BACKGROUND:B-lymphocytes are an important participant in the immunity system. Currently, magnetic beads and complement methods are mainly used to isolate and purify B-lymphocytes. However, these methods are costly or cause large cel damage and low purity, which need further improvement. OBJECTIVE: To explore the isolation and culture methods of B-lymphocytes from mouse spleen and to study suitable conditions for B-lymphocyte isolation and culture in vitro by using interleukin-4, lipopolysaccharide, CD3 monoclonal antibody or their combination. METHODS:B-lymphocytes from mouse spleen were isolated and randomly divided into seven groups, respectively treated with interleukin-4, CD3 monoclonal antibody, lipopolysaccharide, interleukin-4+CD3, interleukin-4+lipopolysaccharide, CD3+lipopolysaccharide, and no stimulation (control group). Flow cytometry was used to detect the changes in the number and proportion of T-lymphocytes, B lymphocytes, and their subpopulations under different culture conditions. RESULTS AND CONCLUSION:The number of lymphocytes peaked at 3-5 days after addition of interleukin-4. In the lipopolysaccharide group, the number of lymphocytes began to increase at 3 days, and then peaked at 5 days. T-lymphocytes disappeared after addition of CD3 monoclonal antibody, so relatively pure B-lymphocytes could be obtained after 2 days and the number of B-lymphocytes reached the peak at 3 days. The number of mature B-lymphocytes (B220+IgD+) increased significantly after addition of CD3 antibody. In al the conditions we tested, transitional B cel subset (B220+CD93+) disappeared completely after 24 hours of culture. Experimental results indicate that after addition of CD3 monoclonal antibody and interleukin-4, T-lymphocytes can be removed in mouse spleen cels cultured, but mature B-lymphocytes remain to survive and proliferate.

Objective To explore the feasibility of inter-laboratory comparison and trueness evaluation among clinical laboratories, and assess the quality of participants′measurement, by measuring the activity of alanine aminotransferase ( ALT ) in patient serum samples.Methods Method comparison study was used.Five patients serum samples, whose target values were assigned by two international candidate reference laboratory with reference method of ALT without pyridoxal phosphate, were measured by 23 routine laboratories.The bias between measurement result of each participant and the mean of reference laboratories was calculated, and then compared to allowable bias 6%.Calculate the mean value and the relative bias.Results Compared with the mean of reference laboratories, the maximum absolute value of bias among the 23 routine laboratories was 31.27%.The rate range which bias was less than the allowable bias was 26.09%-73.91 %.The bias acceptability of 8 participants were more than or equal to 80%;15 participants were less than or equal to 60%; and 3 participants were 0%.Conclusions Using patient serum samples and values assigned by reference method is an effective way to carry out inter-laboratory comparison and trueness evaluation.It can reflect the quality of measurement more truly.

Objective To evaluate the short-term efficacy and safety of sevelamer hydrochloride in treating maintenance hemodialysis (MHD) patients with hyperphosphemia.Methods A multicenter,open-labeled,self-control study was performed.Phosphate binders were discontinued during a two-week washout period.Patients with more than 1.78 mmol/L serum phosphorus after two-week washout period were eligible for the trial.The dose was adjusted every two weeks as necessary to achieve serum phosphorus control. Sevelamer hydrochloride was administered to 138 MHD patients for 10 weeks and a second two-week washout period followed.Results A total of 111 from 138 patients fulfilled the whole 14-week study. Mean serum phosphorus and calcium-phosphate products starte to decline after two-week sevelamer hydrochloride treatment. By the end of 10-week sevelamer hydrochloride treatment, mean serum level of phosphorus [(1.85±0.50) vs (2.57±0.54) mmol/L,P＜0.01],calcium-phosphate product [(4.16± 1.72) vs (5.79 ± 1.50) mmol2/L2,P＜0.01 ] and low density lipoprotein [(1.64±0.76) vs (2.31 ±0.87) mmol/L,P＜0.01] were significantly decreased,while the adjusted serum level of calcium and serum intact parathyroid hormone kept steady.Both serum phosphorus and calcium-phosphrus product increased after the second washout period, but the levels were still lower as compared to pre-treatment [(2.26±0.71) vs (2.57±0.54) mmol/L; (5.12±1.63) vs (5.79±1.50) mmol2/L2,P＜0.01].Of the 138 patients involved,214 episodes in 106 patients and 121 episodes in 89 patients were reported as adverse events and adverse drug reaction respectively. Gastrointestinal symptoms,of which most were mild or moderate,happened to 68.1％ (94/138) patients. Conclusions Sevelamer hydrochloride can control serum phosphorus and reduce the levels of calcium-phosphorus product and cholesterol.Slight gastrointestinal symptoms like constipation are common during the treatment.

