HLA-DRB1 genotyping and its relation with chronic HBV infection in patients

AIM: To study the HLA-DRB1 genotype and their relation with HBV infection in Shaanxi Han patients. METHODS: HLA-DRB1 genotyping was conducted in 108 case of chronic HBV infection and 108 health controls as well as 32 asymtomatic HBsAg carriers by using polymerase chain reaction-sequence specific primer method. All the patients, asymtomatic HBsAg carriers and health subjects were residents of Shaanxi district and belonged to Han nationality. The association between HLA-DRB1 genotype and different replication of HBV was also studied. RESULTS: DRB1*04, DRB1*09, DRB1*12 and DRB1*15 were the most common genotypes in Shaanxi Han residents with the frequency of 16.2%, 12.5%, 11.6% and 13.4%, respectively. Compared to 108 health controls, the allele frequency of HLA-DRB1*03 (10.6% of HBV patients versus 3.7% of health controls, odds ratio=3.10; P<0.05) and HLA-DRB1*07 (17.6% of HBV patients versus 9.3% of health controls, odd ratio=2.09; P<0.05) were markedly higher. The allele frequency of HLA-DRB1*15 (13.4% of HBV patients versus 6.9% of health controls, odds ratio=0.48; P<0.05) was obviously lower than than in HBV patients. CONCLUSION: HLA-DRB1*03 and HLA-DRB1*07 are closely related with susceptibility to chronic hepatitis B infection, and DRB1*15 is closely related with resistance to chronic hepatitis B infection. These finding suggest that host HLA class II gene is an important factor determining the outcome of HBV infection.     

Relationship between UGT1A1 gene polymorphisms and toxicity/efficacy of irinotecan-based chemotherapy in metastatic colorectal cancer

AIM: To investigate the correlation of UGT1A1 ^*28 and UGT1A1 ^*6 gene polymorphisms with irinotecan-associated adverse events and efficacy in the patients with metastatic colorectal cancer (mCRC) treated with irinotecan-based chemotherapy. METHODS: Analysis of UGT1A1 ^*28 and UGT1A1 ^*6 gene polymorphisms was performed in 207 gastrointestinal cancer patients admitted to our hospital from April 2010 to March 2012 by amplifying the gene fragments using PCR and direct sequencing. Fifty six cases with mCRC treated with irinotecan were chosen to observe the adverse events and efficacy during chemotherapy, and the time to progression (TTP) was also recorded. The incidence of different genotypes was compared. RESULTS: The distribution of the genotypes in 207 gastrointestinal cancer patients was as follows: UGT1A1 ^*28 wild-type (WT) genotype TA6/6 (164, 79.2%), heterozygous genotype TA6/7 (41, 19.8%), and homozygous genotype TA7/7 (2, 1.0%); UGT1A1 ^*6 WT genotype G/G (154, 74.4%), heterozygous genotype G/A (51, 24.6%), and homozygous genotype A/A (2, 1.0%). In the 56 mCRC cases, the incidence of grade 3 and 4 delayed diarrhea and neutropenia in the patients carrying UGT1A1 ^*6 (G/A and A/A) was higher than that in the WT genotype (6/6) (38.9% vs 7.9%,61.1% vs 29.0%, both P<0.05). The incidence of grade 3 and 4 thrombocytopenia in the patients carrying UGT1A1 ^*28 (TA6/7 and TA7/7) was higher than that in the WT genotype (TA6/6) (33.3% vs 2.1%, P<0.05). No significant difference of TTP and chemotherapeutic effect was observed between different genotypes. CONCLUSION: The UGT1A1 ^*6 (G/A and A/A) genotypes increase the risk of grade 3 and 4 delayed diarrhea and neutropenia, and the UGT1A1 ^*28 (TA6/7 and TA7/7) genotypes increase the risk of grade 3 and 4 thrombocytopenia in mCRC patients treated with irinotecan-based chemotherapy.     

