Objective To investigate the differences and the immunocompatibility of wild-type (WT), four-gene modified (TKO/hCD55) and six-gene modified (TKO/hCD55/hCD46/hTBM) pig erythrocytes with human serum. Methods The blood samples were collected from 20 volunteers with different blood groups. WT, TKO/hCD55, TKO/hCD55/hCD46/hTBM pig erythrocytes, ABO-compatible (ABO-C) and ABO-incompatible (ABO-I) human erythrocytes were exposed to human serum of different blood groups, respectively. The blood agglutination and antigen-antibody binding levels (IgG, IgM) and complement-dependent cytotoxicity were detected. The immunocompatibility of two types of genetically modified pig erythrocytes with human serum was evaluated. Results No significant blood agglutination was observed in the ABO-C group. The blood agglutination levels in the WT and ABO-I groups were higher than those in the TKO/hCD55 and TKO/hCD55/hCD46/hTBM groups (all P<0.001). The level of erythrocyte lysis in the WT group was higher than those in the ABO-C, TKO/hCD55 and TKO/hCD55/hCD46/hTBM groups. The level of erythrocyte lysis in the ABO-I group was higher than those in the TKO/hCD55 and TKO/hCD55/hCD46/hTBM groups (both P<0.01). The pig erythrocyte binding level with IgM and IgG in the TKO/hCD55 group was lower than those in the WT and ABO-I groups. The pig erythrocyte binding level with IgG and IgM in the TKO/hCD55/hCD46/hTBM group was lower than that in the WT group and pig erythrocyte binding level with IgG was lower than that in the ABO-I group (all P<0.05). Conclusions The immunocompatibility of genetically modified pig erythrocytes is better than that of wild-type pigs and close to that of ABO-C pigs. Humanized pig erythrocytes may be considered as a blood source when blood sources are extremely scarce.

Objective To observe the value of ultrasound-guided intra-articular injection of hypertonic glucose(HG)for treating knee osteoarthritis in plateau area.Methods Ninety-one patients with knee osteoarthritis in plateau area(124 affected knee joints)were enrolled.The knee joints were divided into 20％HG group(n=67),25％HG group(n=42)or sodium hyaluronate group(n=15)based on the medication.Clinical and ultrasound scores were compared before and after treatment,and the efficacy of injection of HG was evaluated.Results At the 12th and 48th weeks after treatment,visual analog scale(VAS)scores decreased in all 3 groups(all P＜0.05),and Lysholm scores of 20％HG group and 25％HG group increased compared to those before treatment(all P＜0.05).The difference of Lysholm score before and in the 12th week after treatment,and of VAS score before and in the 48th week after treatment of 25％HG group were higher than those of 20％HG group(both P＜0.05).The joint exudation score of 20％HG group decreased in the 48th week after treatment compared to that before treatment(P＜0.05),and the synovial blood flow score decreased in the 12th and 48th weeks after treatment compared to those before treatment(both P＜0.05).The joint exudation score of 25％HG group decreased in the 48th week after treatment compared to that before treatment(P＜0.05),and the synovial hyperplasia and synovial blood flow score decreased in the 12th and 48th weeks after treatment compared to before treatment(all P＜0.05).The joint exudation score of sodium hyaluronate group decreased in the 48th week after treatment compared to that before treatment(P＜0.05),and the synovial hyperplasia score decreased in the 12th and 48th weeks after treatment compared to that before treatment(both P＜0.05).Conclusion Ultrasound-guided intra-articular injection of HG,esp.25％HG,had certain value for treating knee osteoarthritis in plateau area.

The aim of this study was to investigate the growth characteristics of primarily cultured astrocytes and microglia of different generations and then optimize the method for obtaining primary astrocytes and microglia effectively. Primarily cultured microglia were isolated and purified from the cortices of neonatal mice. The proliferation curve of mixed glia cells was measured by Cell Counting Kit-8 (CCK-8) assay, the proportion of astrocytes and microglia was detected by flow cytometry, and the polarization of the two types of glia cells was identified by immunofluorescence staining. Cell growth results showed that the mixed glia cells of P0 and P1 generation had the best proliferative activity; 97.3% of the high purity microglia could be obtained by mechanical shaking at 170 r/min for 30 min, and there was no significant difference in the morphology of ionized calcium-binding adapter molecule 1 (Iba-1) positive microglia and the proportion of M1 and M2 phenotype among the P0, P1 and P2 generations of microglia isolated by the above methods. Moreover, 95.7 % of the high purity astrocytes could be obtained by astrocyte cell surface antigen-2 (ACSA-2) magnetic beads separation, and there was no significant difference in the morphology of glial fibrillary acidic protein (GFAP) positive astrocyte and the proportion of A1 and A2 phenotype among the P0, P1 and P2 generations of astrocyte isolated by the above methods. Taken together, this study observed the growth characteristics of primarily cultured microglia and astrocyte in vitro, and then proved the best generations for purifying microglia and astrocytes. Finally, we optimized the methods of obtaining microglia and astrocyte, and verified that continuous culture within 2 generations will not affect the functional phenotypes of glia cells. These results provide technical support for studying the molecular mechanism of inflammation-associated diseases in nervous system.
Mice ; Animals ; Astrocytes/metabolism* ; Microglia/metabolism* ; Cell Count ; Flow Cytometry/methods* ; Cell Proliferation ; Cells, Cultured

