Volume 87  ·   Issue 30  ·  2007

At present, there is no uniform diagnostic standard for primary small hepatocellular carcinoma in the world. Most literatures define small hepatocellular carcinoma as the diameter of single or two adjacent tumor nodules and ≤5 cm, but some literatures also use the tumor diameter ≤3 cm as the boundary.

"flesh-eating bacteria" infection is a rapidly progressing skin disease, which can quickly destroy skin, fascia, muscle and other soft tissues, and cause tissue necrosis, toxic shock and multiple organ failure in a short period of time. The mortality and disability rate are extremely high.

Methods 251 cases of hepatocellular carcinoma liver transplantation in our hospital from April 2001 to February 2006 were retrospectively analyzed. The Kaplan-Meier survival rate was statistically analyzed. The influence of various tumor-related factors on the survival rate and tumor-free survival rate after hepatocellular carcinoma liver transplantation was analyzed. The difference between groups was tested by Log Rank, and the risk factors that had significant influence on the prognosis were analyzed by COX multivariate regression. However, there was no significant difference between preoperative treatment history and hepatitis background on postoperative survival rate and tumor-free survival rate (P>0.05). The indexes of Cox multivariate analysis that had significant influence on survival rate or tumor-free survival rate were: macrovascular tumor thrombus, tumor dissemination foci and microscopic tumor thrombus (P<0.05 or P<0.01). Conclusion Edmondson grade Ⅲ ~ Ⅳ, microscopic tumor thrombus, tumor located in left lobe or both lobes, tumor dissemination foci and preoperative AFP ≥300 μ g/L are all risk factors for liver transplantation of liver cancer, suggesting poor prognosis. The prognosis of macrovascular tumor thrombus and lymph node metastasis is extremely poor, so it should be listed as contraindication for liver transplantation of liver cancer.

Objective to investigate the changes of the sensitivity of hepatocellular carcinoma cells SMMC-7721 to fluorouracil chemotherapeutic drugs under the action of interferon-α (IFN-α) and its relationship with the expression of thymidine phosphorylase (TP). Methods the expression of TP mRNA in SMMC-7721 cells was detected by reverse transcription polymerase chain reaction (RT-PCR), and the sensitivity of hepatocellular carcinoma cells to fluorouracil chemotherapeutic drugs was detected by MTY assay. Results the upregulation of TP mRNA by IFN-α was time-dose dependent. The expression of TP mRNA in SMMC-7721 cells started to increase after 8 h of treatment with 10 000 U/ml IFN-α, reached a peak at 12 h, and then maintained at a high level to 72 h. The expression level of TP mRNA was significantly increased when IFN-α was treated at 5000 U/ml and 10 000 U/ml for 24 h, and the difference was statistically significant compared with that in the group without IFN-α treatment (P<0.05). IFN-α significantly increased the sensitivity of SMMC-7721 cells to 5 '-deoxy-5-fluorouridine (5, -dFUrd), and the IC50 of 5'-dFUrd decreased from (102.1±18.4) μ mol/L to (34.2±4.1) μ mol/L under the action of 10 000 U/ml IFN-α (P<0.05). However, the sensitivity of IFN-α to 5-fluorouracil (5-FU) was not significant, and the IC50 was (5.8±2.0) μ mol/L when 10 000 U/ml IFN-α was treated with 10 000 U/ml IFN-α. The increase of the sensitivity of 5'-dFUrd was positively correlated with the upregulation of TP mRNA by IFN-α, and the correlation coefficient was 0.6 (Pearson test P =0.008). Conclusion IFN-α significantly enhanced the effect of SMMC-7721 on 5-FU IFN-α upregulated TP mRNA expression in SMMC-7721 cells in a time-dose dependent manner.