Relationship between 936C/T polymorphism in 3’-untranslated region of vascular endothelial growth factor gene and diabetic retinopathy in China

AIM: To study the distribution of 936C/T polymorphism in 3’- untranslated region of vascular endothelial growth factor (VEGF) gene in Chinese Han population and to analyze the relationship between the polymorphism and diabetic retinopathy (DR). METHODS: Two hundred and fifty-four patients with type 2 diabetes mellitus recruited in this study were divided into NPDR group (non-proliferative diabetic retinopathy), PDR group (proliferative diabetic retinopathy) and DM (diabetes without retinopathy) group. The normal control group consisted of 120 subjects. Genotypes were identified by PCR-RFLP among all the subjects, while other clinical parameters were measured. Serum levels of VEGF were tested by the method of ELISA. RESULTS: The frequencies of both genotype CC and allele C were significantly higher in NPDR group and PDR group than those in NC group (²=9.934, ²=4.899, P<0.05 and ²=10.895, ²=5.714, P<0.05) and DM group (²=7.490, ²=4.448, P<0.05 and ²=8.333, ²=5.227, P<0.05). However, the frequencies of genotype (TT+CT) and allele T were significantly lower in NPDR group and PDR group than those in NC group (²=9.934, ²=10.895, P<0.01 and ²=4.899, ²=5.714, P<0.05) and DM group (²=7.490, ²=8.333, P<0.01 and ²=4.448, ²=5.227, P<0.05). Multivariate logistic regression analysis showed that the levels of glycohemoglobin(HbA1c), total cholesterol(TC),low-density lipoprotein cholesterol(LDL-C) and plasma VEGF presented a positive correlation with DR, respectively, and the 936C/T mutation of VEGF exhibited a negative correlation with DR (β=-1.027, OR=0.343, P<0.01, CI: 0.157-0.723). CONCLUSION: Allele 936C of VEGF may be a genetic marker susceptible to DR, while allele 936T may be a protective genetic marker of DR. The 936C/T mutation of VEGF may be a protective factor against DR.     

Regulation of LPS-induced elevation of Ca2+ intracellular level of alveolarmacrophages in chronic bronchitis by Angelica Sinensis and nifedipine

AIM: To explore regulation of lipopolysaccharide (LPS)-induced elevation of Ca²⁺ intracellular level in alveolar macrophages(AMs) from patients with chronic bronchitis by Angelica Sinensis and nifedipine.METHODS:AMs was obtained from 7 patients with chronic bronchitis and 6 normal controls by bronchoalveolar lavage and intracellular Calevel was detected after adding Angelica Sinensis, nifedipine or LPS to the supernatant of AMs loaded by Fura-2. RESULTS: In contrast with normal control group (99.65±32.21 nmol/L), intracellular Ca²⁺ level in AMs from chronic bronchitis group (189.47±23.69 nmol/L) was increased significantly in the absence of extracellular Ca²⁺ but not 1 mmol/L. Intracellular Ca²⁺ level in AMs from chronic bronchitis group were significantly increased by adding 10 μg/mL LPS to the supernatant of AMs. LPS-induced elevation of intracellular Ca²⁺ level in AMs from chronic bronchitis group was completely inhibited by Angelica Sinensis or nifedipine.CONCLUSION: Both Anelica Sinensis and nifedipine may inhibit activation of AMs from patients with chronic bronchitis by reducing LPS-induced elevation of intracellular Ca²⁺ level in AMs, suggested that these two medicines may inhibit non-specific inflammation of airways in chronic bronchitis.     

Detection and clinical significance of myeloid-derived suppressor cells in peripheral blood of patients with Parkinson disease

