BACKGROUND:Bone marrow derived mesenchymal stem cells (BMMSCs) have multi-differentiation potentials, possessing a repairing capability for the sectional bone defects if combined with degradable porous β-tricalcium phosphate china. This provides a new idea in clinical repair of various bone defects.OBJECTIVE: To explore in radiology the curative effect of implanting porous β-tricalcium phosphate and autogenous BMMSCs compound to treat bone defects.DESIGN: A randomized controlled study with callus growth at various healing periods as subjects for observation. SETTING: Department of Orthopaedics, Renmin Hospital, Wuhan University; Tissue Engineering Center, Research Institute of Basic Medicine,Academy of Military Medical Sciences of Chinese PLAMATERIALS: This experiment was carried out at the Tissue Engineering Center, Research Institute of Basic Medicine, Academy of Military Medical Sciences of Chinese PLA between March and September 2002. Twenty China healthy adult sheep were randomized into the groups of blank control group (4 sheep), simple implantation group (8 sheep) and complex implantation group (8 sheep).METHODS: Under general anesthesia and aseptic condition, 10-15 mL of sheep marrow was extracted; MSCs were separated and cultured before combined with porous β-tricalcium phosphate china for tissue engineering bone construction. Rats in each group were cut off 21mm long metatarsus in the middle section of metatarsus bonestem. Β-tricalcium phosphate china and autogenous MSC compound was implanted into the sheep of the complex implantation group; β-tricalcium phosphate china was implanted into the simple implantation group; and the bone defects in the blank control group remained untouched. Then the incision was sutured.X-ray filming was carried out right after the operation, as well as 1, 3,and 6 months after the operation for radiological appraisal (scored for1 if bone union formed in one surface of bone defect, but scored 0 if no boneunion formed in any surface of bone defect, and scored 4 if bone union formed in front, back, lateral surfaces and the center of bone defect), Xray radiation-resisting density was analyzed to compare the results of bone defect repair.MAIN OUTCOME MEASURES: The postoperative general condition and general observation, as well as the results of the radiological analysis of the bone defects of all sheep.RESULTS: Totally 20 sheep were brought into this xperiment and all entered the stage of result analysis. ① The postoperative general condition:Sheep regained consciousness 2-6 hours after the operation without incision infection and loosing of internal fixation. Their spirit gradually was back to normal 1 week after the operation, at which time the injured legs could touch ground but were incapable of bearing load, and the affected legs could bear load 2 weeks after the operation, walked slightly lamely 3 weeks after the operation, and even moved freely without limp 4 weeks after the operation. ②The general condition 6 months after the operation: In pure implantation group, the surface white hyaline cartilage-like tissues were gradually calcified, with both ends connected with the host bone by bone bridge, but china granules could still be easily observed; while no implantation substance could be observed in compound implantation group,with the boundary between implantation substance and host bone vanished,and bone defect became basically the same as host bone. However there was no bone tissue formed in bone defect at various postoperative time points in the blank control group. ③ Radiological analysis of the bone defects at various postoperative time points: The radiological rating score was obviously higher in complex implantation group at the time poin ts of 3, 6 months after the operation compared with the pure implantation group [(2.3±0.3), (1.8±0.5); (3.3±0.5), (2.6±0.6), P ＜ 0.05]. ④ Radiological analysis of bone callus thickness and the relative value of radiation-resisting density at various postoperative time points: The bone callus thickness in the complex implantation group was obviously lower than that of the pure implantation group at the postoperative time point of 6 months (4.62 vs 7.64, P ＜ 0.05), with relative value of radiation-resisting density obviously higher than that of the pure implantation group (70.4±1.5 vs 61.18±1.2, P ＜ 0.05).CONCLUSION: Radiological appraisal and bone defect density measurement can well reflect the dynamical repairing process of bone defects; the implantation of porous β-tricalcium phosphate china and autogenous BMMSCs compound into sheep can enhance the repair of large sectional bone defect.

