Latest Issue
Volume 0 ， Issue 2 ， 2006
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HIGH VARIABILITY OF HUMAN CYTOMEGALOVIRUS UL150 OPEN READING FRAME IN LOW-PASSAGED CLINICAL ISOLATES
Abstract：Objective To investigate the polymorphism of human cytomegalovirus （HCMV） UL150 open reading frame （ORF） in low-passaged clinical isolates, and to study the relationship between the polymorphism and different pathogenesis of congenital HCMV infection. Methods PCR was performed to amplify the entire HCMV UL150 ORF region of 29 clinical isolates, which had been proven containing detectable HCMV-DNA using fluorescence quantitative PCR. PCR amplification products were sequenced directly, and the data were analyzed. Results Totally 25 among 29 isolates were amplified, and 18 isolates were sequenced successfully. HCMV UL150 ORF sequences derived from congenitally infected infants were high variability. The UL150 ORF in all 18 clinical isolates shifted backward by 8 nucleotides leading to frame-shift, and contained a single nucleotide deletion at nucleotide position 226 compared with that of Toledo strain. The nucleotide diversity was 0.1% to 6.8% and the amino acid diversity was 0.2% to 19.2% related to Toledo strain. However, the nucleotide diversity was 0.1% to 6.4% and amino acid diversity was 0.2% to 8.3% by compared with Merlin strain. Compared with Toledo, 4 new cysteine residues and 13 additional posttranslational modification sites were observed in UL150 putative proteins of clinical isolates. Moreover, the UL150 putative protein contained an additional transmembrane helix at position of 4-17 amino acid related to Toledo. Conclusion HCMV UL150 ORF and deduced amino acid sequences of clinical strains are hypervariability. No obvious linkage between the polymorphism and different pathogenesis of congenital HCMV infection is found.Keywords：human cytomegalovirus;gene;UL1504|10|1Updated：2024-04-10 - Abstract：Objective To evaluate the efficacy and safety of diacerein in patients with knee osteoarthritis （OA）. Methods A total of 223 patients satisfying the American College of Rheumatology criteria for knee OA were chosen for this 17-week, randomized, double-dummy, diclofenac sodium-controlled trial, with diacerein dosage of 100 mg/d and diclofenac sodium of 75mg/d. Efficacy and safety of both drugs were evaluated. Results Totally 106 patients in the diacerein group and 107 patients in the diclofenac group were considered qualified for the evaluation. After 12 weeks of treatment, the total effective rates of patients/physicians’ overall assessment in diacerein and diclofenac groups were 65.4%/61.6% and 61.2%/61.2%, respectively （P>0.05）. The primary efficacy parameter [visual analog scale （VAS） assessment of pain on 20 metres walking] and the secondary efficacy parameters [tenderness on palpation, Western Ontario and McMaster Universities Osteoarthritis Index （WOMAC）, and 36-item Short-Form （SF-36） Health Survey] significantly improved compared with baseline in both groups （P<0.05）. In the follow-up period, there were no obvious changes in above parameters in diacerein group. However, in diclofenac group, pain on 20 metres walking, tenderness on palpation, and WOMAC became aggravated after withdrawing the drug for 4 weeks （P<0.05）. Moreover, the consumption of paracetamol was significantly lower in diacerein group than in diclofenac group during follow-up （P<0.001）. The incidences of related adverse events were 35.7% in diacerein and 45.1% in diclofenac group, respectively. Mild-to-moderate gastrointestinal disorders were the most frequent adverse events. Conclusions Diacerein is as effective as diclofenac sodium in treating patients with knee OA. Furthermore, it has better extended effect and a good safety profile. It is generally well tolerated and has no severe adverse effect.Keywords：osteoarthritis;drug evaluation;diacerein;diclofenac sodium4|50|3Updated：2024-04-10
