Effect of 25G+pars plana vitrectomy combined with preoperative intravitreal injection of Conbercept on proliferative diabetic retinopathy / 国际眼科杂志(Guoji Yanke Zazhi)

AIM: To investigate the clinical effect of 25G+pars plana vitrectomy(PPV)combined with preoperative intravitreal injection of conbercept in the treatment of patients with proliferative diabetic retinopathy(PDR), and analyze the influence on visual acuity, central foveal thickness(CMT)and serum vascular endothelial growth factor(VEGF)level.METHODS: A retrospective study was conducted from October 2019 to January 2022. A total of 80 patients(87 eyes)with PDR were divided into the two groups according to the treatment method, with 40 patients(45 eyes)treated with 25G+PPV in the control group, and 40 patients(42 eyes)treated with 25G+PPV combined with preoperative intravitreal injection of conbercept in the observation group. The two groups were compared in terms of the best corrected visual acuity(BCVA), intraocular pressure, CMT and serum VEGF level before treatment and at 2wk, 1 and 3mo after treatment. The patients were followed up for 3mo, with postoperative complications and recurrence recorded.RESULTS: The incidence of intraoperative bleeding in the observation group was significantly lower than that in the control group(P<0.05). After treatment, the BCVA of the two groups was improved(P<0.05), CMT and serum VEGF level were decreased(P<0.05), but there was no significant change in intraocular pressure(P>0.05). The BCVA and CMT of observation group were lower than those of control group at 1 and 3mo after treatment(P<0.05). Serum VEGF level in the observation group was lower than that in the control group at 3mo after treatment(P<0.05). The incidence of complications in observation group(5%)within 3mo after treatment was significantly lower than that in control group(18%; P<0.05). There was no statistically significant difference in recurrence rate of PDR between the two groups(P>0.05).CONCLUSION: With few complications, 25G+PPV combined with preoperative intravitreal injection of conbercept is effective in the treatment of patients with PDR, which can better promote postoperative vision recovery, improve macular edema, and reduce serum VEGF level.

Role of inflammatory regulators in neovascularization of diabetic retinopathy / 国际眼科杂志(Guoji Yanke Zazhi)

@#Diabetic retinopathy(DR)is one of the common diseases that can lead to blindness, and its complicated pathogenesis has not been elucidated completely at present. Many studies have emerged that chronic inflammation plays a prominent role in the pathogenesis of DR. That many “hot spots”of inflammatory molecules, such as IL-6, IL-1β, TNF-α, and MCP-1,are involved in the part of essential mechanism of disease through complex and intertwined inflammatory pathways in the recently research. Above inflammatory factors and angiogenic factors will promote each other, especially when pathological neovascularization occur, which is greatly increasing in severity of the disease. The article made a general debate about the effect of inflammation in DR, and the relationship between inflammation and neovascularization.

The effects of tectochrysin on prostate cancer cells apoptosis and its mechanism / 中国应用生理学杂志

OBJECTIVE@#To investigate the effects of tectochrysin on prostate cancer cell line 22Rv.1 and reveal its molecular mechanism.@*METHODS@#Tectochrysin at the concentrations of 0～20 μg/ml was applied to 22Rv.1 cells and normal prostate cell RWPE-1. The proliferation activity of the cells was detected by MTS assay. Flow cytometry and hoechst 33342 staining were used to analyze the effects of drugs on cell apoptosis, death, cell cycle and nuclear type changes. LDH release test was used to analyze the cytotoxicity of the drug to 22Rv.1 cells. QPCR and Western blot were used to analyze the effects of the drug on the expressions of genes in 22Rv.1 cells. Finally, the tumor inhibited effect of the drug on the bearing tumor BALB/c mice were confirmed though anti-tumor experiment.@*RESULTS@#Tectochrysin could significantly inhibit the proliferation activity of 22Rv.1 cells and induced their apoptosis, and promoted the expressions of genes dr4, dr5, trail, p53, caspase-3, caspase-8, caspase-9, bid, bax and foxo3, inhibited the expressions of anti-apoptotic genes akt, pi3k and bcl-2.@*CONCLUSION@#Tectochrysin can induce prostate cancer cells apoptosis through affecting TRAIL and PI3K/AKT signaling pathways, and has anti-prostate cancer effect.

