Objective·To investigate the regulatory effects and underlying mechanisms of mouse mandibular osteoblasts on B cell differentiation and development.Methods·Single-cell suspensions from mouse mandibular bone were prepared using an optimized enzymatic digestion method and induced to differentiate into osteoblasts in vitro.Osteogenic potential was validated by real-time quantitative PCR(RT-qPCR),alkaline phosphatase(ALP)staining,and alizarin red S(ARS)staining.The spatial localization relationship between osteoblasts and B cells in mandibular tissues was examined via immunofluorescence staining.High-purity hematopoietic progenitor cells were isolated using fluorescence-activated cell sorting.A Transwell co-culture system was established to assess the regulatory effects of different osteoblast concentrations(5×104,2.5×105,and 5×105 cells/well)on B cell differentiation(5×104 cells/well).Flow cytometry and RT-qPCR were employed to evaluate B cell viability and differentiation.Additionally,RT-qPCR was used to analyze the expression of osteoblast-secreted factors associated with B cell development during osteogenic differentiation.Results·Mandibular osteoblasts exhibited robust osteogenic potential,as confirmed by ALP/ARS staining and high expression of osteogenic markers(Runx2,Osx,Ocn,and Alp)via RT-qPCR.Immunofluorescence revealed close spatial proximity between osteoblasts and B cells in mandibular tissues.In the co-culture system,osteoblasts promoted B cell differentiation in a concentration-dependent manner.RT-qPCR and immunofluorescence demonstrated that osteoblasts significantly upregulated key genes involved in B cell development(Ebf1,Rag1,Il7r,and Pax5;all P<0.001).Furthermore,osteoblast-derived factors(Il7,Baff,and Flt3l)were markedly elevated during osteogenic differentiation(all P<0.05).Conclusion·Mandibular osteoblasts enhance B cell differentiation and development in a concentration-dependent manner,likely through secreting growth factors that upregulate critical B cell differentiation genes.

Objective·To investigate the regulatory effects and underlying mechanisms of mouse mandibular osteoblasts on B cell differentiation and development.Methods·Single-cell suspensions from mouse mandibular bone were prepared using an optimized enzymatic digestion method and induced to differentiate into osteoblasts in vitro.Osteogenic potential was validated by real-time quantitative PCR(RT-qPCR),alkaline phosphatase(ALP)staining,and alizarin red S(ARS)staining.The spatial localization relationship between osteoblasts and B cells in mandibular tissues was examined via immunofluorescence staining.High-purity hematopoietic progenitor cells were isolated using fluorescence-activated cell sorting.A Transwell co-culture system was established to assess the regulatory effects of different osteoblast concentrations(5×104,2.5×105,and 5×105 cells/well)on B cell differentiation(5×104 cells/well).Flow cytometry and RT-qPCR were employed to evaluate B cell viability and differentiation.Additionally,RT-qPCR was used to analyze the expression of osteoblast-secreted factors associated with B cell development during osteogenic differentiation.Results·Mandibular osteoblasts exhibited robust osteogenic potential,as confirmed by ALP/ARS staining and high expression of osteogenic markers(Runx2,Osx,Ocn,and Alp)via RT-qPCR.Immunofluorescence revealed close spatial proximity between osteoblasts and B cells in mandibular tissues.In the co-culture system,osteoblasts promoted B cell differentiation in a concentration-dependent manner.RT-qPCR and immunofluorescence demonstrated that osteoblasts significantly upregulated key genes involved in B cell development(Ebf1,Rag1,Il7r,and Pax5;all P<0.001).Furthermore,osteoblast-derived factors(Il7,Baff,and Flt3l)were markedly elevated during osteogenic differentiation(all P<0.05).Conclusion·Mandibular osteoblasts enhance B cell differentiation and development in a concentration-dependent manner,likely through secreting growth factors that upregulate critical B cell differentiation genes.

