BMSCs transplantation group was (29.13 ± 3.48) g/L,which was significantly higher than (26.31 ± 2.42) g/L in Art treated group (24.35 ±3.12)g/L in BMSCs transplantation group and (20.14 ±2.86) g/L in model group.There were significant differences among the 4 groups(q =6.27 ～ 19.01,all P ＜ 0.05).Liver fibrosis parameters of Art + BMSCs transplantation group were better than other groups.Hyp in Art + BMSCs transplantation group was (1.03 ±0.16) μg/L,which was significantly lower than (1.56 ± 0.19) μg/L in Art treated group,(1.45 ± 0.24) μg/L in the BMSCs transplantation group and (3.57 ±0.91) μg/L in the model group.There were significant differences among the 4 groups (q =20.47 ～ 27.70,all P ＜ 0.05).The expression of TGF-β1 mRNA of Art + BMSCs transplantation group was lower than other groups.The index of TGF-β1 mRNA expression in Art + BMSCs transplantation group was (20.16 ± 5.21),which was lower than (58.15 ±9.16) in Art treated group,(46.31 ±6.82) in the BMSCs transplantation group and (97.08 ± 14.23) in the model group.There were significant differences among the 4 groups (q =20.35 ～ 44.93,all P ＜ 0.05).Conclusion There're synergistic effects between Art and BMSCs in the treatment of liver fibrosis.

Objective To study two new factor Ⅸ mutations Cys82Ser and Ile288Ser in vitro and research the molecular mechanism of haemophilia B. Methods PcDNA3. 1 ( - ) FⅨwt expression plasmid was prepared. The mutated FⅨcDNA expression plasmids, PcDNA3.1 ( - ) FⅨM1 (Cys82Ser) and PcDNA3. 1 ( - ) F Ⅸ M2 (Ile288Ser) were constructed by megaprimer method respectively. Transient expression experiments were performed using HEK293 cells transfected with the expression vectors containing the wild-type or the mutation recombinant cDNA. PcDNA3. 1 ( - ) was used as a blank control. The expression proteins were detected by ELISA, factor activity assay and flourescence stain. Results The results suggested that the two FⅨ gene mutations did not induce the reduction of the mutant FⅨ mRNA compared with the wild-type FⅨ mRNA. The FⅨ:Ag in culture media and cell lysate of wild type conduct were assigned as 100. 0%. The results of PcDNA3.1 ( - ) FⅨ M1 mutation protein were (27. 1 ± 5. 2)% and (99.4 ±4. 1)% respectively. For PcDNA3. 1( - )FⅨM2, the results were (5.3 ± 1.8)% and (31.7 ±2. 5)% respectively. The FⅨ: C in culture media of wild type conduct was also assigned as 100. 0%. Then the two types of mutant protein were ( 8. 5 ± 3.2 ) % and ＜ 1%, respectively. Immunofluorescence microscopy result suggested that the intensity of perinuclear spot was reduced in cells transfected with PcDNA3.1 ( - ) FⅨM2 while staining for PcDNA3. 1 ( - ) FⅨM1 was predominantely diffuse without perinnclear enhancement. Conclusions These results strongly suggest that the FⅨ Cys82Ser mutation protein is not been correctly folded, by any possibility. The mutation protein has secretion defect. The secretion dysfunction and the protein degradation intracellular are possiblely the molecular pathology of Ile288Ser mutant protein.

The aim of the study is to evaluate the feasibility and safety of Bispectral index (BIS) monitoring in pediatric radio frequency catheter ablation. One hundred and six children aged 0. 6-12 years, scheduled for radio frequency catheter ablation, were randomly divided into two groups. In group A patients received BIS monitoring during the operation (n = 50), and the group B received modified Observer's Assessment of Alertness/Sedation (OAA/S) scaling (n = 56). The anesthesia was maintained with propofol target-controlled infusion. The intraoperative propefol target concentration was adjusted to maintain the BIS values between 55-65 in group A and OAA/S scale about 1 in group B respectively, The heart rate (HR), mean arterial pressure (MAP) and pulse oximetric saturation (SpO2) were measured before anesthetic induction, 1 min after induction, catheter puncturing and the end of operation respectively. The requirements of propofol, the times of supporting ventilation and recovery, the respiratory depression, nausea and vomiting postoperatively were also recorded. The intraoperative HR, MAP and SpO2 showed no differences between two groups, but the requirements of pmpofol, the times of supporting ventilation and recovery were less in group A than that of group B (P<0.05). All children didn't have nausea, vomiting and respiratory depression. The results suggest that in pediatric radio frequency catheter ablation, BIS monitoring has the advantages of timely adjustment of anesthetic depth, reducing anesthetic requirements, shortening the time of recovery, so as the perioperative safety can be improved.