AIM: To detect the myeloid-derived suppressor cells (MDSCs) in peripheral blood from the patients with Parkinson disease (PD) and its clinical significance. METHODS: The patients (n=80) diagnosed PD from January 2016 to March 2017 in our hospital and 20 healthy volunteers were selected as the subjects. According to the Hoehn-Yahr staging, 80 PD patients were staged, of whom 22 were I, 24 were Ⅱ, 20 were Ⅲ, 14 were IV, and 0 was V. Peripheral blood (5 mL) samples from the patients with PD and the healthy volunteers were collected and the mononuclear cells were isolated. The levels of CD14⁺CD11b⁺ cells and CD14^-CD11b⁺ cells in the peripheral blood were detected by flow cytometry. The two populations of the cells were sorted by magnetic beads. The mRNA levels of arginase 1 (ARG1), interleukin-10 (IL-10) and cyclooxygenase 2 (COX-2) were detected by qPCR. The expression of surface membrane proteins CD14 and CD11b, and immunosuppressive factors ARG1, IL-10 and COX-2 was determined by Western blot and ELISA. RESULTS: No significant change of CD14⁺CD11b⁺ cells between the patients with PD and normal controls was observed, but the cells with CD14^-CD11b⁺ increased significantly in the patients with PD compared with the control people (P<0.05). The CD14^-CD11b⁺ cells in peripheral blood of the patients were related to the stage of Hoehn-Yahr. The CD14^-CD11b⁺ and CD14⁺CD11b⁺ cells showed high levels of IL-10 and COX-2, and the high level of ARG1 was only expressed in the CD14^-CD11b⁺ cells. The expression of ARG1 in the CD14^-CD11b⁺ population from PD patients was significantly different from that of CD14⁺CD11b⁺ population and normal subjects (P<0.05). CONCLUSION: The CD14^-CD11b⁺ cells and ARG1 expression level in peripheral blood of the PD patients can be used to evaluate the pathogenesis and staging. Immunosuppression may play an important role in the pathogenesis and development of PD.     

Expression of Foxp3+ Tregs and PD1 in gastric cancer tissues and their correlation with clinicopathological factors and prognosis

AIM: To investigate the expression of Foxp3⁺ regulatory T cells (Foxp3⁺ Tregs) and programmed death receptor 1 (PD1) in gastric cancer tissues and their association with clinicopathological factors and prognosis of the patients. The correlation between the 2 molecules was also analyzed at the same time. METHODS: The tumor sections from 111 gastric cancer patients were stained for Foxp3 and PD1 by the method of immunohistochemistry. The associations of the expression levels of these 2 molecules with clinicopathological factors involved in the disease progression and prognosis were statistically analyzed. The relationship of their expression was detected. RESULTS: Foxp3⁺ Tregs and PD1 were expressed in the gastric cancer tissues, and PD1 was expressed in the tumor infiltrating lymphocytes (TILs). The expression of Foxp3 and PD1 was correlated with lymph node metastasis, clinicopathological stage and prognosis of gastric cancer patients. The expression of these 2 determinants in the patients with lymph node metastasis and an advanced clinicopathological stage was distinctly higher (P <0.05). The patients with positive expression of the 2 indexes presented a lower overall survival rate and worse prognosis (P <0.05). A significantly positive correlation between the infiltration of Foxp3⁺ Tregs and the expression of PD1⁺ TILs was also observed (P <0.01).CONCLUSION: Foxp3⁺ Tregs and PD1⁺ TILs co-infiltrate in the gastric cancer tissues, which can be used as biological markers to predict the disease progression and prognosis.     

C*HSDGIC* from cyclization of PACAP (1-5) attenuates UVB-induced apoptosis of human retinal pigment epithelial cells

AIM: To investigate the influence of C^*HSDGIC^* (CHC), a cyclopeptide from the cyclization of with disulfide, on the proliferation and apoptosis of cultured human retinal pigment epithelial (RPE) cells induced by ultraviolet B(UVB). METHODS: The expression of PACAP type 1 (PAC1) receptor in human RPE cells was identified by Western blotting. The cells were exposed to UVB irradiation and cultured in fresh medium with or without gradient concentrations (1 nmol/L to 1 mmol/L) of CHC. The viability of the cells was determined by CCK-8 assay. The early apoptosis of the cells was detected by flow cytometry with annexin V and propidium iodide staining.The mitochondrial menbrane potential was detected by flow cytometry with JC-1 staining. RESULTS: The PAC1 receptor in human RPE cells was identified by Western blotting. The best results of CHC on the proliferation and anti-apoptosis of human RPE cells were achieved at the concentration of 100 μmol/L, which increased the viability by (34.23±3.39)% and (20.10±1.48)%, respectively. The percentage of apoptotic cells was decreased by (5.63±1.49)% with CHC treatment (100 μmol/L) after UVB irradiation,and the percentage of mitochondrium-depolarizing cells was decreased by (5.2±0.5)%. CONCLUSION: PAC1 receptor exists in human RPE cells. C^*HSDGIC^* increases the viability of RPE cells and attenuates UVB-induced apoptosis.     