Objective to study the protein markers associated with portal vein tumor thrombosis of hepatocellular carcinoma by comparative proteomics. Methods Two-dimensional gel electrophoresis (2-DE) was used to separate the total proteins from the primary tumor tissues of hepatocellular carcinoma and their portal vein tumor thrombosis tissues of the same patient. The differential proteins between the two groups were identified by mass spectrometry and protein database searching, and the differential protein spots were further detected and verified by Western blot. Results A total of 20 significant proteins were identified. Twelve proteins were expressed or significantly increased in the primary tumor tissues of hepatocellular carcinoma, including Galectin-1, high mobility group protein-1 (HMGBI), peroxiredoxin-1, Cyclophilin B, proliferating cell nuclear antigen (PCNA), etc. The results of 2-DE were verified by Western blot on the protein level for the high expression of Annexin V in portal vein tumor thrombus. Conclusion the formation of portal vein tumor thrombus in hepatocellular carcinoma is related to various proteins, among which the high expression of Annexin V may play an important role.

Clinical data A 48-year-old male was admitted to the hospital on December 7, 2005 due to fatigue, general distension and pain with low fever for more than 1 month. One month before admission, there was no obvious trigger for fatigue, general distension and pain, with low fever (no body temperature measured), nausea, anorexia, no vomiting, no fear of cold, chills, cough, expectoration, hemoptysis and other discomfort. Nasal bleeding occurred 3 days before admission, which stopped spontaneously 2 hours later.

Objective to observe the therapeutic effect of oxymatrine on hemorrhagic fever with renal syndrome. Methods 418 patients with hemorrhagic fever with renal syndrome were randomly divided into a treatment group and a control group. The treatment group was treated with oxymatrine intravenously, 600 mg once a day, for 7 days, and the control group was treated with traditional balanced salt therapy. The disease outcome and complications of the two groups were observed. The levels of serum urinary modulin (THP) and β 2 microglobulin (β 2M) in the patients' serum were dynamically detected, and the changes of serum interleukin-15 (IL-15) and soluble intercellular adhesion molecule (sICAM-1) levels were also observed. Results The course of disease and prognosis of the patients in the oxymatrine treatment group were shortened and improved. The levels of serum THP and β 2M were significantly lower than those in the control group, and the levels of serum IL-15 and sICAM-1 were also statistically significant compared with those in the control group after treatment.

Objective to investigate the changes of plasma osteoprotegerin (OPG) levels before and after levothyroid hormone (L-T4) treatment in patients with hypothyroidism (hypothyroidism). Methods 20 patients with clinical hypothyroidism, 20 patients with subclinical hypothyroidism and 20 healthy subjects (control group) were selected. All patients with hypothyroidism were treated with L-T4 supplementation until their thyroid function was within the normal range. Plasma OPG was measured by enzyme-linked immunoassay (ELISA). Results Before L-T4 treatment, the plasma OPG levels in clinical hypothyroidism group and subclinical hypothyroidism group were 3.13 ng/L ± 0.27 ng/L and 2.95 ng/L ± 0.24 ng/L, respectively, which were significantly higher than those in the control group (2.42 ng/L ± 0.26 ng/L, P =0.000). The results of multivariate correlation analysis showed that OPG was positively correlated with TSH (r =0.306, P<0.05) and negatively correlated with endothelium-dependent vasodilation (r = -0.675, P<0.01). After treatment to normal thyroid function, the plasma OPG levels in the two groups were significantly decreased (2.53 ng/L ± 0.28 ng/L, 2.54 ng/L ± 0.21 ng/L, P =0.000)

