OBJECTIVETo investigate the differentiation of bone marrow stem cells in transplanted livers and its impact on the long-term survival of rats with orthotopic liver transplantation.

METHODSTwenty-four female recipient rats with orthotopic liver transplantation were randomized into blank-control group, D-hanks solution group, bone marrow stem cells group with postoperative infusion of stem cells, and the pathological changes of the liver grafts and survival time of the rats were observed. The differentiation of the bone marrow stem cells were assessed 60 days after transplantation using in situ hybridization histochemistry for Sry gene and alpha-fetoprotein (AFP) immunohistochemistry.

RESULTSIn rats with postoperative infusion of bone marrow stem cells through the portal vein, the median long-term graft survival time exceeded 180 days, significantly longer than that in the other two groups (P<0.05), and no obvious evidence of acute rejection was observed with positive Sry expression and AFP expression.

CONCLUSIONInfusion of bone marrow stem cells through the portal vein following liver transplantation may alleviate acute graft rejection and promote long-term liver graft survival and AFP expression.

Animals ; Cell Differentiation ; Female ; Graft Rejection ; prevention & control ; Graft Survival ; Hematopoietic Stem Cells ; cytology ; Liver ; pathology ; Liver Transplantation ; Portal Vein ; Rats ; Rats, Wistar

OBJECTIVETo discuss the causes and countermeasures of death of severe obstructive sleep apnea hypopnea syndrome (OSAHS) patients undergoing tests or treatment.

METHODSA retrospective study of the data of six patients with severe OSAHS who died undergoing tests or treatment in 6 hospitals was conducted.

RESULTSAmong the 6 patients, overly fat were found in 2 cases, hypertension in 2 cases, cardiac diseases in 2 cases, abnormal pulmonary function or chest X-ray in 2 cases, diabetes in 2 cases, while 2 cases had no above positive finding. Three patients did not have polysomnography (PSG). One patient died during PSG test. Two patients' apnea hypopnea index (AHI) were 56 times/h and 82 times/h respectively. The causes of death were as follows, two patients died of airway obstruction and unsuccessful tracheal intubation before uvulopalatopharyngoplasty (UPPP) operation. One died of encephaledema in long-term coma due to airway obstruction, long time poor oxygenation and unsuccessful tracheal intubation with muscle relaxants before hemostasis procedure for treating postoperative hemorrhage. One patient whose preoperative pulmonary function showed increased airway resistance died of cardiac and respiratory arrest after tracheal extubation when UPPP surgery finished. One who was found to have potential cardiac and pulmonary disorder died of cardiac arrest when doctors were replacing his tracheal intubation with a cannula, before the tube replacing procedure the patient was conscious. One who had hypertension, coronary heart disease, diabetes mellitus and adiposis suffering from respiratory acidosis with his electrocardiogram showing a level-three heart function died of cardiac arrest with repeated apnea during PSG monitoring.

CONCLUSIONSOSAHS patients with severe complications should be treated extremely carefully. The indication of surgery should be strictly observed. OSAHS patients can not undergo surgery until their cardiopulmonary functions and general conditions are improved.

Adult ; Cause of Death ; Female ; Humans ; Male ; Middle Aged ; Retrospective Studies ; Sleep Apnea, Obstructive ; mortality

Objective To research the changes of clinical symptoms and serum free radical in children with severe Henoch-Sch(o)nlein purpura (HSP) between before and after accepted hemoperfusion.And evaluate the curative effect of hemoperfusion treatment for severe HSP and discuss the mechanism.Methods Twenty-three severe HSP patients in Children's Medical Center of the Second Xiangya Hospital of Central South University which were divided into 2 groups:13 cases were divided into traditional treatment group,10 cases were divided into hemoperfusion group; 11 healthy children were divided into healthy control group.The alleviate situation of clinical symptoms were observed and recorded such as purpura,abdominal pain,joint pain,hematuria,albuminuria,and the changes of urine RBC count,24 hour urine protein quantitative before and after hemoperfusion and traditional treatment.Collected the serum before and after the first time hemoperfusion treatment,after the second time hemoperfusion treatment,after the third time hemoperfusion treatment in hemoperfusion group;Collected the serum before and after conventional therapy in traditional treatment group; Collected only once serum in healthy control group.And then their superoxide anion(O2-·),hydroxy radical(· OH),hydrogen dioxide(H2O2),malonaldehyde(MDA) values were detected with spectrophotometry.The differences of each index among each group were compared respectively.Results After treatment,the clinical symptoms of 23 severe HSP children such as rash,abdominal pain,joint pain,hematuria,albuminuria which were reliefed compared to before treatment.The urine erythrocyte count,24 hour urine protein quantitative were reduced in 2 groups,but the symptoms of perfusion group children relieved faster,and the clinical index decreased more obviously.The serum O2-·,· OH,H2O2,MDA levels of 23 HSP children were significantly higher than those of the healthy control group (all P ＜ 0.01).In the traditional treatment group,compared between before and after treatment,the indexes were decreased (all P ＜ 0.05) ; After 3 times of hemoperfusion,all indexes were decreased in hemoperfusion group,but only after the third hemoperfusion the indexes were decreased statistically significant(all P ＜ 0.05).But compared with the hemoperfusion group,the index were decreased more apparently in after the third hemoperfusion,and it was statistically significant (P ＜ 0.05).Conclusions 1.The serum free radicals are increased in severe HSP children,they may play a role in vasculitis.2.For severe HSP,the recent therapeutic effect of hemoperfusion ally with traditional treatment is better than the alone traditional treatment.3.Hemoperfusion ally with traditional treatment can remove more effectly the serum free radicals,reduce lipid peroxide products,then mitigate the damage of the oxidate stress to the vascular endothelial.

