Objective: To obtain chimeric antigen receptor macrophages ( CAR-M) targeting HER2 stably transfected． Methods : CAR lentivirus vector targeting HER2 was constructed and infected with human monocytic leukemia cell line (THP-1) ．CAR THP-1 cells with green fluorescent labeling were selected by sorting flow cytometry and continued to be cultured in vitro．The CAR THP-1 cells targeting HER2 were co-cultured with the endometrial cancer cell line Ishikawa with negative and positive HER2 expression，and their targeted phagocytosis of CAR-M to HER2 positive tumor cells was detected by imaging flow cytometry ，and the targeted phagocytosis efficiency of CAR-M to HER2 positive tumor cells was detected by flow cytometry. Results : CAR lentivirus infection with THP- 1 cells was less efficient ; After co-culture with cancer cells，flow cytometry and imaging flow cytometry showed that CAR THP-1 cells had enhanced phagocytosis of HER2 positive Ishikawa cells compared with the empty body group (P＜0. 01) ． Conclusion In this experiment，CAR THP-1 cell line targeting HER2 was established by constructing CAR lentivirus vector and transfecting THP-1 cells ，and it was proved that CAR THP-1 could phagocytize HER2 positive Ishikawa cells through specific targeting.

Objective:To evaluate the effectiveness of digital orthognathic surgery in the surgical treatment of patients with hemifacial microsomia (HFM) and patients’ satisfaction.Methods:The clinical data of HFM patients admitted to the Department of Orthognathic & Cleft Lip and Palate Plastic Surgery, Hospital of Stomatology, Wuhan University, from January 2017 to May 2020 were retrospectively analyzed. The digital orthognathic surgery was used to design surgical protocols before surgery. The intermediate wafer and terminal wafer by three-dimensional printing were applied to determine the position of maxillary and mandible bone blocks. The distance change between landmarks in maxillary and chin and the reference planes was measured. The skull CT and face photographs were taken 5 days after surgery to compare the distance between the actual position and the designed position. Data were expressed as the Mean±SD and analyzed by the pairing t-test with P<0.05 considered statistically. The patients’ satisfaction was investigated by interval scale method on day 7 and 6 months after operation. Results:There were 9 HFM patients in this study, included 5 men and 4 women, and the average age was 25.8±3.8 years old. 6 patients were affected on the left side and 3 patients was affected on the right side. All the operations were processed successfully. The wafers were in good position that the maxillary and mandible blocks were moved precisely according to the digital design. There were no accidental fractures of the jaw during the operation. The post-operative photographs and CT showed the stomatognathic system recovered well without serious postoperative complications. The errors between postoperative situation and preoperative designed situation in maxillary and chin were no statistically difference ( P>0.05). The maximum movement error of the maxillary bone block was the mark point of the first molar on the left, with an average error of (0.92 ± 0.34) mm. The patients’ satisfaction scores were averaged 83.2±2.7 points on day 7 after surgery, while the score dropped to 73.8±2.5 points after 6 months. Conclusions:The digital orthognathic surgery technology can satisfy the accuracy requirement of the surgical plan design for HFM patients in correcting the deflect of dental middle line, occlusal plane and chin point. High postoperative satisfaction can be achieved, which may decrease slightly 6 months after operation.

Objective : The live cell application was used to observe the process of phagocytosis of endometrial cancer cells by macrophages, and flow cytometry was used to detect the effects of macrophages engulfing tumor cells on activation of cytotoxic T cell. Methods : Ishikawa cells and THP1-induced macrophages were labeled with CFSE fluorescent probe and CD11 b respectively, and then mixed and seeded on a glass imaging dish.The live cell application was performed to record the phagocytosis of Ishikawa cells by macrophages within 120 minutes.Flow cytometry was used to detect the effect of macrophages engulfing tumor cells on activation of cytotoxic T cell. Results : The green fluorescence of Ishikawa cells was taken up by macrophages after the co-cultured two types of cells were in contact with each other, and macrophages were able to engulf multiple Ishikawa cells continuously.Macrophages that engulfed Ishikawa cells could induce activation of cytotoxic T cell. Conclusion The live cell application was successfully conducted to construct an in vitro model of phagocytosis of tumor cells by macrophages, which provided a feasible experimental method for detecting the dual killing process of macrophages and T cells on tumor cells.