Effects of 2-12alkyl-6-methoxycyclohexa-2, 5-diene-1, 4-dione(DMDD)on diffuse large B lymphoma and its mechanism / 中国应用生理学杂志

OBJECTIVE@#To investigate the effects and molecular mechanisms of 2-12alkyl-6-methoxycyclohexa-2,5-diene-1,4-dione(DMDD) on diffuse large B lymphoma (DLBCL).@*METHODS@#In animal experiments, 4-week-aged BALB/C mice were divided into 5 groups, 20 mice in each group. Mice were inguinal injected with DLBCL cell line OCI-LY19 cells 0.1 ml at the concention of 1 × 10 /ml. Two days later, mice were treated with DMDD at the doses of 0, 1, 5, 25 and 125 mg/kg by intragastric administration respectively, once /2 days. Ten mice of each group were killed on the 18th day of administration, and the tumor tissues were weighed. The survival time of the remaining mice were recorded. In cell experiments, OCI-LY19 cells were added to 96-well culture plates, 100 μl 1×10 cells/ml per well, then 100 μl DMDD was added to the well and the final concentrations were 0, 1, 5, 25 and 125 μmol/L respectively. The cells were treated with DMDD for 0, 24, 48 and 72 h, three wells in each group. The cell proliferation activity was detected by MTS assay. According to the results of cell proliferation experiments, OCI-LY19 cells were treated with DMDD at the concentrations of 0 μmol/L, 5 μmol/L and 25 μmol/L for 24 h. The apoptosis rate was analyzed by flow cytometry, the nuclear type was observed by hoechst staining, the mitochondrial membrane potential was observed by JC-1 staining, cytotoxicity of drugs was evaluated by LDH release experiment, gene expression and transcription were analyzed by qPCR and Western blot.@*RESULTS@#Compared with 0 mg/kg drug group, DMDD at the dose of 1～125 mg/kg could inhibit the growth of tumor tissue in mice and prolong their survival time (P＜0.01). Cell experiments showed: in DMDD group, the proliferation activity of OCI-LY19 cells was decreased significantly and the level of apoptosis was increased significantly (P＜0.01), nuclear fragmentation, agglutination, apoptotic bodies occurred and mitochondrial membrane potential was decreased, the LDH release rate was increased significantly (P＜0.01), the expressions of caspase-3 and bax genes and the phosphorylation level of Ikappa B alpha in cells were up-regulated significantly, the protein expression levels of bcl-2, bcl-xL, jak2 and stat3 were inhibited significantly (P＜0.01).@*CONCLUSION@#DMDD can inhibit the expressions of JAK2, STAT3 and p-Ikappa B alpha in JAK2/STAT3 and NF-kappa B signal pathways, down-regulate BCL-2/BAX and activate Caspase-3, finally, activate the endogenous pathway of mitochondrial apoptosis in OCI-LY19 cells and promote the apoptosis of DLBCL cells, inhibit proliferation of OCI-LY19 cells. It has inhibitive effects on DLBCL.

Anti-hepatoma effects of Smac analogue Birinapant and its related molecular mechanism / 中国应用生理学杂志

OBJECTIVE@#To investigate the effects of Birinapant on hepatocellular carcinoma cells and its related molecular mechanisms.@*METHODS@#Human hepatocellular carcinoma cells QGY-7701 were treated with 0, 1, 5, 25 and 125 nmol/L Birinapant for 24, 48 and 72 hours respectively, each experiment 3 wells.The proliferation activity of cells, the apoptosis levels, the cells nuclear type, the mitochondrial membrane potential, the transcription and expression levels of genes and the cytotoxicity of Birinapant were analyzed.At the same time, 4-week-old male BALB/C mice were randomly divided into 5 groups, with 20 mice in each group.The mice were inguinal injected with QGY-7701 cells, and then subcutaneous injected with Birinapant (concentrations ranging from 0, 1, 5, 25, 125 μg/kg) in each group after two days, once every other day.On 18 day since first Birinapant injection, 10 mice were killed in each group to weigh tumor tissue and survival time was recorded from the remaining 10 mice.The effects of Birinapant on the growth of the tumor and the survival time of tumor-bearing mice were observed.@*RESULTS@#Compared with the negative control (NC) group, the proliferation activity of QGY-7701 was inhibited significantly after Birinapant treatment and the apoptosis levels were increased significantly (<0.01).The cell mitochondrial membrane potential was decreased and the karyotype was changed (<0.01).At the same time, the transcription and expression levels of genes cellular inhibitor of apoptosis protein 1(cIAP-1), cellular inhibitor of apoptosis protein 2(cIAP-2), ras, raf, mek and erk were significantly decreased (<0.01), while the expression levels of caspase-3 and caspase-9 genes were up-regulated (<0.01).Compared with the model group (MG), the growth of the tumor was inhibited significantly and the survival time of the tumor-bearing mice was prolonged after Birinapant treatment (<0.01).@*CONCLUSIONS@#Birinapant can inhibit the expression of cIAP-1, cIAP-2 and the proteins of Ras-Raf-MEK-ERK signal pathways, so as to activate the mitochondria mediated endogenous apoptosis pathway.Birinapant shows a certain inhibitory effect on liver cancer.