- Abstract：Objective To evaluate the overexpression of cyclin G1 in cervical intraepithelial neoplasia （CIN） and cervical carcinoma, and the correlation between cyclin G1 and high-risk human papilloma virus （HPV） infection. Methods All of the specimens were obtained from the Department of Pathology of China-Japan Friendship Hospital from January 2000 to August 2004.We detected the expression of cyclin G1 with immunohistochemistry, HPV16/18 infection with in situ hybridization, and high-risk HPV infection with Hybrid capture system II （HC-II） in normal group （25 cases）, CIN I （48 cases）, CIN II （56 cases）, CIN III （54 cases）, and invasive cervical squamous-cell carcinoma （SCC, 31 cases）. Results The positive rates of cyclin G1 expression in CIN（77.85%）and SCC cervical tissues （87.10%） were significantly higher than normal （8.00%, P<0.01）, and the intensities of cyclin G1 expression in CIN（40.60%）and SCC cervical tissues （61.51%） were significantly higher than normal （2.72%, P<0.05）. The positive rates and intensities of cyclin G1 expression increased gradually with the grade of cervical lesions. High-risk HPV infection rates were higher in CIN and SCC than normal groups （P<0.05）. There was a positive correlation between cyclin G1 expression and high-risk HPV infection detected with HC-II （Kendall’s tau-b=0.316, 0.269, 0.352, and 0.474 in CIN I, CINII, CIN III, and SCC, respectively, P<0.05）. Conclusions Cyclin G1 is overexpressed in CIN and SCC. Cyclin G1 may be a biomarker for detecting CIN and SCC. Cyclin G1 may play an important role in the oncogenesis of CIN and SCC by high-risk HPV infection.Keywords：cyclin G1;human papilloma virus;cervical intraepithelial neoplasia;cervical squamous cell carcinoma;immunohistochemistry;in situ hybridization;Hybrid capture system II4|18|9Updated：2024-04-10
- Abstract：Objective To evaluate the impact of extracapsular spread （ECS） in ipsilateral neck metastasis on prognosis and its related factors in laryngeal cancer. Methods The study included 184 patients who underwent laryngectomy and simultaneous radical or modified radical neck dissection between January 1994 and December 1997 for laryngeal cancer. All of them had a complete 5-year follow-up. We used transparent lymph node detection and continuous slicing method on all neck dissection specimens. Kaplan-Meier model was used for survival analysis and the log-rank test was used to assess significance. Results We found pathological neck metastases in 80 patients. Among them, 26 cases （32.5%） had ECS in ipsilateral neck. ECS incidence increased with advanced pathological N （pN） stages （pN1 3.7%, pN2a 25.0%, pN2b 50.0%, and pN2c 55.6%; P=0.001）. ECS incidence also increased with number of positive nodes （1 positive node 8.6%, 2 positive nodes 33.3%, 3 and more positive nodes 66.7%; P<0.001）. Incidences of contralateral neck metastases and ipsilateral neck recurrence in patients with ECS were higher than those in patients without ECS （46.2% vs. 24.1%, P=0.046; 34.6% vs. 7.4%, P=0.002）. The 5-year survival rate of patients with ECS was significantly lower than that of patients without ECS （23.1% vs. 57.4%, P=0.013）. Conclusion ECS is an important prognostic factor in laryngeal cancer. Patients with ECS have a higher incidence of contralateral neck metastasis, so bilateral neck dissection should be selected.Keywords：laryngeal neoplasms;lymphatic metastasis;extracapsular spread;prognosis4|9|3Updated：2024-04-10
- Abstract：Objective To investigate the relationship between glycosylated hemoglobin A1c （HbA1c） and blood glucose levels of eight different points throughout the day in well-glycemic-controlled medical nutrition therapy （MNT） alone type 2 diabetic patients. Methods Data were collected as capillary blood glucose value of eight different sample points among sixteen observing days in thirty MNT alone type 2 diabetic patients. The correlation between HbA1c and capillary blood glucose value was evaluated by Pearson’s correlation method. Results The r-values between HbA1c and capillary blood glucose of 3:00, 6:00, and bedtime （22:00-23:00） were 0.81, 0.79, and 0.78, respectively（P<0.001）. The best correlation was found between the mean value of 8-point blood glucose value throughout the day and HbA1c （r=0.84, P<0.001）. Conclusion Fasting blood glucose and postabsorptive blood glucose have better correlations with HbA1c compared with other points in this group of well-glycemic-controlled MNT alone type 2 diabetic patients.Keywords：type 2 diabetes;medical nutrition therapy;hemoglobin A1c;blood glucose4|21|8Updated：2024-04-10
- Abstract：Objective To investigate the influence of vitamin D receptor （VDR） gene polymorphisms on susceptibility to type 1 diabetes mellitus （T1DM） in the Chinese Han population. Method One hundred and thirty-six Chinese Han people, including 54 T1DM patients and 82 unrelated healthy subjects as control were genotyped by polymerase chain reaction-restriction fragment length polymorphism for three restriction sites in the VDR gene, which were ApaI, TaqI, and BamI. Results The frequency of B allele of BsmI site in VDR gene was significantly higher in T1DM patients than in healthy subjects （P = 0.033） while no difference was found between the two groups in the distribution of ApaI and TaqI polymorphisms. Conclusion The BsmI polymorphism of VDR gene may be associated with the susceptibility to T1DM in the Chinese Han population of Beijing.Keywords：type 1 diabetes mellitus;vitamin D receptor;gene polymorphism4|40|11Updated：2024-04-10
