BACKGROUND:At present,there are two main methods of isolating and purifying bone marrow mesenchymal stem cells(BMSCs):density gradient centrifugation and the whole bone marrow adherent method.The former has complicated procedures,and the latter has simple operation,but the purified outcomes are not ideal.OBJECTIVE:To establish a rat BMSCs isolated and cultured in vitro purification methods on the basis of the whole bone marrowadherence and isolation of BMSCs in combination of differential passage digestion method.METHODS:By whole bone marrow adherent culturing to isolate and differential digestion and passage of rat BMSCs,the speed ofMSCs in the process of digestion and passage was quicker than other bone marrow cells,as well as the characteristics of adherentspeed,instead of density gradient centrifugation to separate and purify MSCs,and their morphological characteristics wereobserved.Cell growth and proliferation of two kinds of culture method were compared with the density gradient centdfugationseparation.Alkaline phosphatase and oil red staining results were observed to verify BMSC differentiation capacity,to detect thecall surface markers,to validate immune properties and to test its purity.RESULTS AND CONCLUSION:The whole bone marrow adherent culture method isolated and differentially subcultured ratBMSCs.Flow cytometry and osteogenic and adipogenic culture results displayed cytoirnmunity property,purity and differentialcapacity,which have no significant difference compared with density gradient centrifugation.However,cell viability andreproductive activity are obviously elevated.

Objective To investigate the clinical effect of different nutritional therapies on the immune function of patients with malignant obstructive jaundice after percutaneous transhepatic cholangiodrainage (PTCD).Methods A total of 50 patients with malignant obstructive jaundice who were admitted to our hospital from January 2009 to March 2013 were randomly divided into two groups according to the admis-sion order.The patients in group A (n=25 )received enteral nutritional support after PTCD,and those in group B (n=25 )received total parenteral nutritional support after PTCD.Intra-group and inter-group comparisons were made in terms of jaundice clearance,nutritional indices,and body’s immune function on preoperative day 1 and postoperative day 7;comparison between the two groups was made by t test. Results Among the 50 patients who underwent PTCD,39 (78%)had good drainage,while 1 1 (22%)did not reach the expectation,of which,5 (10%)were in group A and 6 (12%)in group B.In both groups,the nutritional indices on postoperative day 7 were significantly higher than those on preoperative day 1(P<0.05),but no significant difference in these indices was observed between group A and group B on postoperative day 7 (P>0.05).The immune function of patients in both groups was significantly improved following PTCD and nutrition-al support (P<0.05),but there was no significant difference between the two groups (P>0.05).Although the same scheme of nutritional support was used,there were 1 1 patients who did not achieve the expected jaundice clearance after PTCD and had limited improvement in immune function compared with those who had complete jaundice clearance (all P<0.05).Conclusion Jaundice clearance is closely re-lated to PTCD in patients with malignant obstructive jaundice,but not markedly associated with the ways of nutritional support.

Objective To evaluate the method of hemodialysis patients with the diagnosis and treatment of herpes zoster.Methods Diagnosis and treatment of 27 cases of hemodialysis patients with varicella-zoster virus (VZV) infection in patients with clinical data.Results All patients in addition to continuing the line hemodialysis treatment,use the ganciclovir,topical recombinant human interferon α -1b cream,vitamin Bl,Methycobal,antibiotics to prevent bacterial infection,oral medicine,comprehensive treatment of pain.The total effective rate of treatment group is 92.59％.Conclusion VZV was a common disease in hemodialysis patients,easy diagnosis,rational drug treatment is very important,need comprehensive treatment can achieve good results.