Objective:To investigate the protective effect of hypothermic antegrade machine perfusion against canine ischemic brain injury.Methods:Thirteen beagle dogs were divided into the mild hypothermia with perfusion group ( n=6) and normothermia with perfusion group ( n=7) according to the random number table. The model of ischemic brain injury was established by neck transection. After 1 hour of ischemic circulatory arrest, the perfusion fluid based on autologous blood was continuously perfused through bilateral common carotid artery for 6 hours. The temperature of the perfusion fluid was set at 33 ℃ in the mild hypothermia with perfusion group and 37℃ in the normothermia with perfusion group, respectively. Blood oxygen saturation was recorded at 0, 1, 2, 3, 4, 5 and 6 hours after the beginning of perfusion to evaluate the perfusate oxygen level. The perfusate was collected, and the levels of Na +, K +, Ca 2+ and glucose as well as the pH value of the perfusate were detected in the two groups. At the end of perfusion, the parietal brain tissues of 1 dog from each group were collected to evaluate the water contents of brain tissues. Nissl staining was used to evaluate the morphological integrity of the pyramidal neurons in the frontal cortex and hippocampus. Neuronal nuclei antigen (NeuN) was used to evaluate the structural and morphological integrity of pyramidal neurons. Immunofluorescence glial fibrillary acidic protein (GFAP) and ionic calcium binding adaptor molecule 1 (Iba1) were used to evaluate the integrity and activity of astrocytes and microglia fragments. Results:At 0, 1, 2, 3, 4, 5 and 6 hours of perfusion, there was no significant difference in the blood oxygen saturation or Na + concentrations between the two groups (all P>0.05); the K + concentrations in the mild hypothermia with perfusion group were (4.57±0.12)mmol/L, (4.67±0.14)mmol/L, (4.27±0.12)mmol/L, (4.45±0.10)mmol/L, (6.60±0.15)mmol/L, (7.37±0.18)mmol/L and (9.03±0.16)mmol/L, respectively, which were significantly lower than those in the normothermia with perfusion group [(4.84±0.10)mmol/L, (5.31±0.13)mmol/L, (5.44±0.24)mmol/L, (5.70±0.18)mmol/L, (7.79±0.18)mmol/L, (10.44±0.40)mmol/L, (10.40±0.41)mmol/L] (all P<0.01). At 0, 1, 2 and 3 hours of perfusion, the Ca 2+ concentrations in the mild hypothermia with perfusion group were (0.72±0.15)mmol/L, (1.55±0.16)mmol/L, (1.62±0.15)mmol/L and (1.88±0.15)mmol/L, respectively, being significantly higher than those in the normothermia with perfusion group [(0.41±0.13)mmol/L, (0.99±0.12)mmol/L, (1.29±0.13)mmol/L, (1.57±0.11)mmol/L] (all P<0.01), and no significant differences were found at other time points (all P>0.05). At 0, 1 and 2 hours of perfusion, the glucose concentrations in the mild hypothermia with perfusion group were (5.75±0.19)mmol/L, (5.17±0.15)mmol/L and (4.72±0.15)mmol/L, respectively, being significantly higher than those in the normothermia with perfusion group [(5.30±0.22)mmol/L, (4.89±0.20)mmol/L, (4.30±0.17)mmol/L] (all P<0.01), with no significant differences found at other time points (all P>0.05). At 2, 3, 4, 5 and 6 hours of perfusion, the pH values of the mild hypothermia with perfusion group were 7.32±0.06, 7.25±0.02, 7.23±0.02, 7.24±0.02 and 7.24±0.02, respectively, being significantly higher than those in the normothermia with perfusion group (7.26±0.01, 7.21±0.01, 7.17±0.02, 7.15±0.02, 7.08±0.02) ( P<0.05 or 0.01), with no significant differences at other time points (all P>0.05). The water content of brain tissues in the mild hypothermia with perfusion group was (74.9±0.4)%, which was significantly lower than (79.9±0.9)% in the normothermia with perfusion group ( P<0.01). Nissl staining showed that the pyramidal neurons in prefrontal cortex and dentate gyrus had good integrity in the mild hypothermia with perfusion group. NeuN immunofluorescence staining showed that the morphology and structure of pyramidal neuron cells in the mild hypothermia with perfusion group were better with clearly visible axons than those in the normothermia with perfusion group, whereas the cytosol was full and swollen with scarce axons in the normothermia with perfusion group. GFAP and Iba1 immunofluorescence staining showed that more structurally intact glial cells, more abnormally active cells, thickener axons and better axon integrity in all directions were found in the mild hypothermia with perfusion group than those in the normothermia with perfusion group. Conclusion:Compared with normal temperature antegrade mechanical perfusion, the mild hypothermia antegrade mechanical perfusion can protect canine brain tissue and alleviate ischemic brain injury by maintaining stable energy and oxygen supply, balancing ion homeostasis and perfusion fluid pH value, reducing tissue edema, and maintaining low metabolism of pyramidal neurons, astrocytes and microglia.

