Objective To summarize the timing and diagnostic value of endoscopic biopsy of intestinal graft after small bowel transplantation (SBTx).Methods Fifteen cases of SBTx were divided into 3 eras:era Ⅰ (1994-1995)-3 cases of SBTx treated with cyclosporine-based immunosuppression; era Ⅱ (2003-2006)-7 cases of SBTx treated with tacrolimus-based immunosuppression,and era Ⅲ (2007-present) than CVC group 5 cases of SBTx treated with Atemtuzumab induction therapy and maintenance tacrolimus monotherapy.The scheme of endoscopic surveillance was initially used in era Ⅲ, the first endoscopic biopsy was performed on postoperative day 3,2 times weekly during the first month, followed once weekly during months 2-3, once every other week during months 4-6 and once monthly thereafter.When clinical signs and symptoms of rejection were present, and during rejection episodes, the additional endoscopic biopsies were also performed.Results A total of 276 biopsies of these 15 SBTx recipients were obtained.Fifty-one biopsies (18.5%) were diagnosed as acute cellular rejection (ACR), which included IND to mild (n = 32,11.6 %), moderate (n = 9,3.3 %), and severe (n = 1 0, 3.6 %), two biopsies (0.7 %) were diagnosed as cytomegalovirus (CMV) enteritis and other 2 biopsies (0.7 %) bacteria enteritis.The ACR episodes verified by biopsy pathology and undergoing anti-rejection treatment were 20 (11 IND to mild,5 moderate,and 4 severe) ,and 1 episode of CMV enteritis and 1 episode of bacteria enteritis were observed.Conclusion Endoscopic surveillance and biopsy pathology is crucial diagnostic tool for ACR and sepsis.ACR surveillance after SBTx and early diagnosis of ACR could be made with scheming endoscopic biopsies.Endoscopic biopsy can be used to make differential diagnosis when clinical signs and symptoms were present, and to guide the treatment during anti-rejection episode.

Objective Cytomegalovirus (CMV) has remained the most significant pathogen that threatens the outcome of small bowel transplantation (SBTx). This paper To outline preliminary experience of prophylaxis and treatment of cytomegalovirus (CMV) in 15 cases subject to small bowel transplantation (SBTx) and also review current progress of diagnosis and treatment of CMV.Methods Fifteen cases of SBTx were divided into 3 eras: era Ⅰ (1994-1995)-3 SBTx treated with cyclosporine-based immunosuppression; era Ⅱ (2003-2006)-7 SBTx treated with tacrolimus-based immunosuppression; and era Ⅲ (2007-present)-5 SBTx treated with Alemtuzumab induction therapy and maintenance tacrolimus monotherapy. No antiviral prophylaxis after SBTx was applied during era Ⅰ; in era Ⅱ, ileoscopic and pathological diagnosis of CMV graft enteritis was defined, and plasma diagnosis tools including CMV-IgM, CMV pp65 and CMV DNA with PCR were introduced. 2-3 weeks intravenous ganciclovir prophylaxis of CMV was underway, followed by 3 months oral acyclovir; In era Ⅲ, more precise real-time PCR technique was used to detect CMV DNA copies, and the schedule of the CMV surveillance was set up, antiviral prophylaxis therapy was modified to 2-3 weeks intravenous ganciclovir and 3 months oral ganciclovir, and preemptive therapy to halt the progression of asymptomatic infection to clinical disease was also introduced.Results Two of 15 SBTx recipients suffered from CMV with the occurrence rate of 13.3%. One recipient in era Ⅱ suffered from CMV graft enteritis on postoperative day 45, and CMV pneumonia on postoperative day 64, he received intravenous ganciclovir and thymus peptide, paused tacrolimus maintenance, and finally he died from severe acute cellular rejection. 94 100 copies/ml of CMV DNA in periphery blood of a recipient in era Ⅲ was detected with real-time PCR at 3rd month after SBTx, and a preemptive therapy successfully halted the CMV infection.Conclusion Antiviral prophylaxis therapy and close surveillance of CMV infection after SBTx should be performed, and preemptive therapy can also halt the CMV infection. When CMV disease occurs, the recipient should receive effective antiviral therapy, and acute cellular rejection also should be closely monitored at same time.