Objective:To evaluate the effectiveness of digital orthognathic surgery in the surgical treatment of patients with hemifacial microsomia (HFM) and patients’ satisfaction.Methods:The clinical data of HFM patients admitted to the Department of Orthognathic & Cleft Lip and Palate Plastic Surgery, Hospital of Stomatology, Wuhan University, from January 2017 to May 2020 were retrospectively analyzed. The digital orthognathic surgery was used to design surgical protocols before surgery. The intermediate wafer and terminal wafer by three-dimensional printing were applied to determine the position of maxillary and mandible bone blocks. The distance change between landmarks in maxillary and chin and the reference planes was measured. The skull CT and face photographs were taken 5 days after surgery to compare the distance between the actual position and the designed position. Data were expressed as the Mean±SD and analyzed by the pairing t-test with P<0.05 considered statistically. The patients’ satisfaction was investigated by interval scale method on day 7 and 6 months after operation. Results:There were 9 HFM patients in this study, included 5 men and 4 women, and the average age was 25.8±3.8 years old. 6 patients were affected on the left side and 3 patients was affected on the right side. All the operations were processed successfully. The wafers were in good position that the maxillary and mandible blocks were moved precisely according to the digital design. There were no accidental fractures of the jaw during the operation. The post-operative photographs and CT showed the stomatognathic system recovered well without serious postoperative complications. The errors between postoperative situation and preoperative designed situation in maxillary and chin were no statistically difference ( P>0.05). The maximum movement error of the maxillary bone block was the mark point of the first molar on the left, with an average error of (0.92 ± 0.34) mm. The patients’ satisfaction scores were averaged 83.2±2.7 points on day 7 after surgery, while the score dropped to 73.8±2.5 points after 6 months. Conclusions:The digital orthognathic surgery technology can satisfy the accuracy requirement of the surgical plan design for HFM patients in correcting the deflect of dental middle line, occlusal plane and chin point. High postoperative satisfaction can be achieved, which may decrease slightly 6 months after operation.

Objective: To establish a method for isolation and culture of primary endothelial cells from non-human primate coronary arteries, and to provide a cell model for the study of human coronary endothelial cells. Methods: The coronary arteries of macaca mulattas were separated aseptically. The primary endothelial cells were separatedviatissue adhesion after collagenase digestion. CD31 positive cells were detected and sorted by flow cytometry to determine the purity of endothelial cells. After stimulation with prostaglandin E2(PGE2), the cellular viability and proliferation ability of primary coronary endothelial cells from macaca mulattas were evaluated by high-content cell imaging and CCK-8 assay, and the migration ability and tube function of primary coronary endothelial cells from macaca mulattas were measured by Transwell method and Matrigel glue method, respectively. Results: The confluence percentage of primary coronary artery cells of macaca mulattas was about 80% after 10-14 daysin vitroculture, and the cellular morphology was irregular polygons and paver shape. The purity of endothelial cells was about 31.7% by flow cytometry. After sorting, the purity of endothelial cells was confirmed by flow cytometry, which was more than 95%. PGE2could significantly up-regulate the proliferation, migration and tube formation abilities of primary coronary endothelial cells of macaca mulattas. Conclusion This study successfully established the isolation and culture method of primary coronary endothelial cells from macaca mulattas, and proved that it could be used as anin vitrocell model to simulate human coronary endothelial cells through functional studies.