紫斑牡丹花色表型数量分类研究

以190个紫斑牡丹品种为材料,利用色差仪连续两年在盛花期测量花色参数,将所测的CIELab表色空间的L*、a*、b*值转化成蒙塞尔颜色系统的H(Hue)、V(Value)、C(Chroma)值,然后根据L*、a*、b*值系统聚类分析,结合H、V、C值,比对ISCC-NBS色名表示法对每种花色进行定性描述.结果 表明:通...     

Association analysis between two SNPs in SNCA and PARK16 genes and Parkinson disease in a Han Chinese population in Liaoning area

AIM: To investigate the genotypic frequency of rs3857059 in SNCA gene and rs16856139 in PARK16 gene for determining the potential genetic risk factors of Parkinson disease (PD) in a Han Chinese population in Liaoning area of China. METHODS: The genomic DNA from 213 PD patients and 214 matched controls was amplified in the multiplex PCR system and subsequently genotyped by digestion with endonuclease Pvu II. Genetic parameter and association studies were carried out with SPSS 13.0 and PLINK 1.07 software. RESULTS: We accurately detected all genotypes in the 2 loci with PCR-restriction fragment length polymorphism (RFLP) techniques. The gene frequency of G allele in the rs3857059 locus was higher in PD group than that in control group with statistical significance (χ²= 7.592,P<0.01, OR=0.677, 95% CI=0.517~0.888). The T allele frequency in the rs16856139 locus was lower in PD group than that in control group and statistical result revealed a significant difference (χ²=11.511, P<0.01, OR=0.390, 95% CI=0.227~0.669). CONCLUSION: The 2 SNPs investigated in SNCA and PARK16 genes are likely to play roles as common risk factors for PD disease in the Han Chinese population.     

Metallothionein protects rat hepatic nuclear nucleoside triphosphatase from hydroxyl radical-induced suppression

AIM:Metallothioneins (MTs) are cysteine-rich metal-binding proteins that exert cytoprotection during metal exposure and oxidative stress. The present study was designed to investigate whether MT can directly protect NTPase on nuclear envelope from damage induced by hydroxyl radical.METHODS:Isolated hepatic nuclei from rat liver were exposed to Fe²⁺/H₂O₂ with or without MT, and the NTPase activity on nuclei was assayed using ATP and GTP as substrate, respectively.RESULTS:Incubation of rat hepatic nuclei with the Fe²⁺/H₂O₂ (in μmol·L^-1/μmol·L^-1 : 0.1/0.5, 0.5/2.5, 1/5, 5/25) resulted in a concentration-dependent decrease in nuclear NTPase activities (P＜0.01). Incubation of hepatic nuclei with different concentrations of MT (10^-9-10^-4mol·L^-1)and Fe²⁺/H₂O₂ (1 μmol·L^-1/5 μmol·L^-1) for 10 min, nuclear NTPase activities were increased in a MT concentration-dependent fashion as compared with that of incubation with Fe²⁺/H₂O₂(1 μmol·L^-1/5 μmol·L^-1) alone. When MT was at 10^-4 mol·L^-1, TNPase activities reversed to (102±10) nmol·mg^-1 protein·min^-1(for ATP as substrated) and (131±12) μmol·g^-1 protein·min^-1(for GTP as substrate), which had no significant defferences from that of the controls (112±8 and 142±10 μmol·g^-1 protein·min^-1, respectively) (P＞0.05). In addition, incubation of hepatic nuclei with only MT had no effect on nuclear NTPase activity. CONCLUSION:These data demonstrate that hydroxyl radical generated from Fe²⁺/H₂O₂ might attack nuclear NTPase. MT antagonistically reduces toxicity of Fe²⁺/H₂O₂ system to the NTPase.     

Changes of myocardial nuclear membrane Ca2+-ATPase function in ischemia/reperfusion injury

AIM: The changes of myocardial nuclear membrane Ca²⁺ -ATPase function was investigated in ischemia/reperfusion injury. METHODS: The model of myocardial ischemia/reperfusion injury was established in rats. Myocardial nuclei were purified with sucrose density centrifugation, the activity of Ca²⁺ -ATPase was measured and calcium uptake was assayed with [⁴⁵ Ca²⁺ ] . RESULTS: Plasma levels of malondialdehyde (MDA) and free fatty acid (FFA) in myocardial ischemia/reperfusion injury increased significantly( P<0.01 vs control). Ca²⁺ -ATPase activity and [⁴⁵ Ca²⁺ ] uptake was lower than normal at below 10 μmol/L, while higher at 50 μmol/L. CONCLUSION:These data indicate dysfunction of nuclear menbrane calcium pump and [⁴⁵ Ca²⁺ ] uptake function in myocardial ischemia/reperfusion injury.     