Effects of human umbilical cord mesenchymal stem cells in the treatment of paraquat-induced lung injury / 中华劳动卫生职业病杂志

<p><b>OBJECTIVE</b>To evaluate the clinical effect and safety of human umbilical cord mesenchymal stem cells (HUCMSCs) in the treatment of lung injury caused by acute paraquat poisoning.</p><p><b>METHODS</b>Thirteen patients with lung injury caused by acute paraquat poisoning, who were admitted to Guangzhou No. 12 People's Hospital from December 2008 to December 2012, were divided into HUCMSC group (n = 5) and control group (n = 8). All patients received conventional treatment, while the HUCMSC group was treated with HUCMSCs as an addition. Sequential Organ Failure Assessment (SOFA) system, which was created by the Infection Section of European Society of Intensive Care Medicine, and Acute Physiology and Chronic Health Evaluation II were used to acquire the SOFA scores of patients. The lung injury was evaluated with lung injury score (LIS). The two groups were compared with respect to maximum SOFA scores at 1, 3, 5, 7, 14, and 15 days after paraquat poisoning.</p><p><b>RESULTS</b>The HUCMSC group showed significantly lower maximum SOFA scores than the control group at 15d after poisoning (1.80 ± 2.05 vs 13.50 ± 7.59, P < 0.05). The LISs of the HUCMSC group after treatment (0.45 ± 0.27) were significantly lower than those of the HUCMSC group before treatment (1.15 ± 0.34) and those of the control group after treatment (2.94 ± 1.20) (P < 0.01). In the HUCMSC group, all patients survived, and they complained no discomfort and showed normal liver, kidney, and lung functions in reexamination; one patient showed incompletely absorbed shadow in the posterior segment of the left lower lobe of the lung during lung CT scan, and no abnormal findings were seen in other patients. In the control group, one patient survived, and others died. No adverse reactions, such as chill and fever, were presented in the HUCMSC group.</p><p><b>CONCLUSION</b>HUCMSCs show promise for clinical application in the treatment of lung injury caused by acute paraquat poisoning.</p>

One-year continuous observation of change in peripheral T cell subsets in workers exposed to low levels of benzene / 中华劳动卫生职业病杂志

<p><b>OBJECTIVE</b>To observe the T cell subsets and blood cells in the peripheral blood of workers exposed to low levels of benzene for one year, and to investigate the relationship between T cell function impairment and benzene-induced hematopoietic injury after benzene exposure.</p><p><b>METHODS</b>Eighty-eight workers (58 males and 30 females, aged 18 ∼ 22 years) who just began to work in the workshop of a paint factory with exposure to benzene in Guangzhou, China were assigned to experimental group, and 88 workers (58 males and 30 females, aged 18 ∼ 25 years) who worked in the workshop without exposure to benzene were selected as controls. The blood samples of the workers were examined once every 4 months to measure the percentages of peripheral T cell subsets and peripheral blood cell counts in the one-year study. The benzene concentrations at operation points were also measured.</p><p><b>RESULTS</b>The peripheral blood cell counts in the benzene-exposed workers had no significant changes in the first and second examinations; the white blood cell (WBC) counts in the experimental group in the third and fourth examinations were significantly lower than that in the control group [(6.4 ± 3.0)×10(9)/L and (6.3 ± 2.7)×10(9)/L vs (7.3 ± 3.0)×10(9)/L, P < 0.05], and the platelet (PLT) count in the experimental group in the fourth examination was also significantly lower than that in the control group[(179 ± 74)×10(9)/L vs (189 ± 70)×10(9)/L, P < 0.05]. Compared with those in the control group (CD4+: 54.29 ± 12.78%, CD8+: 37.25 ± 12.30%), the percentage of CD3+ T cells in the experimental group increased in the third examination; the percentage of CD4+ T cells in the experimental group decreased continuously in the second, third, and fourth examinations (50.77 ± 11.05%, 45.40 ± 9.41%, and 41.27 ± 10.62%), while the percentage of CD8+ T cells in the experimental group kept increasing (46.07 ± 10.18%, 50.36 ± 10.62%, and 56.40 ± 9.41%) (P < 0.05).</p><p><b>CONCLUSION</b>The change in T cell subsets precedes that in the blood system in the workers exposed to low levels of benzene.</p>

