Objective:To evaluate dendritic cells induced immune response against hepatocellular carcinoma by pulsed CD34+ hematopoietic stem cells originated dendritic cells with p53 gene.Methods:CD34+ hematopoietic stem cells were harvested after mobilization by chemotherapy and G-CSF.CD34+ hematopoietic stem cell apheresis was induced to differentiate into dendritic cells by cytokine cocktail IL-4,GM-CSF and TNF-?.On day 7,dendritic cells were transfected with plasmid pEGFP-C3/p53 DNA.The CTL response triggered by p53 pulsed dendritic cells was assayed by MTT method.Results:Dendritic cells originated from CD34+ cell apheresis had typical dendritic stick and expressed high level CD1a,CD11c,CD80,CD86,and HLA-DR molecules.After being pulsed with p53 gene,dendritic cells expressed green fluorescence protein and immunofluorescence assay(Cy3 labeled anti-P53 antibody)showed that transfected dendritic cells emitted red fluorescence.Dendritic cells inducing CTL response against HMCC97 cells(P53 positive)and HepG2 cells(P53 negative)were assessed by MTT method.P53 pulsed dendritic cells could induce P53 specific immune response against HMCC97 cells and the cytotoxin rate was(49.3?4.6)% compared with pEGFP-C3 transfection group [(25.4?4.1)%] and control group[(24.8?3.8)%](P0.05).Conclusion:P53 gene transfecting hematopoietic stem cell apheresis originated dendritic cells could induce specific CTL response against P53-expressing hepatocellular carcinoma cells.P53 may be a potential candidate for dendritic cell based immunotherapy of cancer.

Objective:To observe the effect of conversion from immediate-release tacrolimus (Tac) to extended-release Tac on kidney function in stable kidney transplant recipients.Methods:83 stable kidney transplant recipients who were converted from immediate-release Tac to extended-release Tac in the second people's hospital of Shanxi province from December 2011 to June 2019 were followed up for 12-36 months, and 83 stable kidney transplant recipients who continued to take immediate-release Tac were selected as control group.The changes of kidney function indexes, Tac trough concentration intra-patient variability (IPV) and compliance, the incidence of rejection and the survival rate of grafts and recipients were observed after the conversion from immediate-release Tac to extended-release Tac in stable kidney transplant recipients.Results:The conversion time from immediate-release Tac to extended-release Tac was (42.76±30.50)months after transplantation. At 24 months after conversion, the serum creatinine (SCr) was significantly lower than that before conversion ( P=0.013), and the estimated glomerular filtration rate(eGFR)was significantly higher than that before conversion( P=0.005). In the experimental group , the SCr was significantly lower than that of the control group at 36 months after conversion ( P= 0.017), eGFR was significantly higher than that of the control group ( P=0.038). In the experimental group, the score of Immunosuppressant Therapy Barrier Scale (ITBS) was (20.23±2.89) before conversion and (17.63 ±3.08) after conversion ( P= 0.000). The daily dose of Tac was (2.09 ±0.84) mg before conversion and (2.10 ±0.83) mg after conversion. The trough concentration of Tac before conversion was (7.22 ±2.84) ng/mL, which reduced significantly after conversion. No rejection occurred after conversion, and the recipients/grafts survived healthily during the follow-up period. Conclusions:After conversion from immediate-release Tac to extended-release Tac in stable kidney transplant recipients, the kidney function is stable and better than that of before conversion, the compliance is significantly improved, the IPV of Tac trough concentration is significantly reduced, and long-term use of extended-release Tac has good clinical efficacy and safety.

OBJECTIVE:To optimize the purification technology of total alkaloid from the flos of Aconitum kusnezoffii. METH-ODS:The content of total alkaloid from the flos of A. kusnezoffii was determined by acid-base titration. The purification technology of total alkaloid from the flos of A. kusnezoffii was optimized by ion resin with resin type,mass concentration of loading liquid and exchange speed as factors,maximum adsorption quantity,desorption rate and mass fraction of total alkaloid as index,and verifica-tion test was conducted. RESULTS:The optimal purification technology was as follows as type 732 cation exchange resin,mass concentration of loading liquid 0.32 g/L,exchange speed of 7 column volume(BV)/h. In validation test,the content of total alka-loid was 86.88%(RSD=0.52%,n=3),and desorption rate was 92.81%(RSD=0.40%,n=3)averagely. The extraction trans-port rate of total alkaloid from 3 batches of the flos of A. kusnezoffii was 81.76% and purification transport rate was 89.47% in av-erage. CONCLUSIONS:The established method is stable and feasible,and shows high transport rate.