Objective To explore effective treatment of uremic hemodialysis patients with chronic helicobacter pylori-negative erosive gastritis.Methods Fourty-eight cases of uremic hemodialysis patients with helicobacter pylori-negative chronic erosive gastritis were diagnosed by detection of helicobacter pylori and endoscopic examination.Fourty-eight patients were randomly divided into two groups,treatment group (24 patients) with oral pantoprazole 40 mg,once a day,combinding with oral Marzulene-S particles,each 0.67 g,three times a day,the control group (24 cases ) only with the dose of pantoprazole treatment.Effect of treatment was compared in these two groups after treatment for 21 days.Results Twenty-one days after treatment,the clinical symptoms of uremic hemodialysis patients with upper gastrointestinal improved in treatment group,and the performance of endoscopic and histopathological has varying degree of improvement.It shows that pantoprazole combined with Marzulene-S treatment group has a significant effect than that single treatment with pantoprazole group.In the treatment group,the total effective rate was 95.83％,however it was 83.33％ in the control group( U =2.716,P ＜ 0.01 ).Repair of mucosal lesions were significantly different between the two groups ( total effective rate 83.33％ vs 45.83％,U =2.349,P ＜ 0.05 ).Conclusion Pantoprazole combined with Marzulene-S therapy has a significant effect on uremic hemodialysis patients with chronic Helicobacter pylori-negative erosive gastritis.

Objective To evaluate the effect of interventional vas embolism operation at ICU bedside in severe pelvis fracture patients complicated with hemorrhagic shock. Method Forty-eight severe pelvis fracture patients with hemorrhagic shock were treated by interventional vas embolism operation at bedside as well as intensive monitoring. The clinical results were compared with those of the traditional conservative therapy group. Results In the interventional therapy group, 46 patients with hemorrhage had been controlled within an hour after the operation and the success ratio reached 95.8%. The blood transfusion volume, the complication incidence and mortality rate were all significantly lower than those of the conservative therapy group. Conclusion Interventional vas embolism operation at ICU bedside is a safe, practical and effective treatment on pelvis fracture with iliac vas trauma.

Objective To investigate the effects of high density lipoprotein (HDL) on thrombin-activated platelet al-pha-granulemembrane protein (CD62P) and lysosome intact membrane protein (CD63) expressions in vitro. Methods The equivalent volume of washed platelets prepared by hand was preincubated with HDL (1 g/L) in 37℃water for 15 minutes, which was then stimulated with different concentrations of thrombin (0.5 U/mL, 1 U/mL and 10 U/mL) for 10 minutes in wa-ter of 37℃. Meanwhile another three groups of washed platelets were incubated with thrombin (0.5 U/mL, 1 U/mL and 10 U/mL) for 10 minutes, respectively. The CD62P and CD63 from each sample were analyzed by flow cytometry (FCM). Results The CD62P positive rates of HDL-preincubated groups were significantly lower than those of different concentrations of thrombin groups (0.5 U/mL,1 U/mL and 10 U/mL) in the absence of HDL (11.55%± 1.34% vs 18.14%± 1.50%, 17.19%± 0.17% vs 26.24%± 0.77% and 19.79%± 0.32% vs 80.38%± 5.66%,P ＜ 0.01). Meanwhile, The CD63 positive rates of HDL-preincubated groups were also significantly lower than those of thrombin-treated (0.5 U/mL, 1 U/mL and 10 U/mL) groups without HDL, namely,2.92%±0.22%vs 8.09%±0.48%(P＜0.001), 4.20%±0.98%vs 14.15%±1.39%(P＜0.001) and 5.12%± 0.09% vs 24.48%± 1.71%(P ＜ 0.01). Conclusion HDL inhibits the expression of CD62P and CD63 on throm-bin-stimulated platelets in vitro.

Objective To observe the changes of the heart rate turbulence(HRT)in patients with isolated systolic hypertension(ISH).Methods 85 patients with ISH,80 patients with common hypertension and 58 control subjects were investigated.Turbulence onset(TO)and turbulence slope(TS)were recorded of 24 hours dynamic electrocardiogram.Results The difference of HRT in the three groups was significant(P＜0.01).The values of TO and TS was significantly higher in common hypertension group and ISH group than in control group(P＜0.05);TS was significantly lower in ISH group than in common hypertension group(P＜0.05),but the difference of TO between ISH group with common hypertension group was not obvious.Conclusion The HRT in patients with primary hypertension is significantly lower,which is more significant in patients with ISH than in patients with common hypertension.It suggests that the cardiac autonomic nerves dysfunction perhaps has an important influence on development of hypertension and its prognosis.