Objective This study aims to investigate the sex difference in the hippocampus and parahippocampus in patients with bipolar disorder. Methods We acquired T1－weighted structural MRI from 133 bipolar type I patients (60 males) and 144 normal controls (81 males). The General Linear Model was used to examine the relationship between sex and brain volumes of the hippocampus and parahippocampus, with age and intracranial volume as covariates. Results Patients showed significantly smaller volumes of the bilateral hippocampus and parahippocampus (P＜0.01). There were sex－by－diagnosis interactions in the left parahippocampus gyrus (F=6.534, P=0.044). Male patients had significant smaller volumes of the left parahippocampus gyrus compared to the male normal controls (P＜0.001) whereas the volumes were not significantly different between female patients and female normal controls (P＞0.05). Conclusion The results suggest sex difference in the left parahippocampus gyrus volume in patients with bipolar type I disorder, which deserves further investigation in the future bipolar imaging researches.

OBJECTIVEThis study aims to investigate the relationship between the dental and basal arch forms of patients with skeletal class II malocclusion using three-dimensional virtual models and to generate the best-fit curve for these arch forms.

METHODSThe mandibular dental casts of 35 skeletal class II patients were taken prior to treatment and were laser scanned. The facial axis (FA) points, which were used to represent the dental arch, and the WALA points, which were used to represent the basal bone, were identified for each tooth from the right first molar to the left first molar. The Pearson correlation coefficients between the intercanine and intermolar widths at the FA and WALA points were determined. Finally, the best-fit curves were generated using the fourth-degree polynomial equation to represent the arch forms.

RESULTSA strong correlation was found between the FA and WALA intermolar widths (r = 0.873). However, a moderate correlation was observed between the FA and WALA intercanine widths (r = 0.534). The difference between the intermolar widths at the FA and WALA points was significant (P < 0.01). No statistically significant difference was found between the intercanine widths (P = 0.481). The radii of curvature of the FA and WALA curves in the anterior teeth area were 15.194 and 20.250, respectively. The regression coefficients of the FA and WALA curves were calculated as R2 = 0.912 and 0.947, respectively.

CONCLUSIONFor skeletal class II malocclusion patients, the FA and WALA dental intermolar widths showed strong correlation, whereas the FA and WALA intercanine widths exhibited moderate correlation. The FA points were located more labially in the anterior teeth area but more lingually in the posterior teeth area. No statistically significant difference was found between the intercanine FA and WALA widths. The difference between the intermolar widths at the FA and WALA points was significant. The radii of curvature of the FA curve was larger than that of the WALA curve in the anterior teeth area.

Adolescent ; Cephalometry ; Dental Arch ; Female ; Humans ; Lasers ; Male ; Malocclusion ; Mandible ; Molar ; Tooth

Objective: To investigate the clinical effect of modified three minimally invasive blepharoplasty of double-fixation method. Methods: From April 2015 to December 2016, 82 patients were included, all received minimally invasive blepharoplasty. Three 3 mm-long small incision (at medial, middle and lateral) were designed, then part of the orbicularis oculi muscle and pretarsal fat were removed. Pretarsal fascia was then sutured with orbicularis oculi muscle and skin with two-layer-suture to form double eyelid respectively. Results: With up to 18 months follow-up, all patients had short period of detumescence within 7-12 days, high satisfaction rate with long lasting and natural double eyelid shape were achieved, no obvious scarring was noted. Conclusions Modified three minimally invasive blepharoplasty of double-fixation method has presented the merits of accurate presentation, natural shape, quicker recovery and less trauma, which makes it worthy of clinical presentation.

Objective:To explore the selection criteria of the donor for islet transplantation of Chinese people by analyzing the correlation between pancreas characteristics and success rate of islets isolation.Methods:Data from 113 cases of human islet isolation were collected. According to the result of islet isolation, the donors were divided into two groups, the success group(IEQ≥250 000, purification≥30%, and viability≥80%), and the failure group(IEQ<250 000, or purification<30%, or viability<80%). The modified Ricordi method was used to digest pancreas tissue, and the continuous density gradient method was performed to purify islets. The islets were identified by staining with the Dithizone(DTZ), the islets were analyzed for cell viability and purity.Results:The donor age in success group was significantly younger than failure group in the range of age eligible for this study( t=2.479, P=0.015). Pearson correlation showed that donor age was positively corelated with islet yield( r=-0.214, P=0.047). There was more fat on the pancreas surface in the successful islet isolation group( z=-2.007, P=0.045). The digestibility( t=2.133, P=0.035) and recovery rate( t=5.912, P=0.001) were elevated in success group. Conclusion:The pancreases from younger donors could obtain the higher-yielding islet, the pancreas with more surface fat or with higher weight was associated with islet isolation success in the scope covered by the inclusion criteria of this study.