Protective effects of human bone marrow mesenchymal stem cells on hematopoietic organs of irradiated mice / 中国实验血液学杂志

The objective of this study was to explore the protective effects of human bone marrow mesenchymal stem cells (MSC) on hematopoietic organs of irradiated mice. Human bone marrow MSC were isolated, ex vivo expanded, and identified by cell biological tests. Female BALB/c mice were irradiated with (60)Co γ-ray at a single dose of 6 Gy, and received different doses of human MSC and MSC lysates or saline via tail veins. The survival of mice was record daily, and the femurs and spleens were harvested on day 9 and 16 for pathologic examination. The histological changes were observed and the cellularity was scored. The results showed that the estimated survival time of MSC- and MSC lysate-treated mice was comparable to that of controls. The hematopoiesis in the bone marrow of mice that received high-dose (5×10(6)) of MSC or MSC lysates was partially restored on day 9 and the capacity of hemopoietic tissue and cellularity scorings were significantly elevated as compared with that of controls (P < 0.05). Proliferative nudes were also obviously observed in the spleens of mice that received high-dose of MSC or MSC lysates on d 9 after irradiation. The histological structures of the spleen and bone marrow of the mice that received high-doses (5×10(6)) of MSC or MSC lysates were restored to normal, the cell proliferation displayed extraordinarily active. Further, the cellularity scores of the bone marrow were not significantly different between the high-dose MSC and MSC lysate-treated mice. It is concluded that the bone marrow MSC can promote the hematopoietic recovery of the irradiated mice, which probably is associated with the bioactive materials inherently existed in bone marrow cells.

Abnormal increase in CD8(low) T lymphocyte in patients with occupational chronic lead poisoning / 中华劳动卫生职业病杂志

<p><b>OBJECTIVE</b>To analyze the changes in CD8(low) T lymphocyte subsets in patients with occupational chronic lead poisoning.</p><p><b>METHODS</b>Flow cytometric analysis was used to count the numbers of CD8+ cells. 23 patients with occupational chronic lead poisoning and 20 controls were examined.</p><p><b>RESULTS</b>Compared with control group (8.21% ± 3.02%), the CD8(low) T lymphocyte (12.98% ± 5.62%) were significantly increased in patients with occupational chronic lead poisoning.</p><p><b>CONCLUSION</b>Although the ratio of CD+ T lymphocyte is normal, the CD8 level is significantly decreased. The increase of CD8(low) T lymphocyte may be an important phenomenon of immuno-injury induced by lead. CD8(low) T lymphocyte could be an new direction for research of lead immuno-toxicity.</p>

The comparative study of quartz dust and bleomycin-induced pulmonary fibrosis in rats / 中华劳动卫生职业病杂志

<p><b>OBJECTIVE</b>To compare the pulmonary alveolitis and the early fibrosis of pulmonary fibrosis induced by quartz dust and bleomycin in rats, and investigate their mechanism.</p><p><b>METHODS</b>The female rats were divided into three groups: control group exposed to normal saline by the trachea; SiO2 group exposed to SiO2 by the trachea; BLM group exposed to BLM A5 by the trachea. Each half of the animals were sacrificed on the 7th and 14th day after exposure. The lungs of rats were collected to observe pulmonary alveolitis by HE staining and to observe fibrosis by saturated picric acid sirius red staining. The expression of tumor necrosis factor-alpha (TNF-alpha) and CD68 in pulmonary tissues were analyzed quantitatively by immunohistochemistry and image analysis system.</p><p><b>RESULTS</b>(1) The alveolitis and pulmonary fibrosis of rats in both SiO2 group and BLM group were became more serious gradually over time, HE staining under light microscope showed that BLM group on the 7th day had the most obvious alveolitis (2.814 +/- 0.832), the saturated picric acid sirius red staining under polarized light showed that BLM group on the 14th day had the worst pulmonary fibrosis (1284.57 +/- 554.72), which were significantly higher than those (103.69 +/- 18.29 and 111.78 +/- 37.45) in control group and SiO2 group on the 7th day (P < 0.05). (2) The results of immunohistochemistry examination indicated that the expression (17.100 +/- 1.831) of TNF-alpha in the BLM group on the 7th day was significantly higher than those (0.451 +/- 0.441, 7.909 +/- 1.275 and 13.506 +/- 1.454) in control group, SiO2 group on 7th day and BLM group on 14th day (P < 0.05). The expression (22.778 +/- 2.512) of TNF-alpha in the SiO2 group on the 14th day was significantly higher than those in control group, SiO2 group on 7th day and BLM group on 14th day (P < 0.05). The expression (134.941 +/- 35.951) of CD68 in the SiO2 group on the 14th day was significantly higher than those in control group, SiO2 group on 7th day and BLM group on 14th day (P < 0.05).</p><p><b>CONCLUSION</b>The early alveolitis of BLM-induced lung injury model was more serious than that of SiO2-induced lung injury model, and the fibrosis process of BLM-induced lung injury model was earlier than that of SiO2-induced lung injury model. TNF-alpha plays an important role in the course of both models, but macrophages is involved in SiO2-induced pulmonary in a more continuous way than in BLM-induced pulmonary fibrosis.</p>