Objective:To discuss the clinical characteristics and genetic diagnosis of fetal familial hemophagocytic lymphohistiocytosis (FHL).Methods:Clinical data of a case of fetal FHL from Children's Hospital, Capital Institute of Pediatrics was analyzed, and related FHL cases at home and abroad were retrieved from PubMed, CNKI, and Wanfang databases using terms including "fetus", "neonate", and "familial hemophagocytic lymphohistiocytosis", from the establishment of the database to January 3, 2021, to summarize the characteristics of this disease.Results:This index case was found with fetal splenomegaly, free fluid in the abdominal cavity, and enlargement of the ventricle at 39 +3 weeks of gestation, and presented with fever, tachypnea, hepatosplenomegaly, skin ecchymosis and petechia, and lymphadenectasis after birth. Laboratory examination revealed pancytopenia, abnormal liver function, elevated ferritin and triglyceride, and decreased fibrinogen levels. CD107a excitation experiment showed decreased degranulation function of NK cell (ΔCD107a<5%). Hemophagocytosis was observed in the bone marrow smear. Genomic DNA sequence analysis demonstrated compound heterozygous mutations of c.118-308C>T and c.3002T>C in the UNC13D gene. All the above findings led to the diagnosis of FHL3. Despite chemotherapy with dexamethasone and cyclosporin, and symptomatic treatment after admission without hematopoietic stem cell transplantation, the baby died on day 52. A total of 15 papers related to fetal FHL, including 20 infants, were retrieved. Among these 21 cases (including the index case), the main clinical symptoms were fetal edema and hepatosplenomegaly, which may be accompanied by fetal distress and increased amniotic fluid volume, and postnatal fever, dyspnea, rash, and central nervous system involvement. Laboratory and imaging examination results were consistent with the diagnostic criteria for hemophagocytic hyperplasia. As far as we know, the reported fetal FHL gene mutations were PRF1 (FHL2) and UNC13D gene mutation (FHL3), in which reduced expression of perforin and granzyme can be detected, respectively. Dexamethasone, cyclosporin, etoposide, and other chemotherapy and symptomatic treatment are the primary treatments currently, and alternative therapies include intrauterine chemotherapy in the third trimester and postnatal hematopoietic stem cell transplantation. Among the 21 cases, including the index case, intrauterine death occurred in four cases, 13 children died at different times after birth, and only four children survived, among which the eldest one was 12 years old. Conclusions:FHL is a condition with atypical early signs, high mortality rate and treatment difficulties. Fetal FHL should be considered in differential diagnosis in fetuses with edema or hepatosplenomegaly besides hemolysis, infection, autoimmune diseases, and hereditary problems. Therefore, with immunotechnology and gene sequencing, early diagnosis and treatment can be prompted to improve the prognosis of this group of population.

【Objective】 To investigate the matrix effect on the determination of potency in Recombinant Human Coagulation Factor Ⅷ for Injection (rFⅧ). 【Methods】 Two different detection matrices were used to establish two methods for detecting the potency in Recombinant Human Coagulation Factor Ⅷ for Injection. And the matrix effect on the determination of potency was determined, including specificity, linearity, repeatability, accuracy and intermediate precision. 【Results】 As to the specificity, the recoveries of the two substrates at high vs low concentration level were 112% and 110% vs 104% and 109%, respectively. As to the linearity, in the range of (0.125-1.000) IU/mL, the correlation coefficient between concentration and coagulation time of standard/ sample was higher than 0.99. As to the accuracy/repeatability, the recoveries of two matrices was 104% and 102%, and RSD was 2.4% and 1.9%. As to the intermediate precision, personnel factor of two matrices was 0.72 and 0.23, date factor was 0.79 and 0.85, and RSD(for 12 times) was 4.2% and 3.0%. Comparison of two matrices was as follows: Deviation in test results of 6 batches of rFⅧ was all lower than 5%. There was no significant difference between two matrices. 【Conclusion】 The two matrices for potency detection show good performance including specificity, linearity, repeatability, accuracy, and intermediate precision. They are suitable for the determination of potency in rFⅧ products.