Objective The protective effect of hydrogen sulfide (H2 S) against myocardial ischemia/reperfusion ( IR) injury via anti-apoptotic signaling is well established , but the underlying mechanism remains unclear .This study was to investigate whether H 2 S could protect cardiomyocytes from endoplasmic reticulum stress ( ERS)-mediated apoptosis in hypoxia/reoxygenation ( HR) injury by regulating the expression of microRNA-455 ( miR-455 ) . Methods Cardiomyocytes from neonatal SD rats were primarily cultured and the model of HR injury was established .The cardiomyocytes were divided into a control group (normally cultured for 27 hours), an HR group (subjected to HR injury), and an H2S protection group (pretreated with the precursor of H2S NaHS at 40 μmol/L at 30 min before HR treatment followed by the same procedure as in the HR group ) .The cell viability was monitored by MTT , the release of lactate de-hydrogenase ( LDH) in the culture supernatant measured by full-automatic chemical analysis , and the apoptosis rate of the cardiomyo-cytes detected by flow cytometry .The mRNA and protein expressions of Grp 78 and caspase-12 were determined by real-time RT-PCR and Western bot .To verify whether miR-455 was involved in the ERS-mediated apoptosis of the cardiomyocytes , the cells were subjec-ted to HR after transfected with miR-455 mimic or anti-miR-455 oligonucleotide (AMO) for 24 hours, followed by detection of the ex-pressions of Grp78 and caspase-12. Results After HR injury, the H2 S protection group showed an enhanced viability of the cardio-myocytes in comparison with the control group ([67.02 ±6.90] vs [29.27 ±5.66] %), an decreased LDH release ([91.33 ± 10.63] vs [168.17 ±15.38] U/L), and a reduced rate of cell apoptosis ([13.98 ±1.90] vs [24.31 ±2.79] %).H2 S pretreat-ment significantly downregulated the mRNA and protein expressions of Grp 78 and caspase-12 (1.66 ±0.39 vs 2.56 ±0.34;1.75 ± 0.32 vs 2.54 ±0.48;2.01 ±0.45 vs 3.26 ±0.34;1.85 ±0.52 vs 3.21 ±0.84, P<0.05).The mRNA and protein expressions of Grp78 and caspase-12 were evidently increased after transfection with miR-455 mimic (3.56 ±0.37 vs 1.00 ±0.00;3.61 ±0.41 vs 1.00 ±0.00;2.87 ±0.38 vs 1.00 ±0.00;2.98 ±0.49 vs 1.00 ±0.00), but remarkably decreased after transfection with miR-455 AMO (0.62 ±0.16 vs 1.00 ±0.00;0.65 ±0.13 vs 1.00 ±0.00;0.54 ±0.13 vs 1.00 ±0.00;0.62 ±0.16 vs 1.00 ±0.00, P<0.05). Conclusion H2S could protect cardiomyocytes from HR injury by regulating the expression of miR-455 and reducing ERS-mediated cell apoptosis .

To explore a new way to treat CML, inhibitory effect of small interfering RNA (SiRNA) on bcr-abl fusion gene expression of K562 cell line was studied. SiRNA for bcr-abl gene was designed and transfected into K562 cells, bcr-abl gene expression was tested by RT-PCR. The results showed that bcr-abl gene expression was inhibited by using siRNA in dose-dependent manner and reduced to 19.9% and 26.6% of the control at 24 and 48 hours after transfection with 0.2 micro g siRNA respectively. K562 cells proliferation was suppressed finally, but bcr-abl gene expression restored at 72 hours. In conclusion, anti-bcr-abl siRNA can effectively inhibit bcr-abl gene expression of K562 cell line.
Dose-Response Relationship, Drug ; Genes, abl ; Humans ; K562 Cells ; metabolism ; RNA, Messenger ; analysis ; RNA, Small Interfering ; pharmacology ; Transfection

Because of irregular shapes of Chinese herbal pieces, we simplified the previously deduced general extraction kinetic model for TCMs, and integrated particle diameters of Chinese herbs that had been hard to be determined in the final parameter "a". The reduction of the direct determination of particle diameters of Chinese herbs was conducive to increase the accuracy of the model, expand the application scope of the model, and get closer to the actual production conditions. Finally, a simplified model was established, with its corresponding experimental methods and data processing methods determined. With total flavonoids in Scutellariae Radix as the determination index, we conducted a study on the adaptability of total flavonoids extracted from Scutellariae Radix with the water decoction method in the model. The results showed a good linear correlation among the natural logarithm value of the mass concentration of total flavonoids in Scutellariae Radix, the time and the changes in the natural logarithm of solvent multiple. Through calculating and fitting, efforts were made to establish the kinetic model of extracting total flavonoids from Scutellariae Radix with the water decoction method, and verify the model, with a good degree of fitting and deviation within the range of the industrial production requirements. This indicated that the model established by the method has a good adaptability.
Chemical Fractionation ; methods ; Drugs, Chinese Herbal ; isolation & purification ; Flavonoids ; isolation & purification ; Kinetics ; Models, Theoretical ; Scutellaria baicalensis ; chemistry ; Water ; chemistry