Objective to investigate the relationship between brachial artery endothelial function, carotid artery stiffness index and carotid intima-media thickness in the recovery period of Kawasaki disease. Methods A total of 204 subjects were enrolled in the trial, including 51 patients with Kawasaki disease complicated with coronary artery disease (coronary artery disease group), 50 patients with Kawasaki disease without coronary artery disease (normal coronary artery group) and 103 normal children (control group). Blood pressure, blood lipid, brachial artery endothelial function, carotid artery stiffness index and carotid intima-media thickness were observed. Results The brachial artery endothelial function was significantly lower in the coronary artery disease group (5.2% ± 1.9%) and the normal coronary artery group (6.8% ± 2.0%) than that in the control group (13.2% ± 4.1%, all P<0.01); The carotid artery stiffness index was significantly higher in the coronary artery disease group (4.0 ± 0.6) than that in the normal coronary artery group (3.6 ± 0.6) and the control group (3.4 ± 0.5, P<0.05 and P<0.01). Similarly, the carotid artery intima-media thickness in the coronary artery disease group (0.447 mm ± 0.024 mm) was significantly higher than that in the normal coronary artery group (0.426 mm ± 0.016 mm) and the control group (0.424 mm ± 0.016 mm, all P<0.01). Multiple regression analysis showed that age and group were the significant determinants of brachial artery endothelial function, carotid artery stiffness index and carotid artery intima-media thickness. Correlation test analysis showed that brachial artery endothelial function, carotid artery stiffness index and carotid artery intima-media thickness were significantly correlated in each group after adjusting for age. Conclusion There were vascular endothelial dysfunction, increased carotid artery stiffness and carotid artery intima-media thickening in the recovery period of Kawasaki disease

Objective to investigate the infection status of human herpesvirus 7 (HHV-7) in allogeneic hematopoietic stem cell transplantation (allo-HSCT) patients. Methods 680 peripheral blood samples from 72 allo-HSCT patients and 53 donor samples were collected. HHV-7 DNAemia was detected by nested polymerase chain reaction in donors and patients before and after transplantation. Results HHV-7 DNAemia was detected in 62 (86.1%) of 72 patients at least once after transplantation, with a median time of 15.6 (7-56) d. Twenty-nine patients (40.3%) had persistent HHV-7 DNAemia after transplantation. The incidence of persistent HHV-7 DNAemia was significantly higher in 48.9% (23/47) of non-congenital transplantation patients who received anti-thymocyte globulin (ATG) pretreatment regimen than in non-congenital transplantation patients [24% (6/25), P =0.040]. The incidence of persistent HHV-7 DNAemia after transplantation was significantly higher in 44.6% (39/65) of patients treated with glucocorticoid than in patients without glucocorticoid treatment [0% (0/7), P =0.037]. There was no significant correlation between HHV-7 and acute graft versus host disease, hemorrhagic cystitis, cytomegalovirus and human herpesvirus type 6 infection. Conclusion HHV-7 infection is very common in patients after allo-HSCT, and its persistent infection is closely related to the use of ATG and glucocorticoid after transplantation in pretreatment regimen

Objective to investigate the clinical significance of Janus kinase 2 (JAK2) V617F point mutation in patients with myeloproliferative disease (MPD). Methods Allele-specific polymerase chain reaction (AS-PCR) combined with sequencing was used to detect JAK2 V617F point mutation in peripheral blood mononuclear cells of 20 patients with chronic myeloid leukemia (CML), 23 patients with polycythemia vera (PV), 40 patients with essential thrombocythemia (ET), 8 patients with chronic idiopathic myelofibrosis (IMF) and 3 patients with hypereosinophilia syndrome (HES). Results JAK2 V617F mutation was found in 38 (51.4%) of 74 patients with BCR/ABL negative MPD, including 16 PV, 18 ET, 3 IMF and 1 HES. All positive samples and 10 negative samples were identified by sequencing, and the results were consistent. The percentage of hemoglobin, hematocrit and neutrophil in peripheral blood of JAK2 V617F mutation positive ET patients were significantly higher than those in negative group. Conclusion JAK2 V617F mutation positive ET patients