【Objective】 To investigate the clinical characteristics and antibody distribution as well as evaluate the transfusion efficacy in unexpected antibody positive patients. 【Methods】 A total of 12 235 patients from January 1, 2022 to March 31, 2023 who hospitalized in our hospital and applied for blood transfusion were selected, and those with unexpected antibody were included. The clinical data, including gender, age, diagnosis, blood type, history of transfusion and pregnancy were collected for antibody distribution analysis. Patients who received transfusion were grouped according to the DAT results and the components of red blood cells transfused, and the Hb values of each group before and after transfusion were compared. 【Results】 Among12 235 patients, 118 were positive for antibody screening, with a prevalence of 0.96%. The antibodies from Rh system were the most common (27.43%, 48/175), followed by MNS system (8.57%, 15/175) and Lewis system (6.29%, 11/175), mainly anti-E (18.29%, 32/175), anti-M (8.00%, 14/175) and anti-Le^a (5.71%, 10/175). In addition, 62 transfused patients were divided into group A with suspended red blood cell transfusion and group B with washed red blood cell transfusion for positive DAT, and group C for negative DAT. Hb values (g/L) pre- and post-transfusion were 59.19±15.67 vs 77.52±15.09 in group A, 56.35±14.08 vs 74.44±15.63 in group B, 56.00±12.06 vs 75.00±4.73 in group C, respectively. The Hb values of post-transfusion for three groups were all higher than those of pre-transfusion (P<0.05). 【Conclusion】 Anti-E from Rh system is the most common antibody in patients with unexpected antibody. Appropriate red blood cells transfusion with Hb increases by an average of 6-7 g/L per 1 U of red blood cells indicating good transfusion efficacy. For positive DAT patients, transfusion of suspended red blood cell is feasible.

Objective·To compare the effects and complications of carbon dioxide laser laryngectomy and open partial laryngectomy in the treatment of laryngeal carcinoma.Methods·Clinical data of 101 patients with stage Ⅰ-Ⅲ laryngeal squamous cell carcinoma admitted to Renji Hospital,Shanghai Jiao Tong University School of Medicine from January 2013 to January 2018 were selected and divided into open surgery group and minimally invasive group according to different surgical methods.The open surgery group received open partial laryngectomy(57 cases),and the minimally invasive group received carbon dioxide laser laryngectomy(44 cases).Follow-up visits were made by telephone and outpatient visits,and the operation time,postoperative laryngeal mucosa edema time,length of stay,5-year survival rate,complication rate and recurrence rate of the two groups were compared in the different clinical stages(stage Ⅰ,stage Ⅱ,and stage Ⅲ)and tumor locations(glottic type and non-glottic type).Results·The operation time,postoperative laryngeal mucosa edema time and length of stay of the minimally invasive group were shorter than those of the open surgery group(P＜0.05).Comparison of the 5-year survival rates of patients with the same clinical stage and glottic type between the two groups showed no statistically significant difference(both P＞0.05);while comparison of the 5-year survival rates of patients with non-glottic type between the two groups(62.5％in the open surgery group vs 0 in the minimally invasive group)showed a statistically significant difference(P＜0.05).In the minimally invasive group,there were 3 cases of complications,including 1 case of fever and 2 cases of lung infection;in the open surgery group,there were 14 cases of complications,including 5 cases of fever,7 cases of lung infection and 2 cases of pharyngeal fistula.The complication rate of patients in the minimally invasive group(6.82％,3/44)was lower than that in the open surgery group(24.60％,14/57),and the difference was statistically significant(P＜0.05);the complication rate of patients with stage Ⅱ and glottic type in the minimally invasive group was significantly lower than that in the open surgery group(P＜0.05).Comparing the recurrence rate of patients in the two groups(10.53％in the open surgery group vs 2.27％in the minimally invasive group),the difference was not statistically significant(P＞0.05).Conclusion·Compared with open partial laryngectomy,carbon dioxide laser laryngectomy has faster postoperative recovery and lower complication rate in the treatment of early and middle laryngeal malignant tumors,so it is worth promoting.