OBJECTIVETo detect the VHL gene mutations in a Chinese family with nonsyndromic pheochromocytoma.

METHODSMutations of VHL gene were detected in a Chinese family with nonsyndromic pheochromocytoma. Five patients and fifteen relatives were involved in this study. Peripheral blood was collected and total genomic DNA was prepared for polymerase chain reaction (PCR). PCR products of all the three exons of VHL gene were purified and a direct gene sequence analysis was performed.

RESULTSAll the five patients presented a codon 125 from Histidine (H) to Proline (P) change at nucleotide 587 (A --> C) in exon 2. Seven members of fifteen relatives were carriers with the same VHL gene mutation. Two carriers were detected with bilateral adrenal tumors and right renal cyst respectively by ultrasonic inspection.

CONCLUSIONThe novel VHL gene mutation detected in this kindred may be the causative gene. Genetic test can detect the carriers in an early period. It is recommended as a routine method of genetic test in nonsyndromic pheochromocytoma patients.

Adolescent ; Adrenal Gland Neoplasms ; diagnosis ; ethnology ; genetics ; Adult ; Asian Continental Ancestry Group ; genetics ; Base Sequence ; Child ; China ; DNA Mutational Analysis ; Family Health ; Female ; Genetic Testing ; Humans ; Male ; Mutation ; Pedigree ; Pheochromocytoma ; diagnosis ; ethnology ; genetics ; Polymerase Chain Reaction ; Von Hippel-Lindau Tumor Suppressor Protein ; genetics ; Young Adult

BACKGROUND AND OBJECTIVECytokine-induced killer (CIK) cells have high anti-tumor activity for hepatocellular carcinoma (HCC). Whether CIK cell therapy can eradicate residual cancer cells and prevent or postpone tumor relapse after transcatheter arterial chemoembolization (TACE) should be testified. This study was to evaluate the efficacy of CIK cell therapy combined with TACE on HCC.

METHODSA total of 146 consecutive patients with unresectable HCC were divided into combination group (72 patients treated with CIK cell therapy combined with TACE) and TACE group (74 patients treated only with TACE). The progression-free survival (PFS) and overall survival (OS) were analyzed.

RESULTSThe 6-month, 1-year, and 2-year PFS rates were 72.2%, 40.4%, 25.3% in combination group, and 34.8%, 7.7%, 2.6% in TACE group. The median time to progression was 11 months [95% confidence interval (CI), 8-14 months] in combination group and 5 months (95% CI, 4-7 months) in TACE group. The estimated 6-month, 1-year, and 2-year OS rates were 90.3%, 71.9%, 62.4% in combination group, and 74.6%, 42.8%, 18.8% in TACE group. The median OS was 31 months (95% CI, 27-35 months) in combination group and 10 months (95% CI, 7-13 months) in TACE group. The times of TACE, ECOG performance status, and CIK cell therapy were independent prognostic factors for PFS and OS.

CONCLUSIONAdjuvant immunotherapy with CIK cells could greatly improve the efficacy of TACE on HCC, and plays an important role in prolonging the PFS and OS of HCC patients after TACE.

Carcinoma, Hepatocellular ; pathology ; therapy ; Chemoembolization, Therapeutic ; methods ; Combined Modality Therapy ; Cytokine-Induced Killer Cells ; transplantation ; Disease-Free Survival ; Female ; Humans ; Liver Neoplasms ; pathology ; therapy ; Male ; Middle Aged ; Proportional Hazards Models ; Remission Induction ; Survival Rate

OBJECTIVETo investigate the association between a new polymorphism (IVS3-20 T>C GenBank accession number: AY463003) in intro 3 of the parkin gene and the risk for Parkinson's disease (PD) in Chinese, particularly the relation between this polymorphism and the age of onset of PD patients.