Protective effect of sodium selenite on human keratinocytes from ultraviolet-B irradiation

AIM: To investigate the protective effect of sodium selenite (Na₂SeO₃) on human keratinocytes under ultraviolet-B (UVB) irradiation. METHODS: The cultured HaCaT cells were divided into 4 groups: (1) normal control group; (2) Na₂SeO₃ group: pretreated with Na₂SeO₃ at doses of 10 nmol/L, 50 nmol/L, 100 nmol/L, 200 nmol/L and 1 μmol/L for 24 h; (3) UVB group: irradiated with UVB at doses of 300, 600 and 900 J/m² ; (4) Na₂SeO₃+UVB group: after pretreated with Na₂SeO₃ for 24 h, irradiated with UVB at doses of 300, 600 and 900 J/m² . The cell proliferation was detected by MTT assay. The apoptotic rates of HaCaT cells treated with UVB at dose of 300 J/m² were assessed by flow cytometry. RESULTS: Compared with normal control group, the cell proliferation activity in UVB group decreased significantly (P<0.05). The cell activity was inversely correlated with the irradiation intensity. No significant difference of the cell activity between Na₂SeO₃ group and normal control group was observed. The cell proliferation in Na₂SeO₃+UVB group was higher than that in UVB group significantly (P<0.05). Na₂SeO₃ at concentration of 100 nmol/L showed the strongest activity to promote cell proliferation. After 300 J/m² UVB irradiation, the apoptotic rate in Na₂SeO₃+UVB group decreased significantly (P<0.05) compared with UVB group. The inhibitory effect of Na₂SeO₃ at concentration of 100 nmol/L on apoptosis was the strongest.CONCLUSION: The damage of human keratinocytes by UVB irradiation is in a dose-dependent manner. The photoprotection performance of Na₂SeO₃ reduces the damage of human keratinocytes induced by UVB irradiation.     

Detection and clinical significance of circulating tumor cells in peripheral blood of nasopharyngeal cancer patients with modified immunomagnetic enrichment and fluorescent immunocytochemistry

AIM: To investigate the clinical significance of the new method of modified immunomagnetic enrichment of tumor cells in combination with fluorescent immunocytochemistry to detect circulating tumor cells (CTCs) in peripheral blood samples of patients with nasopharyngeal cancer (NPC). METHODS: Peripheral blood samples were obtained from 76 histology-proven patients with NPC before the initial therapy. After isolation of the mononuclear cells, the CTCs expressing cytokeratin (CK)8/18 in the blood samples were detected by the method of immunomagnetic enrichment in combination with fluorescent immunocytochemistry. The magnetic beads covalent binding with epithelial cell adhesion molecule (EpCAM) antibody were used to enrich the tumor cells which expressed EpCAM. After median following-up for 25 months, the effects of CTCs and other prognostic factors on patient prognosis were thoroughly investigated. RESULTS: None of the positive CK8/18 cells was detected in 20 normal blood samples. The CTCs were detected in 82.9% of the patients (P<0.01). Relapse patients had significantly higher number of median CK8/18⁺ CTCs than the patients without relapse (P<0.01). No association of viral capsid antigen (VCA)-IgA (P>0.05) was observed between the patients with and without relapse. Relapse-free survival rates were lower when the number of peripheral blood CK8/18⁺ CTCs was more than 3. The 2-year relapse-free survival rates were 100%, 100%, 100%, 94.1%, 71.4%, 53.3% and 44.4%(P<0.01) when the numbers of peripheral blood CK8/18⁺ CTCs were 0, 1, 2, 3, 4, 5, 6 before treatment, respectively. Overall survival rates were lower when the number of peripheral blood CK8/18⁺ CTCs was more than 5, but the difference was not significant. The 2-year overall survival rates were 100%, 100%, 100%, 100%, 100%, 100%, 80% and 77.8% (P>0.05) when the numbers of peripheral blood CK8/18⁺ CTCs were 0, 1, 2, 3, 4, 5, 6 before treatment,respectively. The VCA-IgA titer could not predict survivals. Cox multivariate analysis also showed the same results. CONCLUSION: Peripheral blood CK8/18⁺ CTCs are a prognostic factor for initial treatment of NPC.     