Objective To analyze the clinical efficacy of different anti-tumor therapies for recurrence and metastasis after liver transplantation for primary liver cancer (liver cancer). Methods Clinical data of 145 recipients undergoing liver transplantation for liver cancer were retrospectively analyzed. The overall survival and recurrence and metastasis after liver transplantation for liver cancer were analyzed. The clinical efficacy of different anti-tumor therapies for recipients with recurrence and metastasis were compared. Results Sixty-five recipients (44.8%) developed recurrence and metastasis. The median recurrence time was 6 months. Among them, 1 case underwent secondary liver transplantation after recurrence and died of intestinal perforation. Twenty-four recipients (37%) received targeted drug therapy with a median tumor-bearing survival of 22 months. Eleven recipients (17%) received radiotherapy or chemotherapy with a median tumor-bearing survival of 11 months. Nine recipients (14%) received local treatment (surgical resection or radiofrequency ablation), and the median tumor-bearing survival was 8 months. Twenty recipients (31%) abandoned anti-tumor therapy, and the median tumor-bearing survival was 3 months. The tumor-bearing survival of recipients receiving anti-tumor therapy was significantly longer than that of recipients without anti-tumor therapy (P < 0.001). The tumor-bearing survival of recipients receiving targeted drug therapy was significantly longer than that of those receiving other anti-tumor therapies (P=0.03). The tumor-bearing survival of recipients receiving local treatment, radiotherapy and chemotherapy was considerably longer than that of those who abandoned anti-tumor therapy (P=0.004). Conclusions Surgical resection and radiofrequency ablation are the optimal therapies for recipients with recurrence and metastasis after liver transplantation for liver cancer. For recipients with multi-focal tumors who fail to receive local treatment, those receiving targeted drug therapy obtain the longest survival. In addition, radiotherapy and chemotherapy can also prolong the survival of recipients with recurrence and metastasis.

Objective To evaluate the uncertainty of the pseudo-ginsenoside GQ (PGQ) concentration in human plasma by HPLC-MS/MS.Methods The whole process of PGQ determination by HPLC-MS/MS in human plasma was evaluated and the uncertainty caused by repeatability,weighing,standard solution preparation,biological sample preparation,extraction recovery process,recovery,instrument precision and calibration curve fitting were evaluated,respectively.The combined and expanded uncertainty values were both calculated.Results The expanded uncertainty values for low (15.16 ng·mL-1),medium (2 516.67 ng·mL-1) and high (3 902.00 ng·mL-1) levels of PGQ were 1.39,177.74 and 262.69 ng·mL-1,respectively (P =95 ％,k =2).Conclusion The uncertainty of the PGQ determination in human plasma by HPLC-MS/MS is mainly caused by recovery,repeatabihty and sample preparation at low concentration,by sample preparation and recovery at medium and high concentration.

Objective To investigate the effects of Schisandrae Sphenantherae Fructus and processed with vinegar on lipid metabolism of rats with type 2 diabetes mellitus (T2DM). Methods The rat model of T2DM was induced by high fat diet plus STZ. The rats were randomly divided into blank group, model group, Schisandrae Sphenantherae Fructus high-dose group and low-dose group, and vinegar Schisandrae Sphenantherae Fructus high-dose group and low- dose group. The rats in each group were fed with the corresponding medicine for gavage for 30 d. FINS, FFA, TC, TG, HDL-C, LDL-C, and MDH, total protein content of liver tissue were detected. HE staining was used to observe the histomorphological changes of liver and pancreas in rats. Results Compared with the model group, Schisandrae Sphenantherae Fructus groups and vinegar Schisandrae Sphenantherae Fructus groups did not show obvious effects on improving FBG and FINS, but it could raise varying degrees of HDL-C and MDH, and reduce FFA, LDL-C, TC, and TG, among which vinegar Schisandrae Sphenantherae Fructus could significantly regulate metabolism in T2DM rats. Conclusion Vinegar Schisandrae Sphenantherae Fructus can enhance the lipid metabolism regulatory function of T2DM rats.