Objective To study the in vitro culture condition for melanoblasts from human foreskin tissue. Methods The skin tissue taken from foreskin of children was treated with 0.5% dispase Ⅱ to separate epidermis from dermis, then with trypsin to obtain single cell suspension, which was cultured in modified medium for melanoblasts, i.e., MCDB254 medium supplied with several cell growth factors. Finally, melanoblasts were obtained based on the difference of adhesion speed. The morphology and proliferation of cultured melanoblasts were observed under a light microscope. DOPA staining, immunostaining with anti- S-100 and -tyrosinase related protein 2 (TRP2) antibodies, and transmission electron microscopy were per- formed to identify the cultured melanoblasts. Results The cultured human melanocytes displayed a match-like shape, scattered arrangement, syrmnetric double poles, slim cell body, highly refractive nuclei; meanwhile, the melanoblasts exhibited plentiful cytoplasm, large volume, bipolar or irregular shape and clonal growth. Additionally, the melanocytes were positive for TRP2, S-100 and Dopa staining, while the melanoblasts were positive only for TRP2. Electron microscopy revealed the presence of mature melanin granules (stage Ⅲ-Ⅳ ) in melanocytes but immature melanin granules (stage Ⅰ ) in melanoblasts. Conclu- sion Stable pure culture of melanoblasts has been realized with the reformed medium, which may lay a foundation for the investigation into the mechanism of epidermal pigmentation.

Lung distension belongs to chronic obstructive pulmonary disease (COPD) in modern medicine.In recent years,its disease incidence and mortality have increased year by year.Supported by the traditional Chinese medicine (TCM) standardization of the State Administration of TCM project-TCM preventive treatment practice guidelines-prevention of lung distension recrudescence,the first draft of the TCM Preventive Treatment Practice Guideline on Prevention of Lung Distension Recrudescence was formulated on the basis of preventive treatment theory,evidences from both ancient and modern literatures,and through the Delphi method as well as expert consensus conference.And then,peer review comments were collected,which laid the foundation for the further improvement of this guideline.This guideline was aimed to achieve the industry consensus,to improve the quality of life (QoL) in patients of the stable period,to reduce the economic burden of patients,as well as to improve both the theoretical connotation and practical value of preventive treatment in TCM.

Objective To test synchronization of cardiac mechanical contraction by means of advanced echocardiography and investigate the correlation of left ventricular ejection fraction (LVEF) and the indexes of mechanical dyssynchrony, and the relationship between DTI, STI and RT-3DE.Methods Control group (20 cases), chronic heart failure with a widened QRS complex (12 cases) and chronic heart failure with a shortened QRS duration (10 cases) were selected. We evaluated mechanical dyssynchrony with the DTI, the STI and the RT-3DE, and analyzed the correlation between the improvement degree of cardiac function and indexes of mechanical dyssynchrony, and the correlation between DTI, STI and RT-3DE. Results (1) In CHF groups (including shortened QRS group and widened QRS group), the indexes of synchronization of cardiac mechanical contraction were higher than control group ( <0.05) . (2) However, the indexes of mechanical dyssynchrony before operation showed no statistically significant difference between the widened QRS group and the shortened QRS group (>0.05) . (3) In CHF groups (including shortened QRS group and widened QRS group), the indexes of mechanical dyssynchrony before operation were higher more than after operation ( <0.05) . (4) In postoperation, the indexes of mechanical dyssynchrony showed no statistically significant difference between the widened QRS group and the shortened QRS group ( >0.05) . (5) There was a significant negative correlation between the LVEF and the indexes of mechanical dyssynchrony (<0.01) . (6) .In the indexes of synchronization of cardiac mechanical contractions, there are significant positive correlations between the DTI, the STI and the RT-3DE ( <0.01) . Conclusion Echocardiography can be used to screen CHF patients,and patients with left ventricular synchronous (including shortened QRS duration) can also be benefited from CRT.