- Abstract：Objective To investigate the role of the burden of abnormal hematopoietic clone in the development of myelodysplastic syndromes （MDS）. Methods The ratio of the bone marrow cells with abnormal chromosomes to the total counted bone marrow cells was regarded as the index of MDS clone burden. The disease severity related parameters including white blood cell count, hemoglobin, platelet count, lactate dehydrogenase level, bone marrow blast, myeloid differentiation index, micromegakaryocyte, transfusion, interleukin-2, tumor necrosis factor （TNF）, CD4+ and CD8+ T cells of MDS patients were assayed, and the correlations between those parameters and MDS clone burden were also analyzed. Results The clone burden of MDS patients was 67.4%±36.2%. MDS clone burden positively correlated with bone marrow blasts （r = 0.483, P<0.05）, negatively with hemoglobin level （r=-0.445, P<0.05）. The number of blasts, hemoglobin, and erythrocytes in high clone burden （>50%） and low clone burden （≤50%） groups were 7.78%±5.51% and 3.45%±3.34%, 56.06±14.28 g/L and 76.40±24.44 g/L, （1.82±0.48）×10 12 /L and （2.32±0.66）×10 12 /L, respectively （all P<0.05）. CD4+ T lymphocytes of MDS patients and normal controls were （0.274±0.719）×10 9 /L and （0.455±0.206）×10 9 /L, respectively （P<0.05）. CD8+ T lymphocytes of MDS patients and normal controls were （0.240±0.150）×10 9 /L and （0.305±0.145）×10 9 /L, respectively. The serum level of interleukin-2 of MDS patients （6.29±3.58 ng/mL） was significantly higher than normal control （3.11±1.40 ng/mL, P<0.05）. The serum level of TNF of MDS patients and normal control group were 2.42±1.79 ng/mL and 1.68±0.69 ng/mL, respectively. The ratio of CD4 to CD8 was higher in high clone burden MDS patients （1.90±0.52） than that in low clone burden patients （0.97±0.44, P<0.05）. Conclusion The quantitive clonal karyotype abnormalities and deficient T cell immunity are important parameters for evaluating MDS severity and predicting its progression.Keywords：myelodysplastic syndromes;karyotype;T lymphocyte4|9|2Updated：2024-04-10
- Abstract：Objective To investigate the clinical pathological feature and diagnostic criteria of tongue amyloidosis（AL）. Methods During 1992 to 2005, 25 patients pathologically diagnosed as tongue AL in our hospital were reviewed retrospectively, and all of them had no enlarged tongue. Haematoxylin and eosin （HE） and immunohistochemical staining were used to detect the amyloid deposition on the tongue. Results Totally 84% （21/25） patients had symptoms of xerostomia and taste-blindness, 44% （11/25） patients complained of activity limitation of tongue. Macroscopic observation showed mucosa pallescence, punctuate hemorrhage, red grain particles, and ulcers on the tongue. HE staining indicated amyloid depositions in basement membrane, muscle cell, vessel wall, and nerve fiber. Immunohistochemical study demonstrated kappa light-chain deposition in 64% （16/25） cases, and lambda light-chain deposition in 36% （9/25） cases. They presented in vessel wall, nerve fiber, and muscle cells. Conclusion The biopsy is an important means for the diagnosis of early tongue AL, and the wide variety of amyloid light chain is helpful to differential diagnosis.Keywords：tongue;amyloidosis;immunohistochemistry;clinical4|10|1Updated：2024-04-10
- Abstract：Objective To investigate the effect of interleukin-6 （IL-6） on the apoptosis of annulus fibrosus （AF） cell induced by interleukin-1β （IL-1β）. Methods Cultured AF cells were divided into 6 groups and treated with no drug, 10 ng/mL IL-6,10 ng/mL IL-1β,10 ng/mL IL-1β and Z-VAD-FMK （a caspase-9 inhibitor）,10 ng/mL IL-1β and 10 ng/mL IL-6,10 ng/mL IL-1β and 100 ng/mL IL-6, respectively. After three days of culture, the apoptosis rate, the positive rates of caspase-3, -8, and -9 of AF cells were detected with flow cytometry. Results The apoptosis rates of cells in group 1 to 6 were 2.67%±1.08%, 2.71%±0.53%, 20.37%±1.57%, 11.34%±0.67%, 18.17%±0.74%, and 9.42%±1.08%, respectively. There was no significant difference between group 1 and 2, while the apoptosis rates of group 4, 5, and 6 were significantly lower than group 3 （P=0.001,P=0.172, and P=0.001, respectively）. Positive rates of caspase-3 in group 5 （12.35%±0.64%） and 6 （9.26%±0.36%） were significantly lower than group 3 （17.14%±0.72%; P=0.001 and P<0.001, respectively）. And positive rates of caspase-9 in group 5 （15.13%±1.45%） and 6 （10.17%±2.50%） were significantly lower than group 3 （19.4%±0.98%; P=0.014 and P=0.004, respectively）. But there was not obvious change of caspase-8 activity after IL-6 was added. Conclusion IL-6 is capable of protecting AF cells from IL-1β induced apoptosis in vitro. Mechanism of the protection is related with the inhibition of caspase-3 and -9 activities.Keywords：intervertebral disc;apoptosis;interleukin-6;interleukin-1β4|9|4Updated：2024-04-10