The experimental study of suppressing silicosis fibrosis / 中华劳动卫生职业病杂志

<p><b>OBJECTIVE</b>To compare the difference of effects on SiO(2)-induced alveolitis and early fibrosis between bone marrow-derived mesenchymal-like stem cells (BM-MSCs) and BM-MSCs transfected by pcDNA3.1-HGF and to explore the mechanism of this effects.</p><p><b>METHODS</b>The Primary BM-MSCs from Wistar male young rats were cultured and labeled by 4, 6-diamidino-2-phenylindole (DAPI). Fifty Wistar rats were randomly divided into 3 groups:model group (10 rats),which was administered with SiO(2) by the trache, the next day,injected PBS via the tail vein; BM-MSCs group (20 rats),which was administered with SiO(2) by the trache, the next day,injected with 1 ml suspension of BM-MSCs via the tail vein; pcDNA3.1-HGF plus BM-MSC group (20 rats),which was administered with SiO(2) by the trache, the next day,injected with 1 ml suspension of BM-MSCs transfected by pcDNA3.1-HGF via the tail vein. On the 14th and 28th days after treatment, half of the animals were sacrificed, respectively, and the lungs were harvested for frozen section to observe the cell marked by DAPI. HE staining under a fluorescent microscope, and to observe the pulmonary alveolitis and fibrosis by HE and Masson staining under a light microscope. Western blot assay was used to detect the expression of HGF in rat lungs. The expression levels of tumor necrosis factor-α (TNF-α) in pulmonary tissues were analyzed quantitatively by ELISA. The contents of HYP in pulmonary tissues were analyzed quantitatively by sample hydrolysis method.</p><p><b>RESULTS</b>On the 14th and 28th days after treatment, the scores of pulmonary alveolitis and early fibrosis in pcDNA3.1-HGF plus BM-MSCs group were 2.36 ± 0.17, 2.8 ± 0.14 and 0.1 ± 0.11, 1.16 ± 0.13, which were significantly lower than those (1.68 ± 0.17, 1.58 ± 0.31 and 0.54 ± 0.15, 1.36 ± 0.13) in BM-MSCs group, also which were significantly lower those (2.36 ± 0.17, 2.80 ± 0.14 and 0.64 ± 0.09, 1.84 ± 0.17) in model group (P < 0.05); On the 14th and 28th days after treatment, the TNF-α contents of pulmonary tissues in pcDNA3.1-HGF plus BM-MSCs group were 280.4 ± 23.11 and 249.78 ± 22.33 pg/mg, which were significantly lower than those (341.58 ± 35.34, 442.29 ± 36.76 pg/mg and 319.51 ± 17.84, 348.53 ± 33.95 pg/mg) in BM-MSCs and model groups (P < 0.05); On the 14th and 28th days after treatment, the HYP contents of pulmonary tissues in pcDNA3.1-HGF plus BM-MSCs group were 0.46 ± 0.04 and 0.65 ± 0.05 µg/mg, which were significantly lower than those (0.63 ± 0.04, 1.04 ± 0.07 µg/mg and 0.72 ± 0.60, 1.39 ± 0.60 µg/mg) in BM-MSCs and model groups (P < 0.05).</p><p><b>CONCLUSION</b>The effects of BM-MSCs transfected by pcDNA3.1-HGF on suppressing pulmonary alveolitis and early fibrosis induced by SiO2 were better than those of BM-MSCs. The mechanism may be associated with the reduced pulmonary inflammation.</p>

Investigating the treatment of silicosis with autologous bone marrow-derived mesenchymal stem cells / 中华劳动卫生职业病杂志