Objective: To compare the efficiency of different methods for extracting rhesus monkey lung fibroblasts and their effects on functions, so as to provide a method for obtaining primary lung fibroblasts that are closer to human fibroblasts. Methods: Two extraction methods for rhesus monkey lung fibroblasts were used, direct tissue block adhesion method and collagenase combined digestion with tissue block adhesion method. The cell morphology was observed with the inverted microscope, the purity of isolated rhesus monkey lung fibroblasts was identified by immunofluorescence, cell viability was detected by CCK-8, the expression of α-SMA was detected by flow cytometry and the effect of long-term in vitro culture on cell apoptosis was detected by apoptosis kit. Western blot was used to detect the expression of α-SMA protein. Results: The combined digestion with collagenase and tissue block adhesion method could see small and bright cells crawling out in 24 hours, and cells could be seen crawling out in a large area after 48 hours. The cells were in a long spindle shape, after 4 days to 5 days, a single layer of cells could be formed. Identified by immunofluorescence, all cells expressed α-SMA. Tissue adhesion method showed small and bright cells crawling out after 72 hours. After 4 days to 5 days, the cells crawled out in a small area and showed a long spindle shape. After a week, the cells crawled out in a large area and formed a single layer of cells and the cells are all expressed α-SMA by immunofluorescence. The experimental results showed that the cell viability of the cells crawled out by the collagenase digestion method was significantly higher than that of the tissue adhesion method. After TGF-β1 stimulates the cells, the cells extracted by collagenase digestion method proliferated faster and expressed α-SMA more obviously. Conclusion Both methods can isolate rhesus monkey lung fibroblasts in vitro, but the collagenase digestion method extracts cells in a shorter time and in better condition. The expression of related proteins is more stable after stimulation by stimulants, which is an effective method to obtain rhesus monkey lung fibroblasts, and it is also an effective method to obtain primary lung fibroblasts that are closer to human.

OBJECTIVETo systematically evaluate the efficacy and safety of electroacupuncture (EA) for post stroke cognitive impairment (PSCI).

METHODSThe randomized clinical trials (RCTs) regarding EA for PSCI published before October of 2016 were researched in China National Knowledge Infrastructure (CNKI), Chinese Biomedical Database (CBM), WanFang database, VIP medicine information system, PubMed and Cochrane Library. The literature screening and information extraction was conducted by two independent reviewers. The quality assessment was performed based on the guidance of the Cochrane Reviewers' Handbook, and Meta-analyses was performed by using RevMan 5.3 software.

RESULTSTotally 14 RCTs were included, involving 896 PSCI patients. The results of Meta-analyses showed the EA group was superior to the control group in improving the MMSE [=1.78, 95%(0.24, 3.32),=0.02], the MoCA [=1.92, 95%(0.96, 2.88),<0.000 1], P300 latency [=-11.01, 95%(-18.91, -3.11),=0.000 6], P300 amplitude [=1.56, 95%(1.14, 1.98),<0.000 01], FMA score [=10.74, 95%(2.67, 18.82),=0.009] and the clinical effective rate [=1.37, 95%(0.98, 1.91),=0.06]. However, the comparison of BI score in both group had no significant differences [=6.38, 95%(-2.41, 15.18),=0.15].

CONCLUSIONThis Meta-analyses confirmed EA is effective and safe for PSCI, which could improve cognitive function and motor function. However, because of low quality of the included studies, more well-designed multicenter RCTs are needed.

Objective To investigate the effect of medicinal charcoal tablets combined with blood purification therapy on on renal function, inflammatory mediators, oxygenation index and intestinal barrier function in patients postoperative severe abdominal infection.Methods 65 cases with severe abdominal infection from the hospital were selected and randomly divided into control group (32 cases) and experiment group (33 cases) by random digital table method.The control group were treated by clinical routine therapy and experiment group were treated on the basis of control group with medicinal charcoal tablets combined with blood purification therapy.The renal function, inflammatory mediators, oxygenation index, intestinal barrier function and efficacy were tested.ResuIts Compared with control group after treatment, the renal function of serum creatinine (Scr), blood urea nitrogen (BUM), uric acid (UA) levels were lower (P<0.05), the inflammatory mediators of tumor necrosis factor α(TNF-α), interleukin-6 (IL-6) and interleukin-8 (IL-8) levels were lower (P<0.05), the oxygenation index (OI) was higher(P<0.05), the intestinal barrier function indicators of plasma diamine oxidase (DAO) and endotoxin of lipopolysaccharide (LPS) were lower (P<0.05), the total efficiency were higher (χ2 =3.91, P<0.05) in experimental group.ConcIusion The medicinal charcoal tablets combined with blood purification could effectively improve renal function, reduce inflammatory mediators levels, improve intestinal barrier function, and increase oxygenation index in patients with severe abdominal infection, which has a good clinical curative effect for postoperative severe abdominal infection .