- Abstract：Objective To study the effects of constantly slow intravenous arsenic trioxide （As 2 O 3 ） infusion regimen on decreasing leukocytosis in vivo and in vitro. Methods Three kinds of leukemia cells, NB4, K562, and acute promyelocytic leukemia （APL） cells, were cultured in the media with constant concentration and varying concentrations of As 2 O 3 respectively for 24 hours. Seventy-five patients were enrolled in two groups randomly. In trial group, 37 patients received continuously slow intravenous As 2 O 3 infusion regimen for 24 hours with an infusion rate of 8 drips per minute and total infusion duration of about 18-21 hours daily. In control group, 38 patients received routine regimen with an infusion rate of 45-55 drips per minute and total infusion duration of about 2-3 hours daily for 24 hours. The daily As 2 O 3 dosage was 0.16 mg/kg. The intracellular arsenic concentration was measured by atomic fluorescence assay. The apoptosis rate of cells, CD33 - CD11b + cells, and CD33 + CD11b - cells were monitored by flow cytometry. Results The apoptosis rates of NB4, K562, and APL leukemia cells in the media with constant As 2 O 3 concentration were 56.6%±2.4%, 27.6%±3.1%, and 52.2%±2.8%, respectively, which were significantly higher than those with changing As 2 O 3 concentration （23.2%±2.1%, 11.0%±2.5%, and 21.0%±2.5%, respectively, P<0.01）. The apoptosis rates of APL, M2 type acute myeloid leukemia （AML-M2）, and chronic myeloid leukemia （CML） patients in the trial group （28.5%±1.9%, 9.5%±0.6%, and 12.5%±1.8%） were also significantly higher than those in control group （8.5%±2.2%, 2.9%±0.8%, and 4.5%±1.2%; P<0.05）. The ratios of CD33 + CD11b - and CD33 - CD11b + cells in control group were significantly higher than those in trial group. Conclusion The continuously slow intravenous As 2 O 3 infusion regimen can obtain high efficiency of apoptosis and low differentiation proportion, relieve leukocytosis, and gain maximal therapeutic benefit.Keywords：arsenical;infusion regimen;leukocytosis4|35|3Updated：2024-04-10
- Abstract：Objective To construct an expression vector of small interfering RNA （siRNA） against survivin and observe its effects on survivin expression and proliferation of human pancreatic cancer cell line PC-2 and breast cancer cell line MCF-7. Methods Constructed an expression vector of siRNA against survivin and transfected it into PC-2 and MCF-7 cells using lipofectamine TM 2000.The expression of survivin was detected by semi-quantitive RT-PCR and immunohistochemistry, and its effects on proliferation of PC-2 and MCF-7 cells were detected by MTT assay. Results The introduction of sequence-specific siRNA could efficiently suppress survivin expression at both mRNA and protein levels in the two cancer cell lines. In PC-2 cells, the expression inhibition rates were 81.25% at mRNA level and 74.24% at protein level. In MCF-7 cells, the expression inhibition rates were 64.91% at mRNA level and 79.72% at protein level. The proliferation of PC-2 and MCF-7 cells was also suppressed, and 24 and 48 hours after the cells were reseeded, the proliferation inhibition rates of PC-2 cells were 28.00% and 33.38%, and that of MCF-7 cells were 31.58% and 33.02%, respectively. Conclusions The expression vector of siRNA against survivin can block survivin expression in PC-2 and MCF-7 cells efficiently and specifically. Down regulation of survivin expression can suppress proliferation of PC-2 and MCF-7 cells. Survivin RNAi may have potential value in gene therapy of human cancers.Keywords：RNA interference;small interfering RNA;Survivin;pancreatic cancer;breast cancer4|23|12Updated：2024-04-10
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GENETIC ENGINEERING NEURAL STEM CELL MODIFIED BY LENTIVIRUS FOR REPAIR OF SPINAL CORD INJURY IN RATS