A new method using on-line cleanup technology combined with liquid chromatography tandem mass spectrometry ( UHPLC-MS/MS) was developed for the determination of seven kinds of β2-agonists residues, Formoterol, Salmeterol, Carbuterol, Clenisopenterol, Clenpenterol, Clencyclohexerol and Clenbuterol-hydroxymethyl in livestock manure. The sample was sufficiently extracted by acidic acetonitrile and diluted by 0. 2% formic acid. The extract was online purified on HyperSep Retain CX column where the sample matrix was washed away and the analytes were retained. The analytes were eluted into Hypersil Gold C18 column by 2% Ammonia-methanol solution. The seven β2-agonists were detected in selected reaction monitoring ( SRM) mode via positive electrospray ionization ( ESI+) by liquid chromatography-tandem mass spectrometry. The results showed good linearity correlation coefficients over 0 . 9928 for the seven kinds of β2-agonists in 0 . 5-100 μg/L concentration range. The LOD of seven kinds of β2-agonists in livestock mature is 1 μg/kg, while the LOQ is 5 μg/kg. When 5-50 μg/kg of the seven kinds ofβ2-agonists were added into the blank livestock manure, an average recovery of 67% -112% was obtained with the relative standard deviations of 2. 9%-10. 2%. The method is simple, rapid and has good reproducibility for quantitative and confirmatory analysis ofβ2-agonist residues.

To determine the residue of 14 sulfonamides in milk, a high performance liquid chromatography-tandem mass spectrometry ( HPLC-MS/MS ) method with on-line soild phase extraction ( SPE ) in cation exchange mode was established. 5 g of milk was extracted with 15 mL acetonitrile. Then the extraction was evaporated by 50 ℃ nitrogen and dissolved by 1. 00 mL 0. 2% formic acid. The dissolution was enriched and purified by MS/MS cation exchange on-line SPE column on a double ternary liquid chromatography, and eluted by the mixed solution of 2% ammonia methanol and 0. 2% formic acid (50:50, V/V). The compounds were separated by an octadecyl silica bonded column and determined by the tandem mass spectrometry. The results showed that the linearity of 14 sulfonamides was good in the range of 0 . 1–10 μg/kg ( r≥0 . 9995 ) .
The LOD of the method was 0. 05 μg/kg, while the LOQ was 0. 1 μg/kg. The recoveries of the 14 sulfonamides were in the range of 60 %-90 %, while the inter-batch and intra-batch RSDs were all lower than 10%. The method was proved to be more convenient, economical and stable than the traditional SPE column method.

Objective To explore curriculum and faculty qualification of Chongqing operating room (OR) specialty certified nurse theoretical tralning program by feedback of students, and to im-prove the quality of tralning. Method Relying on the national key project of the First Affiliated Hos-pital of Chongqing Medical University, Chongqing Health administration OR nurse specialist tralning program set additional courses related to research, teaching, patient safety and first ald tralning based on operation room specialist nurse tralning outline. 81 participants were given questionnalres issued by college of nursing after the 7th OR specialist nurse theoretical tralning, and statistics analysis was made of it. Result Effective questionnalres was100%. The overall judgment on theory tralning was high, with 92.60%of the students giving overall score above 150 points (out of 170);93.84%of students sat-isfied on teaching staff ratio while 95.06% students satisfied on course content. For the 17 items, the majority scores given by the students were ≥ 9 points (a 10-points scale), on the top of the list were item “Theoretical tralning helps to enhance professional theoretical level and guide clinical practice”“Teaching content meet needs of OR specialist nurses” “Teaching content is helpful to improve the core competence of specialist nurses” (all 96.30%, 78 out of 81), while the bottom was “Instructor create an environment which fosters student participation”(85.19%, 69 out of 81). Conclusion Chong-qing OR specialist theoretical tralning has good practical and guidance property, the content meet OR nurses needs, instructor ratio is reasonable, and students are highly satisfied with it. Instructors need to encourage students' participation in class interaction, and pay attention to improving their research capabilities.