Objective To investigate the iodine nutritional status in the key populations before and after the adjustment of salt iodine content in Yantai of Shandong.Methods In 2010 (the pre-adjustment period) and 2014,2015 (the post-adjustment period),the changes in the residents' iodized salt,the goiter prevalence and urinary iodine of children aged 8-10,the urinary iodine of pregnant women,and the iodine content of drinking water before and after the adjustment were analyzed.Results The coverage rate of iodized salt and the edible rate of qualified iodized salt were 98.27％ and 97.28％,respectively before the adjustment of salt iodine content,and 97.44％ and 96.14％ after the adjustment.The mean of salt iodine after the adjustment (21.96 mg/kg) was significantly lower than that of 2010 (31.45 mg/kg,t =66.29,P ＜ 0.05).The goiter prevalence of children aged 8-10 by thyroid palpation was 0.92％ in 2010,while it was 1.89％ by ultrasonic in 2014,2015.There was significant difference in the iodine nutritional status of children in 2010 (191.0 μg/L) and in 2014,2015 (173.0 μg/L,Z =3.56,P ＜ 0.05).The difference of iodine nutritional status in pregnant women between pre-adjustment (154.0 μg/L) and post-adjustment (130.4 μg/L) was also significant (Z =5.54,P ＜ 0.05).The median of water iodine was 5.4 μg/L after the adjustment.There were 52 towns with medians of water iodine below 10 μg/L.Conclusions The coverage rate of iodized salt and the edible rate of qualified iodized salt have all met the national standard before and after the adjustment of salt iodine content.The mean of salt iodine during 2014,2015 is significantly lower than that of 2010.Before and after the adjustment,the goiter rates of children aged 8-10 are all below 5％.The adjustment of salt iodine content is more suitable to children aged 8-10 than to pregnant women currently.It is suggested that pregnant women eat more foods rich in iodine.

OBJECTIVE:To establish a method for determining recombinant human calmodulin B subunit(rhCNB)in rat plas-ma,and study its pharmacokinetics characteristics. METHODS:ELISA double-antibody sandwich method was adopted. 1 μg/ml rhCNB monoclonal antibody mAb was wrapped,added to the to-be-test sample,rhCNB polyclonal antibody pAb(dilution ratio of 1∶5 000)and HRP-labeled conjugate of anti-IgG(dilution ratio of 1∶10 000)were added. Using tetramethylbenzidine for develop-ing,microplate reader was conducted in wavelength of 450 nm to determine the absorbance value(OD value)and plasma concen-tration of 6 rats after 2,15,30,60,120,240,480,720 min of iv 2.5 mg/kg rhCNB,and the pharmacokinetic parameters were calculated by BAPP 3.0 software. RESULTS:The linear range of rhCNB were 0.195-12.5 ng/ml(r2=0.995 0),lower limit of quan-titation was 0.195 ng/ml,accuracy were 97.300%-103.622%(RSD<7.5%,n=6);RSDs of within-batch,inter-batch,freezing and thawing 3 times were no higher than 8.5%(n=6,18,15). rhCNB pharmacokinetics characteristics in rat fitted to two-com-partment model,AUC0-720 min was 173.038 mg·min/L and t1/2 was 94.62 min. CONCLUSIONS:The established method has high specificity and sensitivity,good accuracy and precision,which can be used for rhCNB quantitative detection and pharmacokinetics study in biological samples.

Objective:To investigate the diagnosis and treatment significance of invasive and noninvasive operation for ventilator?as?sociated pneumonia ( VAP ) . Methods:A total of 80 cases of VAP suspected patients who had received mechanical ventilation at least 48 hours in ICU from Jun 2014 to Mar 2015 were enrolled. Patients were randomly divided into four groups including noninvasive operation group ( F) , invasive operation group ( Q) , mix group 1 ( H1) and mix group 2 ( H2) . VAP diagnosis rate between groups as well as living time, antibiotic use time, survival rate, calcitonin levels and APACHE II score, oxygenation index were analyzed. Results:Specimen from invasive operation had higher specimens to cultivate positive rate than that from noninvasive operation ( P<0?05), but there was no statistic significant difference in VAP diagnosis rate between two methods (P>0?05). Conclusion:Noninva?sive operation collecting samples for VAP diagnosis is also accurate as invasive one. Collecting specimens from sputum suction tube in the clinical treatment on airway suction is a low cost and simple noninvasive operation.