Adolescent ; Adult ; Bleomycin ; therapeutic use ; Child ; Child, Preschool ; Female ; Hemangioma ; drug therapy ; Humans ; Laryngeal Neoplasms ; drug therapy ; Male ; Middle Aged ; Pharyngeal Neoplasms ; drug therapy ; Young Adult

BACKGROUNDGreat advances have been made in the diagnosis, molecular pathogenesis and treatment of acute lymphoblastic leukemia (ALL) in the past decade. Due to the lack of large population-based studies, the recent trends in the incidence and geographic variations of ALL in Shanghai, China have not been well documented. To better understand the incidence and epidemiological features of ALL in Shanghai, we conducted a retrospective survey based on the database from the Shanghai Center for Disease Control and Prevention (CDC) and the medical records in all large-scale hospitals in Shanghai, especially those 30 major hospitals with hematology department.

METHODSAccording to the data from Shanghai CDC, 544 patients, with a median age of 32 years (ranging 1.2 - 89 years), were diagnosed as de novo ALL from January 1, 2002 to December 31, 2006, and they were followed up until December 31, 2007.

RESULTSThe average annual incidence of ALL in Shanghai was 0.81/100 000. The incidence in men (0.86/100 000) was slightly higher than that in women (0.75/100 000). The age-stratified incidence showed that the incidence was 2.31/100 000 in patients ≥ 17 years old, 0.54/100 000 in those 18 - 34 years old, 0.46/100 000 in those 35 - 59 years old, and 0.94/100 000 in those ≥ 60 years old. Moreover, there were substantial geographic variations in the incidence of ALL, with the incidence in Chongming county, an island in the east of Shanghai city being 0.60/100 000, much lower than those of other districts. Both French-American-British (FAB) and World Health Organization (WHO) classification systems were applied in the present study. Eighty-eight patients were diagnosed as L1 (26.2%), 193 L2 (57.4%), and 55 L3 (16.4%). For 302 patients with immunophenotypic results, 242 were identified as B cell origin (80.1%), 59 as T cell origin (19.5%), and 1 as biphenotype (0.4%). The leukemia cells in 61 patients co-expressed one or two myeloid antigen (20.2%). For 269 patients with cytogenetic results, the incidences of t(9;22) in patients aged < 10, 11 - 17, 18 - 44, 45 - 59 and ≥ 60 years old were 4.2%, 11.4%, 19.2%, 23.1% and 5.3%, respectively.

CONCLUSIONCompared with the previous data, the incidence of ALL is increased in Shanghai, and has a geographic distribution characteristic.

Adolescent ; Adult ; Aged ; Aged, 80 and over ; China ; epidemiology ; Data Collection ; Female ; Humans ; Incidence ; Male ; Middle Aged ; Precursor Cell Lymphoblastic Leukemia-Lymphoma ; epidemiology ; Young Adult

OBJECTIVETo avoid disadvantages of two-stage cancellus bone autograft, we investigated the feasibility of one-stage allograft for reconstructing the bone defect resulting from debridement of chronic osteomyelitis in limbs.

METHODSBetween Feb. 1999 and Apr. 2004, 35 cases of chronic osteomyelitis (8 cases of nonunion) underwent one-stage allograft after debridement in our hospital.

RESULTSThirty-five cases were followed up for an average period of 28 months (range, 13 to 55 months), in which 32 cases (91.43%) were found no infection, and 3 cases (8.57%) were confirmed recurrence of infection. Four out of 8 cases of bone nonunion healed in 9.5 months on average (range, 3 to 12 months), and another case also acquired union after redebridement and autograft of ilium due to infection recurrence 35 days after surgery. Renonunion occurred in 3 cases, 2 out of whom healed after secondary operation with autograft. One case of renonunion and 2 cases of infection recurrence refused further treatment.

CONCLUSIONSA high rate of infection arrest can be attained when one-stage allograft is used to reconstruct the bone defect of chronic osteomyelitis after debridement in limbs. Therefore, chronic osteomyelitis should not be regarded as a contraindication to one-stage allogeneic bone grafting. Renonuion, however, achieves a relatively high rate, especially in cases of segmental bone defect.

Adolescent ; Adult ; Aged ; Bone Transplantation ; methods ; Child ; Child, Preschool ; Chronic Disease ; Debridement ; Female ; Humans ; Male ; Middle Aged ; Osteomyelitis ; surgery ; Transplantation, Homologous

OBJECTIVETo explore the effects and mechanism of CD4+ CD25+ regulatory T cells (Tregs) in mouse experimental colitis treated by CLYSTER No. 1.