<p><b>OBJECTIVE</b>To explore the safety and curative effects of autologous bone marrow-derived mesenchymal stem cells (BMSCs) in the treatment of silicosis.</p><p><b>METHODS</b>The protocol was approved by the Ethics Committee of the hospital, and ten patients with silicosis who had given written consent were enrolled in this study. BMSCs isolated from 100 ml of bone marrow for each case were purified and cultured. In each case the 3rd generation of qualified BMSCs (5 × 10(7)) were intravenously administered weekly for 3 weeks. Three cases among 10 patients were treated with BMSCs modified by hepatocyte growth factor (HGF) gene. The clinical symptoms, chest films, chest CT, pulmonary functions, T cells, serum IgG and ceruloplasmin (CP) were observed in 6 or 9 months after treatment.</p><p><b>RESULTS</b>No obvious sub-effect was observed in cases treated with BMSCs, the clinical symptoms (such as cough, sputum and chest tightness) basically disappeared in 9 months after treatment. Pulmonary function tests showed that FVC increased from 71.2% ± 17.0% to 84.0% ± 10.9% (P < 0.01) and FEV1.0 increased from 67.5% ± 17.7% to 80.6% ± 14.9% (P < 0.01). The levels of serum CP and IgG significantly decreased (P < 0.01). Further, the chest films and CT in cases treated with autologous BMSCs modified by HGF gene were improved to different extent.</p><p><b>CONCLUSION</b>Treatment with autologous BMSCs modified by HGF gene exhibit a beneficial effect on silicosis.</p>

Hepatocyte growth factor combined with autologous bone marrow mesenchymal stem cell transplantation for treatment of silicosis / 中华劳动卫生职业病杂志

<p><b>OBJECTIVE</b>To evaluate the potential role of hepatocyte growth factor (HGF) combined with bone marrow mesenchymal stem cells (BMSC) autograft for the treatment of silicosis.</p><p><b>METHODS</b>Bone marrow (100 ml) was aspirated from a severe silicosis patient. BMSCs isolated, purified and cultured in vitro. When BMSC came to 70% confluence at passage 3, the culture medium was added liposomes (lipo2000) and plasmid-HGF (p-HGF) and cultured for 2 d. HGF-MSCSs (5 × 10(7) cells) were resuspended in 50 ml 0.9% sodium chloride (NS) and infused Intravenous drip at 3 consecutive times (once a week). Clinical follow-up were performed before and after treatment: (1) pulmonary high-kV X-ray, chest CT examination; (2) pulmonary function test; (3) determination of serum ceruloplasmin.</p><p><b>RESULTS</b>The symptoms such as coughing, chest tightness disappeared at 12 months after treatment. Pulmonary function tests showed significant changes after treatment: forced vital capacity (FVC) increased from 64.6% to 81.0%, forced expiratory volume in one second (FEV(1.0)) increased from 68.7% to 90.1%, 1 second rate (FEV(1.0)/FVC%) reduced from 111.6% to 107.1%, the maximum mid-expiratory flow (FEF(25%∼75%) decreased from 100.2% to 94.6%, forced expiratory vital capacity 75% of the moment bit of gas flow (MEF(75%)) increased from 99.2% to 113.5%, forced expiratory vital capacity 50% of the moment bit of gas flow (MEF(50%)) increased from 125.3% to 130.2%, forced expiratory vital capacity 25% of the moment bit of gas flow (MEF(25%)) reduced from 86.9% to 71.7%; serum ceruloplasmin levels decreased from 690 mg/L to 180.6 mg/L; lung high-kV X-ray at 1st review showed that diffuse lung nodules had been absorbed and getting smaller than before treatment; chest CT showed that the distribution and number of small nodules at double lung fields decreased than before treatment.</p><p><b>CONCLUSION</b>HGF combined with BMSC transplantation may have some potential role for the treatment of silicosis patients.</p>

Therapy of aplastic anemia with autologous peripheral mononuclear cells treated by IL-2 and GM-CSF in culture: a long-term follow-up report on 49 patients / 中国实验血液学杂志