Clinicopathological significance of pEZRThr567 expression in lung squamous carcinoma

AIM: To investigate the clinicopathological significance of the protein expression of phosphorylated ezrin at threonine 567 (pEZR^Thr567) in lung squamous cell carcinoma, adjacent tissues and normal tissues. METHODS: pEZR^Thr567 protein was detected in lung squamous carcinoma, adjacent and normal tissues by the method of immunohistochemistry. The correlation of pEZR^Thr567 expression with clinicopathological parameters of lung squamous carcinomas was also analyzed. The localization of pEZR^Thr567 was detected by immunofluorescence staining in lung squamous cell line EBC-1. RESULTS: The protein expression of pEZR^Thr567 in lung squamous carcinoma was significantly higher than that in the adjacent and normal lung tissues (P<0.01). pEZR^Thr567 mainly localized on the cell membrane, and its over-expression signi-ficantly correlated with the differentiation, clinical stage and lymph node metastasis in lung squamous carcinoma. CONCLUSION: pEZR^Thr567 may be an effective biomarker for prediction of malignant potential and poor prognosis of lung cancer.     

Study on the binding capacity of IgA1 from patients with IgA nephropathy to human mesangial cells

AIM: To examine and compare the ability of serum IgA 1, from both the patients with IgA nephropathy(IgAN) and the healthy control, to bind to human mesangial cells(HMC). METHODS: Serum IgA was isolated with jacalin column, heated to aggregated form(IgA₁) and labeled with [¹²⁵I]. Binding capacity of IgA₁ to primary HMC was evaluated by radioligant binding assay, specificity of binding was determined by competitive inhibition, and relative affinities was compared by cross competitive inhibition. RESULTS: Both IgA₁ from normal control and patients with IgAN bound to MC in a dose-dependent, saturatable manner, but the binding of IgA₁ from patients was saturated at approximately 200 pmol while that from healthy was at 400 pmol. The Scatchard analysis revealed a Kd of(8.9±2.1)×10^-8 mol/L for patient' s IgA₁ versus(4.3±1.2)×10^-7mol/L for normal IgA₁(P<0.05). Competition inhibition showed that both serum albumin and IgG could not block the binding of IgA₁ to HMC while mIgA 1 could partially block that. Cross competition inhibition demonstrated that patient' s IgA₁ blocked normal IgA₁ binding significantly(P<0.01), however, normal IgA₁ was unable to inhibit the binding of patient' s IgA₁. CONCLUSIONS: ①There are IgA binding proteins or receptors on mesangial cells. ②IgA₁ from patients with IgAN has a higher binding capacity than that from healthy.     

Establishment of method for PD-1 immunophenotype detection in TCR Vβ repertoire of CD3+, CD4+ and CD8+ T-cell subsets