METHODSPD was diagnosed according to the criteria of Core Assessment Program for Intracerebral Transplantations(CAPIT). All patients and controls were examined by two neurologists and were of the Han ethnic background. Polymerase chain reaction (PCR), denaturing high performance liquid chromatography (DHPLC) and sequencing were used to determine the genotype of each subject.

RESULTSA total of 312 PD patients (including 99 early-onset PD patients and 213 late-onset PD patients) and 236 controls were studied. The C/C homozygote was not found in this study. Chi-square analysis revealed that the frequencies of the C allele and T/C genotype were higher in total PD group but were not statistically different from those of the control group (P=0.6350 and 0.6331, respectively). After being stratified by age of onset, the frequency of T/C genotype was significantly higher (OR=3.52, 95%CI 0.97-13.13) in PD group with an onset age at or below 45 years old (7.07%), compared with that in the control group (2.12%). Similarly, C allele was much higher (OR=3.42, 95%CI 0.96-12.57, P=0.0276) in the early-onset PD group (3.90%) than that in the control group (1.06%). The linear trend analysis showed that both the T/C genotype and C allele increased significantly in the PD group with the increase of the onset age [chi-square(trend of Genotypes)=4.414, P=0.036; chi-square(trend of Alleles)=4.344, P=0.037]. On the other hand, there was no difference in the frequencies of allele and genotype between the late-onset PD patients and controls.

CONCLUSIONThe above results suggest that the parkin IVS3-20 T>C polymorphism might be a genetic risk factor for early-onset PD in Chinese.

Adult ; Age Distribution ; Aged ; Aged, 80 and over ; Base Sequence ; Female ; Humans ; Male ; Middle Aged ; Molecular Sequence Data ; Parkinson Disease ; genetics ; Polymorphism, Genetic ; Sex Distribution ; Ubiquitin-Protein Ligases ; genetics

This study was aimed to explore the clinical significance of monitoring level of minimal residual disease (MRD) at different time point in B-lineage childhood acute lymphoblastic leukemia (B-ALL). Two hundred and six children with B-ALL were enrolled in this study from Augest 2008 to September 2011 in our hospital. MRD levels were detected by flow cytometry at day 15, 33 and week 12 after initial chemotherapy. The event-free survival (EFS) for patients based on MRD levels measured at different stages of chemotherapy were compared by Kaplan Meier analyses. The results showed that out of 206 cases 196 cases achieved complete remission (CR) after induction therapy (CR rate 95.1%), the 1- and 3-year EFS rate were (92.7 ± 1.8)% and (78.7 ± 3.7)%, respectively, and the 3-year EFS rate was (85.6 ± 4.9)% in standard risk group, (82.1 ± 5.8)% in intermediate risk group and (58.1 ± 9.2)% in high risk group, there was significant statistical difference between above mentioned 3 groups (P < 0.001). The MRD analysis at different time points showed that the higher the MRD level, the lower the 3-year EFS rate of children with ALL, in which the 3-year EFS rate of MRD ≥ 10(-2) at day 15, MRD ≥ 10(-3) at day 33 and MRD ≥ 10(-3) at week 12 were significantly lower. The MRD ≥ 10(-3) at week 12 was proven to be an independent predictor by multivariate Cox proportional-hazards regression model. The 3-year EFS rate for patients with MRD < 10(-3) and MRD ≥ 10(-3) at week 12 were (86.3 ± 4.1)% vs (55.8 ± 9.1)% (P < 0.05); 8 relapsed among 98 cases with negative MRD (MRD < 10(-4)) at day 33, 19 relapsed among 108 cases with positive MRD at day 33 between the two groups for recurrence rate has significant difference (P < 0.05). It is concluded that dynamically monitoring MRD by multi-parameter flow cytometry can precisely evaluate treatment response, judge treatment outcome and predict relapse in childhood B-ALL. The MRD 10(-2) at day 15, MRD 10(-3) at day 33 and MRD 10(-3) at week 12 should be considered as the best cut-off. MRD ≥ 10(-3) at week 12 was proven to be an independent factor of poor prognosis.
Child ; Child, Preschool ; Female ; Flow Cytometry ; methods ; Humans ; Infant ; Male ; Neoplasm, Residual ; diagnosis ; therapy ; Precursor Cell Lymphoblastic Leukemia-Lymphoma ; diagnosis ; therapy ; Prognosis ; Retrospective Studies

OBJECTIVETo express the Gag protein of HIV-1 strain CN54 in Pichia pastoris (P.pastoris), optimize fermentation parameters and purify Gag antigen.