Objective to investigate the relationship between the allelic polymorphisms of-238G/A, -308G/A, -857C/T, -863C/A and serum TNF-α levels and chronic severe hepatitis B. Methods From December 2002 to December 2005, 98 patients with chronic severe hepatitis B and 211 patients with chronic hepatitis B were hospitalized in Beijing You'an Hospital. The allelic polymorphisms of-238G/A, -308G/A, -857C/T, -863C/A of TNF-α promoter were detected by polymerase chain reaction restriction fragment length polymorphism analysis. Results the alleles of-863C/A and-238G/A in the two groups (χ 2=0.61, P =0.436; χ 2=0.001, P =0.976) and the genotype frequencies were not significantly different (χ 2=1.16, P =0.552; χ 2=0.63, P =0.486). The frequency of-308 GA and-857 TT genotypes was significantly higher than that of chronic hepatitis B (χ 2=28.06, P =0.000; χ 2=19.69, P =0.000), and the difference was statistically significant. The genotypes of-308 GA and-857 TT were significantly associated with chronic severe hepatitis B (OR =4.176; 95% CI 2.416-7.216; OR =6.09; 95% CI 2.652-14.001). The serum TNF-α level of patients with chronic severe hepatitis B was significantly higher than that of patients with chronic hepatitis, and the difference was statistically significant between the two groups (t =3.951, P =0.000). Conclusion TNF-α-308 GA and-857 TT genotype carriers may increase the risk of chronic severe hepatitis B after HBV infection. The gene polymorphism of TNF-α gene promoter region may be closely related to the occurrence of chronic severe hepatitis B in Han population in Beijing.

Objective to investigate the preventive effect of intraperitoneal injection of schistosomiasis eggs on trinitrobenzenesulfonic acid (TNBS) enteritis in mice and the effect on the expression of toll-like receptor 4 (TLR4) in intestinal epithelium. Methods 40 BALB/c mice were randomly divided into normal control group (10 mice); TNBS enteritis group (20 rats); The mortality rate, intestinal mucosal pathological manifestations, plasma tumor necrosis factor (TNF) -α content and the expression of TLR4 in colon were evaluated by immunohistochemistry and reverse transcription (RT) -PCR. Results the mortality rate of the mice in the schistosoma egg group was lower than that in the TNBS enteritis group (20% vs 70%, P<0.05), and the intestinal inflammation was milder than that in the TNBS enteritis group (Ameho standard score 1.4±0.5 vs 4.2±0.6, P<0.01). The expression of TLR4 in colon (TLR4/β-actin absorbance ratio): TNBS enteritis group>Schistosoma egg group>normal control group (0.762±0.054 vs 0.325±0.029 vs 0.237±0.021, all P<0.01). Conclusion Intraperitoneal injection of schistosoma eggs can prevent TNBS enteritis in mice, which may be related to the down-regulation of TLR4 expression in colon.

Objective to investigate the effects of TNF-α on the expression of peroxisome proliferator activated receptor-β (PPAR β) and transcriptional activity in HaCaT keratinocytes induced by tumor necrosis factor-α (TNF-α). Methods The morphological changes of apoptotic cells were observed by Hoechst 33258 staining, and the percentage of apoptotic cells was calculated. The activity of Caspase-3 was analyzed by Caspase quantitative detection kit, and the expression of PPAR β mRNA and protein induced by TNF-α was observed by RT-PCR and Western blot. The changes of DNA binding and transcriptional activity of PPAR β mediated by TNF-α were analyzed by gel migration and retention assay and luciferase reporter gene. Results After treatment of HaCaT keratinocytes with 5, 10 and 20 ng/ml TNF-α for 24 h, Hoechst 33258 staining showed that the apoptotic cells increased with the increase of TNF-α dose, and the percentage of apoptotic cell nuclei were 12%4±3%, 32%4±4% and 57%4±4%, respectively After treating HaCaT keratinocytes with TNF-α (10, 20 ng/ml) for different time, the activity of Caspase-3 increased (P<0.01). Western blot showed that the expression of PPAR β protein in HaCaT keratinocytes increased significantly after treating with 10 ng/ml TNF-α for 12 and 24 h, and the expression of PPAR β protein in HaCaT keratinocytes increased after treating with different concentrations of TNF-α for 24 h. The expression of PPAR β mRNA was enhanced by RT-PCR after treatment with TNF-α (10, 20 ng/ml) for 3, 6 h, indicating that TNF-α can induce the expression of PPAR β. Furthermore, gel migration and retention experiments and luciferase reporter gene analysis showed that TNF-α can enhance the DNA binding and transcription activity of PPAR β. Conclusion TNF-α can enhance the expression of PPAR β mRNA and protein level, as well as the DNA binding and transcription activity in TNF-α-mediated apoptosis of HaCaT keratinocytes.