This study was purposed to evaluate the long-term outcome and the safety of autologous peripheral blood mononuclear cells (PBMNC) treated by interleukin 2 (IL-2) and granulocyte-macrophage colony stimulating factor (GM-CSF) in the therapy of patients with aplastic anemia (AA). The therapy of 49 patients admitted BG in hospital from April 2001 to December 2007 were analyzed retrospectively. PBMNC were isolated and cultured for 48 hours in presence of IL-2 and GM-CSF. Cells were collected, and 6 × 10(6) - 1 × 10(8) PBMNC were intravenously injected weekly for 4 - 22 months. Hematopoietic recovery was evaluated by examinations of peripheral blood, bone marrow aspirates and bone marrow biopsy. Flow cytometry was used to assess the peripheral T cell subsets before and after treatment. Polymerase chain reaction was performed to observe the clonal diversity of T cell receptor variable β-chain (TCR-Vβ) recombination. The results showed that 37 cases were cured and none of them relapsed during the follow-up, 5 cases were in partial remission, 3 cases got improvement, and 4 cases showed no response. The total efficiency reached up to 91.8%. The ratios of CD4(+)/CD8(+) subsets were abnormal in 39 patients prior to the treatment, and 31 cases restored to the normal range after cell transfusions. Analysis on the clonal diversity of TCR-Vβ recombination in 11 patients showed the transition from monoclonal or biclonal spectratype to polyclonal one. No long-term side effects were documented. It is concluded that the treatment with PBMNC treated by IL-2 and GM-CSF is generally safe and effective. The underlying mechanisms may be in relation to the restoration of cell immunity.

T-cell receptor V alpha gene repertoire and clonal expansion in benzene-exposed workers / 中华劳动卫生职业病杂志

<p><b>OBJECTIVE</b>To observe the distribution of TCR V alpha gene repertoire and clonal expansion in peripheral blood mononuclear cells from 9 donors and 16 workers exposed to benzene.</p><p><b>METHODS</b>Complementarity determining region 3 (CDR3) of TCR V alpha subfamily genes were amplified using RT-PCR. The PCR products were further analyzed by genescan to evaluate clonality of T cells.</p><p><b>RESULTS</b>Almost all of 29 V alpha subfamily could be detected in 9 donors. 1 approximately 11 V alpha subfamilies were identified in all but one of the workers studied. The most frequently expressed V alpha subfamily were V alpha 3, V alpha 12 and V alpha 19 (68.8%), V alpha 14 (56.3%), with a lower expression rate found in V alpha 5, V alpha 15, V alpha 16, V alpha 22, V alpha 23 and V alpha 24 (6.3%). Clonal expansion T cells in one or more V alpha subfamily were found in 12 out of all workers studied, including oligoclonal, oligoclonal trend and biclonal patterns. The frequency of clonal expansion T cells in V alpha 12, V alpha 14 and V alpha 19 subfamilies were higher than others.</p><p><b>CONCLUSION</b>Skewed distribution and clonal expansion of TCR V alpha subfamily T cells could be found in workers exposed to benzene. V alpha 12, V alpha 14 and V alpha 19 subfamilies may be highly sensitive to benzene exposed. This is the first report of clonal expansion TCR V alpha T cells in the benzene-exposed group. The bias pattern of TCR V alpha T cells may be due to the immune cytotoxicity from benzene. However, whether the oligoclonality in some V alpha subfamilies reflect the phenomenon of clone absence or may be a response clone to benzene-related impairment during exposed to benzene, remains an open question.</p>

Clinical experiment of cytokines induced killer cells for treatment of benzene poisoning / 中华劳动卫生职业病杂志

<p><b>OBJECTIVE</b>To assess the reaction of cytokines induced killer (CIK) cells treatment in hematopoietic injury at different levels on patients with benzene poisoning and seek a novel, safe and effective immunotherapy for benzene poisoning.</p><p><b>METHODS</b>CIK cells were in vitro activated by interleukin-2 (IL-2) and granulocyte-macrophage-colony-stimulating factor (GM-CSF) from the peripheral blood mononuclear cells (PBMC). Thirty-two patients with benzene poisoning were treated with CIK cells. Nineteen patients with mild or moderate benzene poisoning in the control group were treated with VitB4, batilol, leucogen, inosine and stanozolol. The results for treatment of 12 patients with aplastic anemia induced by severe benzene poisoning (the efficacy rate and the case fatality rate) were analyzed. The change of T-lymphocyte subset analyzed by flow cytometry was also observed before and after treatment.</p><p><b>RESULTS</b>For mild or moderate benzene poisoning, the increase of WBC and RLT in CIK group was higher than that in the control group (P < 0.05). The CD(4)/CD(8) levels were significantly increased after CIK treatment. And for severe benzene poisoning, the effective rate of the CIK group was 91.7% and the mortality rate was 0%.</p><p><b>CONCLUSION</b>CIK treatment is safe and effective for hematopoietic injury caused by benzene poisoning. The mechanism may be related with the immune modulation of CIK treatment on immunodeficiency of patients with benzene poisoning.</p>