METHODSThe Gag gene was subcloned into downstream of aox1 promoter of Pichia expression vector pPS1.0, an integrative vector which possesses an identical 5' untranslated region as the natural aox1 gene and employs both in vitro construction and in vivo selection for multi-copy integrants. The recombinant vector was introduced into P.pastoris strain GS115 by electroporation and selected with G418 for Gag gene integration. Super G418 resistant clones were selected and screened for Gag expression. The engineered P.pastoris was cultured to high cell density (>300 A600 Units/ml) in a 5L fermentor. Through methanol induction, the expression level of Gag reached 120 mg/L. Intracellularly expressed Gag was released by high-pressure homogenization and purified through Sepharose FF and DEAE Sepharose FF column chromatography, the purity of Gag reached up to 90%.

RESULTSWestern-blotting suggested that purified Gag expressed in P.pastoris could react specifically with serum from HIV infected individual.

CONCLUSIONGag antigen expressed in P.pastoris has provided a good basis for the development of a new generation of HIV vaccine candidates against some Chinese prevalent strains.

Blotting, Western ; Electrophoresis, Polyacrylamide Gel ; Fermentation ; Gene Expression ; HIV Core Protein p24 ; biosynthesis ; genetics ; isolation & purification ; HIV-1 ; genetics ; metabolism ; Humans ; Pichia ; genetics ; metabolism

PURPOSE: The purpose of this study was to subdivide M1 stage nasopharyngeal carcinoma (NPC) patients with bone-only metastases for prognosis prediction while identifying the treatment effect of locoregional radiotherapy (LRRT) and metastasis radiotherapy (MRT) among patients with different risk. MATERIALS AND METHODS: From November 2006 to October 2016, a total of 226 patients with bone-only metastasic NPC were retrospectively enrolled. All patients developed distant lesions before receiving treatment. All potential prognostic factors were considered and the correlation of the M1 subdivisions with overall survival (OS) was determined by Cox regression hazards model. Kaplan–Meier curves were used to appraise survival condition and log-rank testing was used to compare the differences. RESULTS: The median follow-up time was 33.9 months (range, 3 to 126 months). According to multivariate Cox proportional hazard analysis, the number of metastatic lesions and Epstein-Barr virus (EBV) DNA status after palliative chemotherapy (PCT) were independent prognostic factors for OS. Thus, we subdivided patients into three risk groups according to these two factors. Systemic chemotherapy combined with LRRT may benefit patients in low- and intermediate-risk groups but not in the high-risk group. Further aggressive MRT based on systemic chemotherapy showed no survival benefit in any risk group. CONCLUSION: The stratification of NPC patients with bone-only metastasis based on EBV DNA after PCT and the number of metastatic lesions provided promising prognostic value and could aid clinicians in person-specific treatment.
Diagnosis ; DNA ; Drug Therapy ; Follow-Up Studies ; Herpesvirus 4, Human ; Humans ; Neoplasm Metastasis ; Prognosis ; Proportional Hazards Models ; Radiotherapy ; Retrospective Studies

To unravel the relation of HLA-DRB1*15 with childhood acute lymphoblastic leukemia (ALL), 162 childhood patients with ALL were selected for this investigation. 1 000 normal umbilical cord blood samples were used as control.HLA-DRB1*15 and HLA-DRB5* were typed by polymerase chain reaction (PCR) analysis. The relation of HLA-DRB1*15 with childhood ALL was studied by calculating the chi-square test and relative risk. The results showed that the antigen frequencies and allele frequencies of HLA-DRB1*15 in childhood patients with ALL were 40.12% and 22.62% respectively, while the antigen frequencies and allele frequencies of HLA-DRB1*15 in control were 30.80% and 16.81% respectively, there were significant difference between them (chi(2) = 5.560, p = 0.018, RR = 1.506). In conclusion, the antigen frequencies and allele frequencies of HLA-DRB1*15 in childhood patients with ALL were higher than those in control, so the HLA-DRB1*15 gene is one of the genetic risk factors for childhood ALL. These preliminary data may be useful for further study on the pathogenesis of childhood ALL.
Adolescent ; Case-Control Studies ; Child ; Child, Preschool ; Female ; Gene Frequency ; HLA-DR Antigens ; genetics ; HLA-DRB1 Chains ; Humans ; Infant ; Male ; Precursor Cell Lymphoblastic Leukemia-Lymphoma ; genetics