Objective to observe the inhibitory effect of small interfering RNA (siRNA) targeting mouse tumor necrosis factor α (TNF-α) gene on the expression of TNF-α in mouse macrophage line RAW264.7 in vitro.Methods Three siRNA sequences (siRNA1~3) designed for different sites of TNF-α mRNA and one fluorescent labeled BLOCK-ITTM fluorescent Oligo (modified fluorescent labeled dsRNA, siRNA4) were transfected into mouse macrophage line RAW264.7 by liposome encapsulation, and one siRNA without any target gene was set up as negative control (siRNA4). The transfection efficiency of siRNA was observed under fluorescence microscope; The inhibitory effects of siRNA on the expression of TNF-α mRNA and protein were detected by real-time fluorescence quantitative PCR and enzyme-linked immunosorbent assay (ELISA), respectively. Results the expression of TNF-α mRNA and the amount of TNF-α synthesized and secreted in macrophages increased at 6 h after endotoxin stimulation, and reached the peak at 9~12 h. The transfection efficiency of siRNA was 72% ~80% by fluorescence-labeled Oligo. After transfection of siRNA1~4, the expression of TNF-α mRNA (0.158±0.030, 0.114±0.028) and TNF-α protein stimulated by endotoxin in siRNA2 and 3 was significantly lower than that in the non-transfected group [TNF-α mRNA 0.294±0.147, protein (2104±32) pg/ml, both P<0.05], and the inhibition rate of siRNA3 was very significant, reaching 61.2% (P<0.01). The negative control siRNA4 had no effect on the expression of cellular genes and proteins

Objective to observe the inhibitory effect of bone marrow mesenchymal stem cells (MSC) on the proliferation of T lymphocytes by using the membrane surface molecules CD25 and CD69 expressed early after activation of T lymphocytes.Methods Rat MSC were expanded and identified in vitro.Rat T lymphocytes were purified by nylon cotton column.MSC and T lymphocytes were co-cultured, and the expression of CD25 and CD69 on the membrane surface of T lymphocytes was measured after stimulation with Phytohaemagglutinin (PHA). The concentrations of transforming growth factor (TGF) -β 1 and IL-10 in the supernatant of MSC culture were determined by ELISA.Results ① When more than 10,000 MSC and T lymphocytes were co-cultured, the expression of CD25 was significantly downregulated, and the downregulation was quantity-dependent. 100 MSC had no effect on the expression of CD25 in T lymphocytes; ②The down-regulation of CD25 in T lymphocytes by MSC could last for more than 96 h; ③The down-regulation of T lymphocyte CD25 by MSC is intermediate to its own secreted soluble protein; ④TGF-β 1 and IL-10 are not the reasons for the down-regulation of CD25 in T lymphocytes mediated by MSC. Conclusion MSC can regulate the immunity of T lymphocytes stimulated by PHA, and its application prospect in transplantation immunity is very bright.

Objective to investigate the effect of immunosuppressive therapy on the prevention and treatment of tissue calcification in donors of allograft with valve (AVH). Methods A modified rat model of AVH abdominal aorta ectopic transplantation was divided into allogenomic group and allogenomic group. The allogenomic group was AVH transplanted with cryopreservation, AVH transplanted with cyclosporine A (CsA), dendritic cell (DC) monoclonal antibody preconditioned with cryopreservation, and DC monoclonal antibody after transplantation. The same genome was the control group, the allogenomic SD rats were the donors, and the Wistar rats were the recipients; The expressions of T cell antigen receptors α, β (TCR-α, β) and CD28 in blood of rats were detected by flow cytometry at 2, 4, 8, 12 and 16 weeks after operation. The morphological changes of graft tissue were observed. The expression of CD54 in arterial wall was detected by immunohistochemical staining, and the content of calcium in AVH tissue was determined by atomic flame absorption spectrometry. Results (1) The expression levels of TCR-α, β and CD28 were (52.4±3.3) % and (43.8±6.4) % and (51.7±7.5) % and (66.3±4.4) % in the group receiving cryopreservation AVH at 2 and 4 weeks after operation; The expression of TCR-α and β in the CsA treatment group was (34.5±3.5) %, (31.6±2.6) %, and the expression level of CD28 was (41.2±1.6) %, (55.1±5.1) %; The expression levels of TCR-α and β in the DC monoclonal antibody treatment group were (31.6±2.3) % and (29.5±3.0) %; CD28 was (36.6±3.6) %, (51.8±5.6) %; The expression of TCR-α and β in the control group was (23.2±1.3) % and (21.6±2.3) %; CD28 was (30.7±1.4) % and (33.3±0.9) %. (2) The calcium content of donor tissues of allogenome increased at different times from 4 weeks after transplantation, but there was no significant difference in the calcium content of the same genome at each time point (P>0.05). Conclusion Immunosuppressive therapy can reduce immune response and delay the calcification process, and the curative effect is obvious.