Abstract：Objective To explore the feasibility for therapy of spinal cord injury （SCI） by genetic engineering neural stem cell （NSC） modified by lentiviral vector. Methods Following the construction of the genetic engineering NSC modified by lentivirus to secrete both neurotrophic factor-3 （NT-3） and green fluorescence protein （GFP）, hemisection of spinal cord at the level of T 10 was performed in 56 adult Wistar rats that were randomly divided into 4 groups （n=14）, namely 3 therapeutic groups and 1 control group. The therapeutic groups were dealed with NSC, genetic engineering NSC, and concentrated lentiviral supernatant which carries both GFP and NT-3, respectively. Then used fluorescence microscope to detect the transgenic expression in vitro and in vivo, migration of the grafted cells in vivo, and used the Basso, Beattie, and Bresnahan （BBB） open-field locomotor test to assess the recovery of function. Results The transplanted cells could survive for long time in vivo and migrate for long distance. The stable transgenic expression could be detected in vivo. The hindlimb function of the injured rats in 3 therapeutic groups, especially those dealed with genetic engineering NSC, improved obviously. Conclusion It is feasible to combine NSC with lentivirus for the repair of SCI. NSC modified by lentivirus to deliver NT-3, acting as a source of neurotrophic factors and function cell in vivo, has the potential to participate in spinal cord repair.Keywords：lentivirus;spinal cord injury;neural stem cell;neurotrophic factor-34|105|14Updated：2024-04-10 - Abstract：Objective To compare the effectiveness of immunofluorescence and polymerase chain reaction （PCR） in detecting human papilloma virus （HPV） in condyloma acuminata （CA）. Methods HPVs in CA tissues from 60 patients were detected by immunofluorescence and PCR, respectively. Different subtypes of HPVs were also identified with restriction fragment length polymorphism （RFLP）. Results The positive detective rates of immunofluorescence and PCR were 56.67% （34/60） and 96.67% （58/60）, respectively （P<0.01）. RFLP results showed HPV6 and HPV11 were the main subtypes in the detected virus, which accounted for 98.28%. Conclusion The sensibility of PCR is superior to that of immunofluorescence.Keywords：condyloma acuminata;human papilloma virus;polymerase chain reaction;immunofluorescence4|23|0Updated：2024-04-10
- Abstract：Objective To investigate the effect of glycosylation at Asn302 of pro-urokinase （pro-UK） on the stability in culture supernatant. Methods Nonglycosylated pro-UK was constructed by site-directed mutagenesis of Asn302 to Ala302.The pro-UK mutant and native pro-UK were transfected into dhfr - -CHO cells, and serum-free culture supernatant was harvested and incubated at 4℃ and 37℃, respectively. The pro-UK activity in culture supernatant was measured by the optical density （OD） increase with time （12 hours） at 405 nm. Without thermolysin activation, the percentage of single chain pro-UK was measured. Results After 48 hours of incubation at 4℃, the activities of pro-UK mutant and native pro-UK decreased 3.7% and 2.9% respectively, and at 37℃ decreased 37.9% and 23.5%, respectively. The total activity of native pro-UK was significantly higher than that of nonglycosylated mutant at 37℃. The single-chain percentage of native pro-UK was higher than that of nonglycosylated mutant at both 4℃ and 37℃. Conclusion Higher temperature increases the proteolysis of pro-UK. The glycosylation site on Asn302 is beneficial to pro-UK stability in culture supernatant.Keywords：stability;pro-urokinase;glycosylation4|14|1Updated：2024-04-10
- Abstract：Objective To study the therapeutical effects of crossing anastomosis of nerve on the peripheral and central nerve injuries. Methods Twelve kinds of central and peripheral nerve disorders and their complications were treated with 11 kinds of crossing anastomosis of nerve bundles near the innervated organs. After nerve injury and repair, somatosensory evoked potentials （SEPs） and horseradish peroxidase （HRP） retrograde tracing studies were used to investigate the rabbit’s nerve function and morphology. Results The ulcers of all patients healed. Sensation, voluntary movement, and joint function recovered. Four weeks after the anastomosis of distal stump of radialis superficialis nerve and median nerve, pain sensation regained and SEPs appeared. HRP retrograde tracing studies demonstrated sensory nerve ending of medial nerve formed new connection with the body of neuron. Conclusion Crossing anastomosis of nerve is an effective method to treat peripheral and central nerve injuries.Keywords：anastomosis of nerve;nerve injury4|7|0Updated：2024-04-10
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