ObjectiveTo evaluate the relationship between estrogen receptor-α-29 (ERα-29) gene polymorphisms and the development of HBV-related hepatocellular carcinoma (HCC) in Gansu Province, China, and to investigate the pathogenesis of HCC at the gene level. MethodsGene polymorphisms of ERα-29 were analyzed in 106 HBV-related HCC patients and 98 healthy individuals as normal controls using the polymerase chain reaction-restriction fragment length polymorphism technique. Population allele frequencies were calculated using the gene counting method and then tested using the Hardy-Weinberg law of genetic equilibrium. Comparisons of genotype and allele frequencies between groups were performed using the χ2 test. ResultsThe frequencies of TT genotype and T allele of ERα-29 gene in HBV-related HCC patients were significantly higher than those in the normal controls, i.e., 31.1% and 53.8% vs. 11.2% and 32.1% (χ2 = 3.449, P＜0.05; χ2 = 3.840, P＜0.05). In contrast, the frequencies of CC genotype and C allele of ERα-29 gene in HBV-related HCC patients were significantly lower than those in the normal controls, i.e., 23.6% and 46.2% vs. 47.0% and 67.9% (χ2 = 3.488, P＜0.05; χ2 = 3.840, P＜0.05). Compared with those carrying C allele, carriers of T allele had an increased risk (2.46-fold) of HBV-related HCC (OR = 2.46, 95% CI: 1.64-3.69). Conclusion T allele of ERα-29 gene can increase the risk of HBV-related HCC.

Objective To investigate the clinical features of hepatitis B core antibody (anti-HBc)positive patients with liver injury.Methods A total of 212 anti-HBc positive and HBsAg negative patients who were primarily diagnosed with liver injury from August 2013 to August 2014 at Ruijin Hospital were collected for this study.The patients were divided into cirrhosis group (n=60) and non-cirrhosis group (n =152) according to the status of cirrhosis.The 60 cases with cirrhosis were further compared with 60 cases with post-hepatitis B cirrhosis.The general information,biochemistry and immunology data were assessed.ANOVA was used to compare multiple groups of means,and Wilcoxon rank-sum test was used for non-parametric comparisons of the two groups.Results Only one case was positive for HBV DNA with the positivity rate of 0.5％.The causes for liver injury were as follows,60 cases with cryptogenic cirrhosis,which accounted for 28.3 ％;45 cases with drug-induced hepatitis,which accounted for 21.2 ％;33 cases with unexplained liver injury,which accounted for 15.6％;28 cases with acute hepatitis E,which accounted for 13.2％ and 15 cases with autoimmune hepatitis,which accounted for 7.1％.There were significant differences of T cell subpopulation,hepatitis B surface antibody (HBsAg) and hepatitis B e antibody (anti-HBe) quantitative level,red blood cells (RBC),platelet counts (PLT),prealbumin,albumin,alamine aminotransferase (ALT),aspartate transaminase (AST),international normalized ratio (INR),hyaluronic acid (HA),collagen Ⅲ (COL-Ⅲ) and collagen Ⅳ (COL-Ⅳ) between the cirrhosis group and non-cirrhosis group (all P＜0.05).The CD3+ CD4+ and CD3+ CD8+ counts,white blood cells (WBC),ALT,AST,total bilirubin (TBil) and albumin in anti-HBc-positive cirrhosis group were statistically different from those in post-hepatitis B cirrhosis group (all P＜0.05).Conclusions Some patients with positive anti-HBc still have HBV replication and infectivity.HBV anti-HBc positivity and HBsAg negativity may be associated with some cryptogenic cirrhosis and primary liver cancer.Patients with positive anti-HBc are prone to be complicated with drug-induced hepatitis,autoimmune hepatitis,and other liver damage related to immune mechanisms.Patients with cirrhosis have a higher risk to induce immune tolerance and progress to chronic disease than non-cirrhotic patients.Quantitative anti-HBc might be used as an indicator to predict disease progression after HBV infection.Disease condition in cirrhotic group with positive anti-HBc and negative HBsAg is less severe than that in post-hepatitis B cirrhosis group.