Objective:To provide microsurgical anatomy data in the course, branch, distribution, arterial network profile of the submental artery and the range of the flap excision in submental flap transplantation.Methods:From March, 2015 to March, 2020, a total of 36 head and neck cast specimens were studied. Acrylic-butadience-styrene plastic (ABS) filler were perfused into the external carotid artery to make cast specimens. The course, branching, distribution and the arterial framework of the submental artery under a surgical microscope were investigated.Results:The submental artery originated from the facial artery before reaching the lower edge of the mandible (1.50±0.50) cm, with a diameter of (1.50±0.85) (0.6-2.3) mm. The main trunk of submental artery was (5.5±0.5) cm in length, which ran forward along the lower edge of the mandible and branched out (9.0±3.0) (7-13) branches with diameters between 0.1-0.5 mm, and mainly distributed to skin and superficial fascia of the submental area. The main trunk of submental artery divided into ascending, horizontal and descending branches about 3.0 cm of the midline of the mandible. The ascending branch went upwards over the lower edge of the mandible and joined up with the lower labial arch or participated in the formation of the lower labial arch; the horizontal branch divided into several branches and joined up with the branches from the opposite side; the descending branch branched posteriorly and inferiorly, joined up with branches of lingual artery and superior thyroid artery. The branches of the submental artery and the branches of the peripheral arteries were joined up in the submental area to form the submental artery network. The diameter of the vessels in the network ranged 0.1-0.2 mm. The arterial network was built in the form of 1 to 3 layers, and the area of main network was about 7.0 cm×5.0 cm.Conclusion:The submental artery has a long trunk, many branches and abundant anastomoses between the branches, forming a dense submental artery network, which provides sufficient pedicle length, rich blood supply and cutting area for submental flap. The flap can be transplanted free or transposed. The best location of submental flap is near the midline of arterial network, and the appropriate area is 7.0 cm×5.0 cm.

Objective To study the diagnosis and treatment of Mycobacterium avium complex lung disease with severe renal insufficiency,and to provide a basis for improving the clinical diagnosis and treatment of MAC lung disease with severe renal insufficiency.Methods Clinical data of an elderly patient with MAC lung disease confirmed by induced sputum culture and complicated with chronic kidney disease (CKD 4) was reported.The related literatures at home and abroad were reviewed.Results A male patient aged 80 years with poor nutrition was suffering from progressive weight loss,exhaustion and night sweats.His weight was only 43-44 Kg.Chest CT showed that multiple small nodules,small spot pieces of shadow and ground glass shadows in bilateral lung tissues.PET/CT indicated that spot pieces and nodules with metabolic activity in high performance.Bronchoalveolar lavage fluid (BALF) inspection was negative.MAC was identified by induced sputum culture through high permeability brine induced sputum.It was difficult to choose drugs for the treatment of MAC lung disease due to his poor kidney function [GFR of left kidney:9.0 ml /min,GFR of right kidney:18.8 ml/min].Conventional anti-mycobacterium drugs showed a low to high resistance to MAC.Moxifloxacin was discontinued for renal insufficiency.His condition was stable after choosing trimodality therapy including azithromycin 0.25 g/d tiw,rifapentine 0.3 g/d biw,ethambutol 0.375 g /d tiw and the joining immunotherapy.Conclusions PET/ CT is not useful in identifying tuberculosis and MAC lung disease.The treatment of MAC lung disease is difficult in elderly patients with severe renal insufficiency and low weight.Individual therapy combined with immunotherapy and improving the nutrition state is a good choice for the treatment of MAC lung disease in elderly patients with severe renal insufficiency.The anti-MAC drugs should be carefully selected and the adverse reactions should be closely observed in order to obtain the satisfactory clinical results.

Objective To investigate the clinical phenotype and genotype in three probands with antithmmbin(AT)deficiency and their families,and to identify the molecular mechanism of AT deficiency.Methods Chromogenic substrate method and immunoturbidimetry assay was used to detect the plasma levels of AT:A and AT:Ag,respectively.Genomic DNA was extracted from the peripheral blood.All 7 exons and the flanking sequences were amplified by PCR.and the abnormal mutant genes were analyzed by direct sequencing.Western blot was used to detect the AT levels and thrombin generation tests were used to detect coagulation status.Results The plasma levels of AT:A and AT:Ag of the three probands declined by 50%.G7386C(Trp225Cys)mutation in exon 4,C2591G(Ser36stop)in exon 2 and C9819T(Arg359stop)in exon 5 were characterized in the three prebands and they could result in W(Trp)225C(Cys)missense mutation,S(Set)36X(stop)nonsense mutation and R(Arg)359X(stop)nonsense mutation respectively,The testing results of phenotype and genotype from some of their family members showed consistent with results from the probands.Western blot results indicated that the Icyels of PC:Ag were lower compared with the normal pooled plasma.The hypercoagulative status was present in the probands using thrombin generation tests.Conclusions Type Ⅰ hereditary AT deficiency was found in these three families.The 3 heterozygous mutations.W225C,S36X and R359X are genetic defects of hereditary AT deficiency.W225C and S36X have not been described before.