Objective To summarize the safety and efficacy of low dose of hepatitis B immunoglobulin (HBIG) for prevention of de novo hepatitis B infection after renal transplantation.MethodsThe clinical data of 138 patients who received renal transplantation without hepatitis B infection between January 2007 and June 2010 were retrospectively studied (study group).All the patients in study group were given low dose of HBIG injection before transplantation.The HBsAb titer was monitored regularly after transplantation,and the dosage of HBIG adjusted according to the level of the HBsAb titer.HBIG was implied to all patients in the study group for more than one year.The clinical data of 196 patients who received renal transplantation without hepatitis B infection between January 2004 and December 2006 served as the control group.These 196 patients were not treated with HBIG.The incidence of de novo hepatitis B infection,and acute rejection of these two groups was analyzed.The one-year graft and patients survival rate was also investigated.Results During the follow- up period of 12 months,only one case in the study group had de novo hepatitis B infection (0.7％) 6 months after renal transplantation,while 11 cases (5.6％) in the control group had de novo hepatitis B infection,in which 2 cases were died from acute hepatic failure.The incidence of de novo hepatitis B infection had statistically difference between the two groups (P＜0.05).The incidence of acute rejection in the study and control groups was 13.8％ and 17.3％ respectively (P＞0.05).The one-year graft and patient survival rate in the study and control groups was 96.4％ and 97.8％,and 90.3％ and 91.8％ respectively (P＜0.05).ConclusionLow dose of HBIG is effective and safe for prevention of de novo hepatitis B infection after renal transplantation.

Ultrasonic elastography is a new technique of ultrasonic diagnosis and plays an important role in the diagnosis and monitoring of chronic liver diseases in recent years. This article introduces the principle and classification of ultrasonic elastography, as well as the application value of different techniques of ultrasonic elastography in liver diseases and related research advances. It is pointed out that ultrasonic elastography can analyze the stiffness of liver tissue qualitatively and quantitatively, which makes up for the shortage of traditional ultrasonic imaging and enriches diagnostic information. Therefore, ultrasonic elastography has an important clinical value and holds promise for wide clinical application.

Objective To explore the changes of mineral and bone metabolism before and after renal transplantation as well as the effect of preoperative parathyroid hormone (PTH) level on postoperative mineral and bone metabolism.Methods In this retrospective analysis,we recruited 82 cases of renal transplant recipients with normal renal function and receiving kidney transplantation in our hospital from January 2011 to January 2015.All of these patients had intact PTH (iPTH) level ＞300 pg/mL.We chose 26 cases of recipients whose preoperative iPTH was more than or equal to 800 pg/mL as very high PTH group,and 56 cases of recipients whose preoperative iPTH was between 301-799 pg/mL as high PTH group.We monitored and performed analysis of the total serum calcium (Ca),serum inorganic phosphorus (P),25-(hydroxyl) vitamin D3 (25 OHD),serum alkaline phosphatase (ALP),Beta C-terminal telopeptide (β-CTX),N-terminal/midregion (N-MID) pre-and 1 month,4 months,1 year,2 years,3 years post-kidney transplantation.Results Serum total calcium in the two groups was gradually increased,returned to normal range 1 month post-transplantation and reached the plateau 4 months post-transplantation.The incidence of hypercalcemia in very high PTH group was statistically significantly higher than in high PTH group.Serum phosphorus in the two groups showed a trend of gradual decline after renal transplantation,and returned to the normal range 1 month post-transplantation.The serum phosphorus level in very high PTH group reached the plateau 4 months post-transplantation,and that in high PTH group 1 month post-transplantation.Compared with high PTH group,very high PTH group has greater The incidence of long-term hypophosphatemia after renal transplantation was significantly higher in very high PTH group then in high PTH group.iPTH,ALP,β-CTX and N-MID in the two groups showed a downward trend after renal transplantation.At first month post-transplantation,iPTH,ALP,β-CTX and N-MID levels were reduced most significantly.The average levels of the three mentioned indicators in very high PTH group were higher than in high PTH group at every time point after surgery with the difference being significant during the early post-transplantation period.The anomalies of iPTH and β-CTX levels persisted to long term after transplantation in very high PTH group.25-OHD levels in these two groups showed rising trend after renal transplantation,reached the plateau 4 months posttransplantation,but failed to achieve the ideal reference level,and no significant difference was found between two groups at any time point monitored.Conclusion The anomalies of mineral and bone metabolism after renal transplantation could persist a long time.Conclusion hyperparathyroidism in the renal transplantation plays an important role in mineral and bone metabolism.Preoperative severe HPT could continue to post-transplantation period and increase the incidence of hyperphosphatemia and hypocalcemia long term after transplantation,which may aggravate bone turnover and this effect can last a long time after transplantation.