Effects of lead exposure on thymic output naive T cells function / 中华劳动卫生职业病杂志

<p><b>OBJECTIVE</b>To investigate the levels of T cell receptor rearrangement excision DNA circles (TRECs) within peripheral blood from workers exposed to lead, and thereby to evaluate the number of naive T cells and recent thymic output function.</p><p><b>METHODS</b>Quantitative detection of TRECs in peripheral blood mononuclear cells (PBMNC) from 10 cases of workers exposed to lead was performed by real time PCR analysis. 11 workers without exposure to lead served as unexposed controls. In addition, the relationship between TRECs, age, length of service, blood lead, urea lead, blood ZPP and urea delta-ALA was investigated.</p><p><b>RESULTS</b>The mean value of TRECs in workers exposed to lead was (2.44 +/- 1.87)/1000 PBMC, significantly under (5.60 +/- 3.96)/1000 PBMC in unexposed controls. A significant negative correlation was found between the TRECs and urea-ALA. But there was no significant correlation between them after controlling for blood lead, urea lead.</p><p><b>CONCLUSION</b>Lead exposure may damage thymic output naive T cells function. Furthermore, low-level exposure to lead may damage immune system and earlier than expected.</p>

Changes of T lymphocyte subsets in workers with long-term benzene exposure / 中华劳动卫生职业病杂志

<p><b>OBJECTIVE</b>To observe the changes of T-lymphocyte subsets in workers with long-term benzene exposure, and further understand the benzene's lymphotoxicity.</p><p><b>METHODS</b>Blood was sampled from 44 patients with chronic benzene poisoning of different degrees, (mild 22 patients, moderate 14, severe 8) respectively. Twenty-two health benzene exposed workers, and 94 health unexposed workers served as normal control. A total of the phenotype (CD4, CD8) of T lymphocyte in peripheral blood was analyzed by indirect immunofluorescence assay.</p><p><b>RESULTS</b>Lymphocyte subset analysis showed significantly decreased CD4(+) T lymphocytes, CD4(+)/CD8(+) ratio, except CD8(+) T lymphocytes in benzene exposed groups (P<0.05). Among the four benzene-exposed groups, CD4(+) T lymphocytes and CD4(+)/CD8(+) ratio showed no difference (P>0.05).</p><p><b>CONCLUSION</b>The primary changes of T-lymphocyte subsets in workers following benzene long-term exposure are the decrease of CD4(+)%, but the changes are not correlated with haematopoietic injury.</p>

Changes in T lymphocyte subsets and plasma Th1/Th2 cytokine levels in patients with occupational chronic lead poisoning / 中华劳动卫生职业病杂志

<p><b>OBJECTIVE</b>To investigate the changes in T lymphocyte subsets and plasma Th1/Th2 cytokine levels in patients with occupational chronic lead poisoning.</p><p><b>METHODS</b>Flow cytometric analysis was used to count the numbers of CD3+, CD4+, CD8+ cells and cytometric bead array CBA capture technique was used to quantify plasma IL-2, IL-4, IL-6, IL-10, TNF-alpha, IFN-gamma concentrations. Twenty-three patients with occupational chronic lead poisoning and 20 controls were examined.</p><p><b>RESULTS</b>Compared with control group, CD3+ CD4+ lymphocytes and CD4/CD8 ratios decreased in patients with occupational chronic lead poisoning. The plasma IL-2 level were significantly raised while the IL-10 and IFN-gamma were reduced in patients with occupational chronic lead poisoning.</p><p><b>CONCLUSIONS</b>T lymphocyte subsets and plasma Th1/Th2 cytokine levels could be a new parameter to assess lead immunotoxicity.</p>

Significant decrease of recent thymic output function in patients with severe benzene intoxication / 中国实验血液学杂志

The aim of the present study was to investigate the level of T-cell receptor excision DNA circles (TREC) in peripheral blood mononuclear cells (PBMNC) of patients with severe benzene poison, thereby to evaluate the content of naive T cells and the recent thymic output function. Quantitative detection of TREC in DNA of PBMNCs from 16 normal individuals and 8 cases with severe benzene poison was preformed by real-time PCR using TaqMan technique. The results showed that TREC level was 6.69 +/- 4.79/1 000 PBMNCs in normal individuals, however, significant decrease was shown in patients with severe benzene poison (1.03 +/- 0.44/1 000 PBMNCs, P < 0.01). The TREC level was persistently low in period of benzene poisoning, even if peripheral blood cell counts were at normal levels. In conclusion, the recent thymic output function is remarkably decreased in patients with severe benzene intoxication, that may obviously damage the T cell immune function.