Objective to investigate the transmission of erythromycin resistance in 30 strains of enterococcus isolated from children. Methods Thirty strains of enterococcus resistant to erythromycin were screened by agar dilution method recommended by the Institute of Clinical Laboratory Standardization (CLSI). The plasmid conjugation and transfer test was carried out by filter membrane conjugation method. The ermB gene and transposons Tn1545 and Tn917 were detected by PCR. Results Thirteen zygotes of 13 strains of enterococcus faecalis were obtained by plasmid conjugation and transfer test. The minimum inhibitory concentration (MIC) of erythromycin was ≥512 μ g/ml. Except one strain of enterococcus faecalis was negative, the other 12 zygotes all carried ermB gene, and the ermB gene was present in both transposons Tn1545 and Tn917. 16 strains of enterococcus faecalis and one strain of enterococcus faecalis were obtained by plasmid conjugation and transfer test. The erythromycin MIC was ≥512 μ g/ml. Except one strain of enterococcus faecalis was negative, the other 16 strains were ermB gene The erythromycin MIC of 30 zygotes of Staphylococcus aureus obtained by plasmid conjugation transfer test was ≥512 μ g/ml. Except for the original donor bacteria, 1 strain of Enterococcus faecalis and 1 strain of Enterococcus faecium, all the other 28 zygotes carried ermB gene. Among them, 23 strains carried ermB gene in both Tn1545 and Tn917, 4 strains carried ermB gene only in Tn1545 and 1 strain carried ermB gene only in Tn917. Conclusion The resistance of enterococcus to erythromycin can be transmitted between the same species, the same species and the different genera; The resistance of erythromycin is consistent with the ermB gene, which is closely related to Tn1545 and Tn917, and the ermB gene, Tn1545 and Tn917 can be transmitted between the same species, the same species and the different genera.

The patient was a 49-year-old male, who accidentally found a mass in the left scrotum for 2 months, occasionally with dull or mild pain. Physical examination showed a mass of 2 cm ×2.5 cm in the tail of the left epididymis, with smooth surface and tough texture, clear boundary with surrounding tissues and no tenderness.

Medical education is different from other subjects education, medical students can not become doctors directly after graduation, and need to go through hospital training and standardized training of residents to become a qualified resident.However, due to the restriction and influence of China's planned economy system, a relatively perfect training and admission management system for specialist doctors has not been established in the training system and approved person management system. As a result, most doctors engaged in clinical medical work lack strict and standardized specialist training, and even the doctors trained in large general hospitals have no authoritative qualification.

The patient, a 63-year-old female, went to our hospital in September 2005 because of "fever with abdominal pain for 2 days". The maximum body temperature was 38.6℃ in the past 2 days, and the abdominal pain was located in the left upper abdomen, accompanied by nausea and vomiting, without acid reflux, belching, diarrhea, frequent urination, urgency and pain in urination, without chest tightness and chest pain.

The patient was a 48-year-old male, unmarried, with polydipsia, polyphagia and polyuria for 5 years, and weight loss for 2 years. The patient was diagnosed with type 2 diabetes 5 years ago, and was treated with Chinese medicine irregularly, and blood sugar control was poor.The weight gradually decreased from 78 kg to 71 kg in recent 2 years.