Objective To analyze the molecular epidemiology of adenovirus ( ADV) causing acute respiratory diseases and to investigate the mixed infection of ADV and other respiratory pathogens in Hebei Province. Methods Sputum samples were collected from inpatient children with acute respiratory diseases at Children′s Hospital of Hebei Province between June 2017 and May 2018. Multiplex reverse transcription PCR assay was used to detecte 13 kinds of respiratory pathogens. Nested PCR was performed to amplify ADV hexon gene and the amplified products were then sequenced. Results A total of 353 ADV-positive speci-mens were detected in 8839 specimens with a positive rate of 3. 99％. Significant difference in the positive rate of ADV was not observed between male and female patients (χ2=0. 0003, P=0. 99), but found among different age groups (χ2=115. 69, P<0. 001). All isolated ADV strains belonged to 11 serotypes, which were type 1 (16. 15％, 57/353), type 2 (35. 98％, 127/353), type 3 (21. 25％, 75/353), type 4 (1. 13％, 4/353), type 5 (11. 33％, 40/353), type 6 (3. 97％, 14/353), type 7 (8. 22％, 29/353), type 31 (0. 28％, 1/353), type 41 (0. 28％, 1/353), type 55 (0. 28％, 1/353) and type 57 (1. 13％, 4/353). Among the 353 ADV-positive specimens, 259 were mixed infections mainly caused by ADV and human rhinovirus (35. 52％). ADV and respiratory syncytial virus co-infections accounted for 12. 74％ and 33. 20％ of the mixed infections involved three or more pathogens. ADV could be detected throughout the year, especially in September and April to May. The predominant serotypes were types 1, 2 and 3. The av-erage ages of the two groups of ADV infection alone and ADV mixed infection were (27. 56±24. 67) months and (21. 33 ±20. 28) months, respectively, and the difference between them was statistically significant (P=0. 037). The incidence of ADV 2 infection alone was 25. 77％ (25/97), which was lower than that of ADV 2-involved mixed infection [39. 84％ (102/256),χ2=6. 05, P=0. 014]. However, the rate of ADV 7 infection alone was significantly higher than that of ADV 7-involved mixed infection [16. 49％ 16/97) vs 5. 08％ (13/256),χ2=6. 05, P<0. 001]. Conclusion ADV 1, ADV 2 and ADV 3 were the predominant serotypes circulating in Hebei Province from June 2017 to May 2018, especially in September and April to May. The younger the patients were, the higher the incidence would be. ADV 2 was prone to cause mixed infections with other respiratory pathogens, while ADV 7 was less common in mixed infections. Younger pa-tients were more susceptible to mixed infections. The most common co-infection was caused by ADV and hu-man rhinovirus.

Objective To explore risk factors of influencing operating efficiency of endoscopic submucosal dissection (ESD) for gastric mucosal lesions. Methods The data of 304 cases with gastric mucosal lesion undergoing ESD in Xijing Hospital of Digestive Disease from April 2009 to February 2017 were retrospectively analyzed. The procedure time and complete resection rate ( R0 resection rate ) were regarded as indicators of ESD efficiency. Risk factors influencing procedure time and R0 resection rate were analyzed using Chi-square test and logistic regression. Results Using median procedure time of 45 min as the cutoff value, Chi-square test showed that specimen size ( P=0. 000) , lesion location ( P=0. 001) , and pathological type ( P=0. 003) affected the operation time. Further logistic regression analysis indicated that specimen size (≥40 mm/<40 mm, P<0. 001, OR=3. 748, 95％CI: 2. 247-6. 254) and lesion location (upper or middle 1/3 of stomach/lower 1/3 of stomach, P=0. 001, OR=2. 180, 95％CI: 1. 318-3. 606) were independent risk factors of procedure time. Using R0 resection as outcome measure, neither single nor multiple parameter analysis was statistically significant. Conclusion Specimen size and lesion location are independent risk factors influencing efficiency of gastric mucosal ESD, and could be possibly used to estimate the procedure time of ESD.