Objective This study aims to investigate the relationship between the dental and basal arch forms of patients with skeletal class Ⅱ malocclusion using three-dimensional virtual models and to generate the best-fit curve for these arch forms. Methods The mandibular dental casts of 35 skeletal class Ⅱ patients were taken prior to treatment and were laser scanned. The facial axis (FA) points, which were used to represent the dental arch, and the WALA points, which were used to represent the basal bone, were identified for each tooth from the right first molar to the left first molar. The Pearson corre-lation coefficients between the intercanine and intermolar widths at the FA and WALA points were determined. Finally, the best-fit curves were generated using the fourth-degree polynomial equation to represent the arch forms. Results A strong correlation was found between the FA and WALA intermolar widths (r=0.873). However, a moderate correlation was observed between the FA and WALA intercanine widths (r=0.534). The difference between the intermolar widths at the FA and WALA points was significant (P<0.01). No statistically significant difference was found between the intercanine widths (P=0.481). The radii of curvature of the FA and WALA curves in the anterior teeth area were 15.194 and 20.250, respectively. The re-gression coefficients of the FA and WALA curves were calculated as R2=0.912 and 0.947, respectively. Conclusion For skeletal class Ⅱ malocclusion patients, the FA and WALA dental intermolar widths showed strong correlation, whereas the FA and WALA intercanine widths exhibited moderate correlation. The FA points were located more labially in the anterior teeth area but more lingually in the posterior teeth area. No statistically significant difference was found between the intercanine FA and WALA widths. The difference between the intermolar widths at the FA and WALA points was significant. The radii of curvature of the FA curve was larger than that of the WALA curve in the anterior teeth area.

Objective To retrospectively compare the efficacy of Serva NB1 collagenase with Vitacyte GOLD collagenase on islet isolation of pancreas.Methods All the human pancreata were obtained from Chinese organ donors.In GMP laboratory,the pancreata were trimmed and distended with Serva NB1 collagenase (Serva NB1,n =12) or Vitacyte GOLD collagenase (Vitacyte GOLD,n =5) and digested according to a modified Ricordi semi-automatic protocol,and the digestion duration was recorded.The digested islets were then collected and washed,followed by the continuous density purification in a Cobe 2991 cell separator.The islet yield,purity,viability and glucose-stimulated insulin release (GSI) were determined each time after purification.Quantity and quality of isolated islets were determined by digestion efficacy.Results The digestion duration in Vitacyte GOLD collagenase group was significantly shorter than in Serva NB1 collagenase group to achieve the same digestion endpoint (P＜ 0.05).The islets yields of different sizes were variable between the two groups.The Vitacyte GOLD collagenase digestion produced more islets with a diameter range of 50-100 μm than the ServaNB1 collagenase digestion (P＜0.05),but the latter yielded more islets with a diameter range of 251-300 μm and 301-350μm (P＜0.05).There was no significant difference in total islets yields,viability,and GSI between two collagenase digestions (P＞0.05).Conclusion Both Vitacyte GOLD collagenase and Serva NB1 collagenase can be used for the clinical islet isolation in China.

Objective:To investigate the correlations of β cell dedifferentiation in non-diabetic subjects with risk factors for type 2 diabetes mellitus(T2DM).Methods:Immunofluorescence staining with insulin and β cell dedifferentiated marker ALDH1A3 was used to evaluate the β cell dedifferentiation levels in 38 non-diabetic and 23 T2DM. Correlation analyses were performed between β cell dedifferentiation levels and available clinical parameters including age, body mass index, HbA 1C level, triglycerides, and cholesterol levels in non-diabetic subjects. Results:β cell dedifferentiation level defined by the positive expression of ALDH1A3 in β cells(ALDH1A3 + INS + cell proportion) was significantly elevated in T2DM subjects( P<0.001). In PreD subjects, ALDH1A3 + INS + cells proportion were decreased( P=0.050) and negatively correlated with HbA 1C( r=-0.44, P=0.006), but not with age and body mass index. The analysis of correlation with lipidemic parameters showed that ALDH1A3 + INS + cells proportion was positively correlated with plasma total cholesterol level( r=0.39, P=0.045), but not plasma total triglyceride. Conclusion:ALDH1A3 + INS + cells were found to be decreased in prediabetes, suggesting that there may be enhanced β-cell identity in prediabetes to compensate for insulin secretion requirements; ALDH1A3 + INS + cells were elevated in people with high plasma total cholesterol levels, suggesting that total cholesterol may be one of the factors that induce β-cell dedifferentiation.