Objective:To examine whether multiple-course prophylactic intravesical instillation chemotherapy is superior to single-course chemotherapy in preventing bladder tumor recurrence after radical nephroureterectomy(RNU)for primary upper tract urothelial carcinoma(UTUC).Methods:A comprehensive literature search was performed using the PubMed, Wanfang Data and China National Knowledge Infrastructure and all publications before March 2021 about clinical trials comparing the effects of single-course and multiple-course prophylactic intravesical instillation chemotherapy after RNU for UTUC were retrieved.Analysis was performed using Stata/SE 12.0.Results:Seven trials included a total of 998 patients, with 473 participants receiving multiple-course and 525 receiving single-course intravesical instillation chemotherapy.Bladder cancer recurrence occurred in 69 out of 473 patients(14.6%)with multiple-course chemotherapy and in 123 out of 525 patients(23.4%)with single-course chemotherapy.The absolute risk reduction was 8.8% and the relative risk reduction was 37.6%.Compared with a single-course instillation, the pooled OR of bladder cancer recurrence was 0.55(95% CI: 0.40-0.76, P<0.001)for multiple-course instillations.No serious adverse events were reported. Conclusions:Compared with single-course instillation chemotherapy, multiple-course prophylactic intravesical instillation chemotherapy significantly decreases the risk of bladder cancer recurrence in primary UTUC patients treated with RNU, indicating multiple intravesical instillation chemotherapy is superior to single instillation chemotherapy.

Objective To explore the application effect of exercise rehabilitation based on the Health Participation Model in patients with lung cancer surgery.Methods A total of 108 patients with lung cancer hospitalised in our hospital between January and December 2022 were randomly divided into two groups:a control group and a intervention group,with 54 patients per group.The control group was offered with routine exercise rehabilitation,while patients in the research group underwent exercise rehabilitation based on the Health Participation Model.Results One month after discharge,the research group showed higher levels in the first second maximum expiratory volume(FEV1),forced vital capacity(FVC),maximum ventilatory volume(MVV)and FEV1/FVC ratio in comparison with those in the control group(all P<0.001).The total score of postoperative rehabilitation quality and the scores of its dimensions were all higher in the research group than those in the control group one month after discharge(all P<0.001),except the dimension of psychological support.Conclusion Incorporation of the Health Participation Model into the postoperative rehabilitation program for lung cancer patients can improve lung function and promote the postoperative rehabilitation.

Objective:To understand the research progresses of economic evaluation of non-pharmaceutical interventions (NPIs) both at home and abroad, and provide reference for economic evaluation of NPIs using real-world data in China.Methods:The literature retrieval was conducted by searching Chinese and English databases to indude papers about economic evaluation of NPIs and integrated NPIs published from January, 2020 to December, 2021, and the results were analyzed comprehensively.Results:A total of 30 Chinese and English literatures about economic evaluation of NPIs for COVID-19 prevention and control were included; including 7 papers about nucleic acid and testing and screening, 6 papers about individual prevention and protection measures, 12 papers about integrated implementation of individual prevention and protection, social distancing, nucleic acid or antigen testing, community screening and symptom screening, as well as close contact tracing and isolation/quarantine, and 5 papers about contain strategies, such as lockdown. This study found that personal protection, social distancing, and testing-tracing-isolation measures were cost-effective; however, different combinations of NPIs might lead to different results. Moreover, the cost of lockdown was high, which might cause huge economic burden.Conclusions:Most NPIs are cost-effective except lockdown, while the cost-effectiveness of the integrations of NPIs at different levels and in different scenarios needs to be further evaluated. It is necessary to carry out economic evaluation of integrated NPIs and the combination of NPIs with other interventions, such as vaccination and medication, based on real-world settings in China.