Objective:To explore the clinical efficacy and risk factors of umbilical cord mesenchymal stem cells (UCMSCs) infusion at an early stage (i.e.gross hematuria) for hemorrhagic cystitis (HC) after allogeneic hematopoietic stem cell transplantation (allo-HSCT).Methods:The relevant clinical data were retrospectively reviewed for 300 patients undergoing allo-HSCT from January 2016 to July 2021.According to the presence or absence of HC, they were assigned into two groups of HC (n=89) and non-HC (control, n=211). According to whether or not receiving an infusion of UCMSCs, 51 patients of HC degree Ⅱ-Ⅳ were divided into two groups of UCMSC infusion and non-infusion.The risk factors of HC after allo-HSCT were analyzed by χ2 test.Logistic regression was employed for multivariate analysis of P<0.05.Mann-Whitney U test was utilized for statistically analyzing the duration of gross hematuria and urinary tract irritation symptoms and evaluating the clinical efficacy of UCMSCs infusion for HC. Results:Among them, 89 (29.67%) developed HC post-allo-HSCT.Clinical grades were Ⅰ (n=38, 42.70%), Ⅱ (n=36, 40.45%), Ⅲ (n=13, 14.61%) and Ⅳ (n=2, 2.25%). The median occurrence time was 29 (21.5-35.0) days post-allo-HSCT.In univariate analysis, age ≤30 years, haploid transplantation, antithymocyte globulin (ATG), acute graft-versus-host disease (aGVHD), CMV-DNA positive pretreatment significantly boosted the risk of HC ( P<0.05). In multivariate analysis, aGVHD was an independent risk factor for HC ( OR=10.281, 95% CI: 1.606-65.813, P=0.014). Among 89 HC patients, 38 grade Ⅰ patients were complete remission(CR). Among 51 patients of grade Ⅱ-Ⅳ HC, the outcomes were CR (n=48) and non-remission(NR)(n=3). And 24/51 of them received UCMSCs plus conventional treatment.The duration of gross hematuria was shorter in UCMSCs infusion group than that in UCMSCs non-infusion group [12(9-17) vs 17(12.0-26.5) day] and the difference was statistically significant ( P=0.045). And the duration of urinary tract irritation symptoms was shorter in UCMSCs infusion group than that in UCMSCs non-infusion group [18(11-30) vs 27(18.0-35.5) days] and the difference was statistically significant ( P=0.048). Conclusions:Indicated for post-ALLO-HSCT HC, infusion of UCMSCs may significantly shorten the course of disease.Age ≤30 years, haploid transplantation and preconditioning with positive ATG, aGVHD and CMV-DNA may boost the risks of HC post-allo-HSCT.And aGVHD is an independent risk factor for HC after allo-HSCT.

Objective:To evaluate the safety and efficacy of bortezomib plus highdose melphalan (L-phenylalanine nitrogen mustard) (Bor-HDM) pretreatment regimen for multiple myeloma (MM) with autologous hematopoietic stem cell transplantation (ASCT).Methods:From August 2008 to December 2021, the relevant clinical data were retrospectively reviewed for 58 MM patients undergoing MM transplantation.The conditioning regimens were Bor-HDM (n=36) and HDM (n=22). Non-hematopoietic adverse reactions, hematopoietic reconstruction time, remission rate post-ASCT and minimal negative rate of residual disease (MRD) on flow cytometry within 3 months post-ASCT and survivals were analyzed.Results:In Bor-HDM and HDM groups, median time of neutrophil engraftment was 12(8-30) and 11(8-29) day and median time of platelet reconstitution 16(8-33) and 16(7-32) day respectively.There was no significant inter-group difference ( P=0.890, P=0.638). In Bor-HDM group, the most common non-hematological adverse reactions were nausea (n=21, 58.0%) and diarrhea (n=11, 30.6%). There was no transplant-related death.Complete remission (CR) rate was (25/36, 69.4%) versus (9/22, 40.9%). The inter-group difference was statistically significant ( P=0.032). Median follow-up period was 29.0(2.0-91.0) vs. 20.5(5.0-114.0) month, 3-year progression-free survival(PFS)62.1% vs. 39.7% and 3-year overall survival(OS) 83.8% vs. 62.5%.There were relapse (n=10 vs.10) and death (n=6 vs. 7). Median PFS in Bor-HDM and HDM groups was non-attained and 27 months( P=0.047) and median OS time non-attained and 40 months respectively ( P=0.282). Multivariate analysis revealed that CR was an independent risk factor for PFS ( HR=28.896, 95% CI: 6.130-136.198, P<0.001). Non-CR was an independent risk factor for OS ( HR=3.843, 95% CI: 1.334-11.071, P=0.013; HR=28.595, 95% CI: 6.273-130.355, P<0.001). Conclusions:Bor-HDM pretreatment regimen of ASCT is both safe and efficacious for MM patients.