Objective To access the influence factors of diagnostic delay of endometriosis. Methods We designed a questionnaire of diagnostic delay of endometriosis. From February 2014 to February 2016,400 patients who had dysmenorrhea and diagnosed with endometriosis by surgery in Peking University Third Hospital were surveyed retrospectively. Time and risk factors of diagnostic delay were analyzed.Results The diagnostic delay of 400 patients was 13.0 years(0.2-43.0 years),78.5%(314/400) patients thought pain was a normal phenomenon and didn′ t see the doctor. Patients who suffered dysmenorrhea at menarche experienced longer diagnostic delay than those who had dysmenorrhea after menarche(18.0 vs 4.5 years;Z=191.800,P<0.01).Patients who suffered aggravating dysmenorrhea experienced shorter delay time than those who suffered stable or relieving dysmenorrhea(11.0 vs 12.5 vs 18.0 years;Z=8.270, P<0.05), with the difference statistically significant, single factor analysis shows. Severe dysmenorrhea, deep infiltration endometriosis(DIE), family history of dysmenorrhea or endometriosis, previous surgical history of endometriosis,high stage,with infertility,adenomyoma or other symptoms,could help to shorten diagnostic delay with no significant difference(P>0.05). By multiple logistic regression analysis,the results shown that whether have dysmenorrhea at menarche and clinical diagnosis time were the independent factors affecting delayed diagnosis(P<0.01).Conclusions Diagnostic delay of endometriosis is common and the mean delay time is 13.0 years mainly due to the unawareness of dysmenorrhea. Dysmenorrhea at menarche,clinical diagnosis time and dysmenorrhea intensity are the factors affecting time of diagnostic delay.

Objective:To analyze the epidemiological characteristics of respiratory syncytial virus (RSV)-caused respiratory diseases in children in Hebei Province and the changes in peripheral blood lymphocyte subsets and inflammatory indexes.Methods:A total of 9 491 sputum specimens and 9 491 paired peripheral blood specimens were collected from children admitted to Hebei Children′s Hospital for respiratory infection in 2018. RSV-positive sputum specimens were screened by multiple detection reagents for 13 kinds of respiratory pathogens. Flow cytometry was uses to detect T and B lymphocytes and NK cells in peripheral blood samples of randomly screened RSV-positive children. Procalcitonin (PCT) was measured by Roche E411 luminescence analyzer. Hypersensitive C-reactive protein (hs-CRP) was detected by Roche Cobas 8000 C701 biochemical analyzer. White blood cells (WBC) were measured by Sysmex XN-BN3 hematometer.Results:The positive rate of RSV in children with respiratory diseases was 13.08% in Hebei Province in 2018. There were significant differences in RSV-positive rates among different age groups (χ 2=479.297 6, P<0.000 1). The positive rate of RSV decreased gradually with age (χ 2=-20.282 7, P<0.000 1) and was higher in male than in female (χ 2=34.552 7, P<0.000 1). The incidence of co-infection of RSV with other respiratory pathogens was 29.49% (366/1 241), mainly caused by human rhinovirus (HRV, 150/1 241) and adenovirus (ADV, 40/1 241). The main epidemic seasons of RSV infection were winter and spring. The epidemic trends of simple RSV infection and co-infection were consistent. There were significant differences in inflammatory indexes, WBC ( P<0.01), CD4 + ( P=0.015) and CD4 + /CD8 + cells ( P=0.016) between simple RSV infection and co-infection groups. Conclusions:RSV was a common pathogen causing respiratory diseases in children in Hebei Province. The younger the children were, the more likely they were to be infected with RSV. RSV infection was easily complicated by HRV or ADV infection. The epidemic seasons of RSV infection in Hebei were winter and spring. Both simple infection and co-infection of RSV might result in immune dysfunction.