METHODThe mouse model of experimental colitis was established by dinitrochlorobenzene (DNCB)-acetic acid (AA) in mice DNCB and AA. Adult KM mouse were randomly divided into four groups: normal control group, experimental colitis model group, SASP and Chinese medicine therapeutic groups. Proportion of CD4 CD25+ Tregs in peripheral blood (PB) and mesenteric lymph node (MLN) was estimated by flow cytometry at the end of one or two week after treating with SASP and CLYSTER No. 1.

RESULTThe model of experimental colitis in mouse was successfully established. Compared with normal control group, the proportion of CD4 CD25 Tregs was markedly decreased in PB and MLN of model control group of experimental colitis. But it was significantly increased in therapeutic groups of SASP and CLYSTER No. 1, and their CD4+ CD25+ Tregs in PB and MLN were much more than the model control group at the end of one or two weeks after treating with SASP and CLYSTER No. 1.

CONCLUSIONCD4+ CD25+ Tregs with strong immune suppression could play a central role in the initiation and development of mouse experiment colitis, and the CLYSTER No. 1 might exert its therapeutic effects on UC by the regulation of number and function of CD4+ CD25+ Tregs.

Animals ; CD4 Antigens ; immunology ; Colitis ; immunology ; Disease Models, Animal ; Drugs, Chinese Herbal ; pharmacology ; Female ; Flow Cytometry ; Interleukin-2 Receptor alpha Subunit ; immunology ; Male ; Mice ; Random Allocation ; T-Lymphocytes, Regulatory ; drug effects ; immunology

OBJECTIVETo examine the association between CD133 expression and invasion of gastric cancer, and to elucidate whether CD133 can promote the invasion and metastasis of gastric cancer via epithelial-mesenchymal transition (EMT).

METHODSThe CD133(+) and CD133(-) KATO-III( cells were sorted by magnetic activated cell sorting (MACS). The invasion ability was detected by Transwell method. RT-PCR and Western blot were used to detect the expression of EMT-related factors in KATO-III( cells before and after CD133 was knocked out by siRNA method. The expressions of CD133 and EMT-related proteins of cancer and adjacent normal tissues in 50 patients with gastric cancer were detected by Western blot, and correlations among protein expressions were also analyzed.

RESULTSAs compared to CD133(-) cells, the number of broken-membrane cells was significantly higher (67.7±10.5 vs. 13.3±6.8, P=0.001) and the invasion ability was stronger (P<0.05) in CD133(+) cells, while the mRNA expression levels of Snail and N-cadherin were significantly higher in CD133(+) cells (0.311±0.015 vs. 0.223±0.016, P=0.040; 0.581±0.020 vs. 0.270±0.018,P=0.004), and the protein expression levels of Snail and N-cadherin were significantly higher in CD133(+) cells as well (0.513±0.015 vs. 0.179±0.023, P=0.030; 0.538±0.028 vs. 0.202±0.032, P=0.020), but E-cadherin mRNA and protein levels were significantly lower in CD133(+) cells (0.231±0.009 vs. 0.460±0.015, P=0.040; 0.426±0.030 vs. 0.748±0.027, P=0.040). After CD133 knock-out, the expressions of Snail and N-cadherin were down-regulated (P<0.05) and the expression of E-cadherin was up-regulated (P<0.05). As compared to normal mucosal tissues, the protein expression levels of Snail, N-cadherin and CD133 in gastric cancer tissues were significantly higher(0.635±0.119 vs. 0.485±0.116, P=0.029; 0.599±0.114 vs. 0.259±0.108, P=0.020; 0.754±0.154 vs. 0.329±0.134, P=0.001), while the protein expression of E-cadherin in gastric cancer tissues was lower (0.378±0.123 vs. 0.752±0.156, P=0.003). The protein expressions of Snail and N-cadherin were positively correlated with CD133 expression (r=0.278, P=0.048; r=0.406, P=0.003) and the protein expression of E-cadherin was negatively correlated with CD133 expression (r=-0.504, P=0.000).

CONCLUSIONCD133(+) cells in primary lesion of gastric cancer have relatively higher invasion ability, which may promote the metastasis of gastric cancer via up-regulation of EMT-related factors.

AC133 Antigen ; Adult ; Aged ; Aged, 80 and over ; Antigens, CD ; metabolism ; Cell Line, Tumor ; Epithelial-Mesenchymal Transition ; Female ; Glycoproteins ; metabolism ; Humans ; Male ; Middle Aged ; Neoplasm Invasiveness ; Neoplasm Metastasis ; Peptides ; metabolism ; Stomach Neoplasms ; metabolism ; pathology