AIM:To establish the method for detecting the immunophenotype of immunosuppressive receptor programmed cell death protein 1 (PD-1) in T-cell receptor (TCR) Vβ repertoire of CD3⁺, CD4⁺ and CD8⁺ T-cell subsets, therefore to evaluate the distribution of PD-1 in T-cell repertoire from human peripheral blood (PB). METHODS:The PB samples from 10 cases of healthy individuals (HI) were collected. Using multi-colored fluorescence flow cytometry, the distribution frequency of PD-1 in TCR Vβ repertoire was detected with a wide panel of anti-CD45, anti-CD3, anti-CD4, anti-CD8, anti-PD-1 and 24 anti-TCR Vβ repertoire (IOTest^® Beta Mark TCR Vβ Repertoire Kit, containing 8 tubes which labeled A~H, each tube is a composite antibody of FITC and PE coupling, each cocktail contains antibodies direc-ted to 3 different Vβ subfamilies) monoclonal antibodies. RESULTS:The total number of the 24 TCR Vβ repertoire detected in CD3⁺, CD3⁺CD4⁺ and CD3⁺CD8⁺ T cells from 10 cases of HI was consistent with the Quick Reference Card data provided by the kit. The preliminary results showed that the frequency of Vβ usage in CD3⁺, CD4⁺ and CD8⁺ T cells was different. High usage of Vβ2, Vβ3, Vβ8, Vβ9, Vβ5.1, Vβ13.1 and Vβ13.2 was found in CD3⁺ T cells, while high usage of Vβ2, Vβ3, Vβ8, Vβ5.1, Vβ9 and Vβ13.1 in CD3⁺CD4⁺ T cells, and high usage of Vβ1, Vβ2, Vβ3, Vβ9, Vβ13.1 and Vβ13.2 in CD3⁺CD8⁺ T cells were also observed. Further analysis showed that the expression of PD-1 was detected in all 24 TCR Vβ subfamilies of CD3⁺, CD3⁺CD4⁺ and CD3⁺CD8⁺ T cells. The higher frequency of PD-1⁺ T cells was CD4⁺Vβ5.2⁺ T cells, whereas the higher frequency of PD1⁺ T cells in CD8⁺Vβ11⁺ and CD8⁺Vβ13.6⁺ T cells was detected. CONCLUSION:The method for detection of the immunosuppressive receptor PD-1 in TCR Vβ repertoire of T-cell subsets is successfully established, which provides a new method for further analysis of immunosuppressive characteristics of TCR Vβ repertoire in the patients with leukemia.     

Effects of non-ventilated lung with nitrous oxide on intrapulmonary oxygenation and lactic acid level in arterial blood during one lung anesthesia

AIM: To investigate the effects of non-ventilated lung with N₂O on systemic oxygenation and lactic acid level in arterial blood during one lung anesthesia. METHODS: Twenty-two patients, ASA Ⅰ-Ⅲ, scheduled for selective pulmonary surgery, were randomly divided into two groups: control group (group A, n=11) and observation group (group B, n=11). Group A: the non-ventilated lung was kept open to the air; group B: N₂O 2 cmH₂O through CPAP system was insufflated into the non-ventilated lung during one lung ventilation. The anesthesia was induced with intravenous midazolam (0.05 mg·kg^-1), propofol (0.5-1.0 mg·kg^-1), fentanyl (4 μg·kg^-1), and vecuronium (0.1 mg·kg^-1) and was maintained with inhaling isoflurane. Blood gas analysis and lactic acid was recorded 20 min after two-lung ventilation (TLV) in the supine position, 20 min after one-lung ventilation (OLV) in the supine position, 20 min and 40 min after OLV in the lateral position and at the end of operation and the shunt fraction was calculated. RESULTS: PaO₂ in group B was significantly higher than that in group A (P＜0.05). Qs/Qt in group B was significantly lower than that in group A (P＜0.05), and lactic acid level in group B was significantly lower than that in group A during OLV. CONCLUSION: Non-ventilated lung with N₂O (2 cmH₂O) improves systemic oxygenation, reduces intrapulmonary shunt and prevents hypoxemia during OLV.     

Role of endogenous nitric oxide in apoptosis of alveolar epithelial cells in the development of pulmonary fibrosis in rats

AIM: To study the role of high level of endogenous nitric oxide (NO) in apoptosis of alveolar epithelial cells in the development of pulmonary fibrosis in rats. METHODS: The content of nitrite/nitrate (NO₂^-/NO₃^-) in out-flowing pulmonary blood (OPB) was assayed by nitric acid reduction method. The apoptosis of alveolar epithelial cells was observed by TdT-mediated dUTP nick-end labeling (TUNEL) and electron microscopy, respectively. The above indices were observed on the day 14 and the day 30 after intratracheal administration of BLMA₅ alone or along with blockade of iNOS by aminoguanidine (AG) in rats. RESULTS: (1) Both the content of NO₂^-/NO₃^- in OPB and the number of apoptotic alveolar epithelial cells in lung were increased in BLMA₅ 14 d group, compared with normal control group and BLMA₅ 30 d group, respectively (P<0.05). The high level of NO₂^-/NO₃^- in OPB and the apoptosis of alveolar epithelial cells were ameliorated by AG. CONCLUSION: The apoptosis of alveolar epithelial cell is induced by high level of endogenous NO in the development of pulmonary fibrosis.