Objective To observe the efficacy and safety of recombinant human growth hormone ( rhGH) replacement therapy in GHD childhood with craniopharyngioma after surgery. Methods This study retrospectively reviewed the records of 18 inpatients with secondary GHD diagnosed by insulin tolerance test ( ITT ) after craniopharyngioma surgery at the Department of Endocrinology, Beijing Tiantan Hospital, from January 2012 to December 2015. The clinical benefits and risks of 18 patients were evaluated systematically, and then were divided into rhGH treatment group(n=9) and control group(n=9). The parameters of height, growth velocity(GV), height standard deviation score (HtSDS), insulin-like growth factors-1 (IGF-Ⅰ), insulin-like growth factor binding protein 3 (IGFBP3) and adverse events rate were recorded after treatment for six months. MRI was followed up every 3 to 6 months to observe the difference of the tumor recurrence and second malignant neoplasm between two groups. Results All 18 patients with craniopharyngioma presented with multiple pituitary-target glands hormone deficiency after surgery. Among these patients, 17 cases (95% ) presented with hypothyroidism or adrenal insufficiency, 7 cases (39% ) with delayed puberty, and 12 cases(67% ) with central diabetes insipidus. Based on pituitary-target gland axis function deficiency, these patients were given appropriate L-thyroxine, prednisone, and desmopressin(DDAVP) replacement therapy, respectively. The median time of 9 patients starting rhGH replacement was 48(36,72)months after surgery. The levels of height, GV, IGF-Ⅰ, HtSDS, and IGFBP3 were significant increased after rhGH treatment for 6 months as compared with pre-treatment and control group (all P<0. 05). In addition, changes of height, GV, HtSDS, and IGF-Ⅰ levels before and after treatment in rhGH treatment group were significantly higher than those in control group (all P< 0. 05). During the period of treatment, there were no serious adverse events to be observed in rhGH treatment group, except 1 case of mild headache in the control group. The differences of biochemical and endocrine parameters such as thyroid function, liver and renal function, blood glucose, etc. were without statistical significance compared with pre-treatment after rhGH treatment ( P > 0. 05 ). The tumor recurrence and second malignant neoplasm were not detected by MRI scanning in rhGH treatment group,but there were 3 cases in the control group. Conclusion Multiple pituitary-target glands axis deficiencies were observed in childhood patients with craniopharyngioma after neurosurgery, and the evident deficiency of GH-IGF-Ⅰ axis was observed. rhGH replacement therapy in short-term would significantly improve the parameters of growth and development of patients with GHD after craniopharyngioma neurosurgery. No recurrence tumor in situ and second malignant neoplasms were detected during the period of rhGH replacement therapy.

Objective To explore the impact of silver ions on the adhesion ability of Bacillus Calmette-Gueri vaccine on silicone tube surface. Methods Common and silver plated silicone tubes were cultured in BCG bacterial liquid for 30 days. A gas-bacilli was taken once every 3 days to measure OD value of microbial. The growth curve of BCG vaccine was made. The corresponding silicone tubes in divided different generations was removed and eluted based on the division cycle of BCG vaccine. The samples were inoculated on the solid medium for observing the colony growth amount, and adhesion curve was drawn. The silicone tube colony line was observed under the electron microscope at day 30. Results OD value and bacterial plaque number were less in silver silicone tube group than in common silicone tube group, with statistical significances (P < 0.05). Electron microscopy also showed that BCG adhesion on silver silicone tube group was relatively less than that on common silicone tube group. Conclusion Silver ions can reduce the adhesion of BCG vaccine on silicone tube surface and inhibit the growth of BCG vaccine.

Objective To explore the functional mechanism of a Chinese medicine compound “Qiangzhizufang”on rat model of Tourette syndrome ( TS) combined with fear.Methods The rat model of TS combined with fear was established by intraperitoneal injection of 3,3’-iminodipropionitrile (IDPN) combined with acoustic stimulation.After giving different drug lavage treatment, the changes of behavior of the rat models were assessed by field test and behavior test.The content of DA, TH and TH mRNA in the brain tissue was detected by HPLC, immunohistochemistry and RT-PCR, separately.Results Compared with the normal control group, stereotyped behavior and exercise behavior were increased, freezing time prolonged, but the content of DA, TH and TH mRNA in the brain tissue were not obviously changed in the model control group.Compared with the model control group, the stereotyped behavior and exercise behavior were decreased, content of DA, TH and TH mRNA in the brain tissue was decreased in the “Qiangzhizufang” group. Conclusions The Chinese medicine compound“Qiangzhizufang” can improve the behavior in rat models of TS combined with fear.This effect may be realized through down-regurating TH mRNA expression, reducing the content of TH, and reducing the dopamine synthesis.