Objective To observe the effect of electric acupuncture on the infarct volume and amount of cerebral cortex and spinal neuron at different times of cerebral ischemia/reperfusion(I/R)in stroke prone renovascular hypertensive rats(RHRSP)with middle cerebral artery occlusion(MCAO),and investigate the possible mechanisms of electric acupuncture on remote damage in ischemic stroke. Methods 480 male SPF Sprague-Dawley(SD)rats were duplicated to form the RHRSP models by clamping both kidneys. 370 successful ones were selected by taking the tail artery blood pressure,and divided into hypertension group and sham operation group(each n=60)by random number table method. The MCAO models were created by stringing middle cerebral artery in the remaining RHRSP. The nerve function defect score(NDS)was graded by Longa 5 point method after the rats waked up from anesthesia,then the ones scored 1-3 were enrolled. Totally,there were 190 rats with MCAO successfully created from which 10 were randomly selected to determine the infarct size by 2,3,5-triphenyl four azole nitrogen chloride(TTC)staining. The remaining 180 MCAO rats were randomly divided into model group,electric acupuncture group and fake acupuncture group(each n=60). The sham operated group only received surgical trauma;the electrical acupunctures atBaihuiandDazhuiacupoints on Du channel were performed on the day of model establishment in electric acupuncture group,once a day for 28 days;in fake acupuncture group,sticked the acupuncture needles at the skin ofBaihuiandDazhuipoints,then gived the same electrical acupuncture treatment. On 1,7,14 and 28 days after treatment,the rats of each group were respectively sacrificed,and the brains were collected,then the infarct volume and spinal neuron number were calculated by Nissl staining. Results ①Cerebral infarction volume:No infarcts were found in hypertension group and sham operated group. On 1 day and 7 days after MCAO,the infarct volumes were increased gradually in model group,electric acupuncture group and fake acupuncture group〔infarct volumes on 1 day were(12.36±0.11)%, (12.19±0.15)%,(12.24±0.16)%,and on 7 days were(20.01±0.24)%,(19.54±0.61)%and(19.77±0.25)%, respectively〕,and on 14 days and 28 days after MCAO,the infarct volumes were decreased gradually〔infarct volumes on 14 days were(17.18±0.23)%,(16.96±0.11)%,(17.08±0.62)%,and on 28 days were(14.38±0.21)%, (13.95±0.46)% and(14.29±0.37)%,respectively〕,but the differences among three groups had no statistical significance(all P>0.05). ② Cortex area:On 1 day after MCAO,the differences in the cortical neuron numbers among all groups were not statistically significant(all P>0.05). On 7,14,28 days after MCAO,the cortical neuron number of sham operated group was more than that of hypertension group,but the difference had no statistical significance(all P>0.05). Compared with sham operated group,the cortical neuron number in model group began to increase significantly after 7 days;compared with model group,the cortical neuron number in electric acupuncture group was increased obviously(cell/HP,7 days:75.48±2.41 vs. 68.78±1.42,14 days:61.32±2.60 vs. 48.78±1.41,28 days:53.65±1.46 vs. 28.78±1.21,all P<0.05),while the cortex neuron number of fake acupuncture group was markedly reduced(7 days:67.75±1.43,14 days:47.50±1.25,28 days:27.50±1.25), but the differences had no statistical significance(all P>0.05).③Spinal cord area:On 1,7,14 days after MCAO, the differences of the spinal cord neuron numbers among all groups were not statistically significant(all P>0.05). On 28 days,compared with hypertension group,the cord neuron number of sham operated group was increased,but the difference had no statistical significance(P>0.05). Compared with model group,the cord neuron number in electric acupuncture and fake acupuncture groups was inecreased(cell/HP:21.32±1.60,16.17±1.05 vs. 15.02±1.18),the difference being statistical significant in electric acupuncture group(P<0.05)but no statistical significance in fake acupuncture group(P>0.05). Conclusions Generally,the secondary spinal(cervical part)neuron death occurs after cerebral infarction in rats. The therapeutic action of electric acupuncture may reduce the secondary spinal neuron damage at remote site after cerebral infarction,that is possibly the mechanism of electric acupuncture for the protection of brain in hypertensive rats from I/R injury.