Current status of diagnosis and treatment of chronic lymphocytic leukemia in China: A national multicenter survey research / 中华血液学杂志

Objective: To understand the current status of diagnosis and treatment of chronic lymphocytic leukemia (CLL) /small lymphocytic lymphoma (SLL) among hematologists, oncologists, and lymphoma physicians from hospitals of different levels in China. Methods: This multicenter questionnaire survey was conducted from March 2021 to July 2021 and included 1,000 eligible physicians. A combination of face-to-face interviews and online questionnaire surveys was used. A standardized questionnaire regarding the composition of patients treated for CLL/SLL, disease diagnosis and prognosis evaluation, concomitant diseases, organ function evaluation, treatment selection, and Bruton tyrosine kinase (BTK) inhibitor was used. Results: ①The interviewed physicians stated that the proportion of male patients treated for CLL/SLL is higher than that of females, and the age is mainly concentrated in 61-70 years old. ②Most of the interviewed physicians conducted tests, such as bone marrow biopsies and immunohistochemistry, for patient diagnosis, in addition to the blood test. ③Only 13.7% of the interviewed physicians fully grasped the initial treatment indications recommended by the existing guidelines. ④In terms of cognition of high-risk prognostic factors, physicians' knowledge of unmutated immunoglobulin heavy-chain variable and 11q- is far inferior to that of TP53 mutation and complex karyotype, which are two high-risk prognostic factors, and only 17.1% of the interviewed physicians fully mastered CLL International Prognostic Index scoring system. ⑤Among the first-line treatment strategy, BTK inhibitors are used for different types of patients, and physicians have formed a certain understanding that BTK inhibitors should be preferentially used in patients with high-risk factors and elderly patients, but the actual use of BTK inhibitors in different types of patients is not high (31.6%-46.0%). ⑥BTK inhibitors at a reduced dose in actual clinical treatment were used by 69.0% of the physicians, and 66.8% of the physicians had interrupted the BTK inhibitor for >12 days in actual clinical treatment. The use of BTK inhibitors is reduced or interrupted mainly because of adverse reactions, such as atrial fibrillation, severe bone marrow suppression, hemorrhage, and pulmonary infection, as well as patients' payment capacity and effective disease progression control. ⑦Some differences were found in the perceptions and behaviors of hematologists and oncologists regarding the prognostic assessment of CLL/SLL, the choice of treatment options, the clinical use of BTK inhibitors, etc. Conclusion: At present, a gap remains between the diagnosis and treatment of CLL/SLL among Chinese physicians compared with the recommendations in the guidelines regarding the diagnostic criteria, treatment indications, prognosis assessment, accompanying disease assessment, treatment strategy selection, and rational BTK inhibitor use, especially the proportion of dose reduction or BTK inhibitor discontinuation due to high adverse events.

Regulation of TRIP13 on Proliferation and Apoptosis of B-Cell Lymphoma Cells and Its Mechanism / 中国实验血液学杂志

OBJECTIVE@#To explore the regulatory effect of TRIP13 on the proliferation and apoptosis of B-cell lymphoma cells and its possible molecular mechanism by knocking down/overexpressing TRIP13 on the cell lines Granta-519 and JVM-2.@*METHODS@#Lentiviral transfection technology was used to construct Granta-519 and JVM-2 cells with knocked down or overexpressed TRIP13 and their control cells. The efficiency of transfection was determined by fluorescence microscopy. The efficiency of knockdown and overexpression was evaluated by real-time quantitative PCR and Western blot. The proliferation was detected by CCK-8 assay. The apoptosis was detected by the Annexin V-APC single staining. The cell cycle was detected by the PI staining. The expression levels of P53, MDM4, and BCL-2 were evaluated by Western blot.@*RESULTS@#After TRIP13 was knocked down, the proliferation ability of Granta-519 and JVM-2 cells was significantly reduced, and the apoptosis rate significantly increased. After TRIP13 was overexpressed, the proliferation ability of Granta-519 and JVM-2 cells was significantly enhanced, and the apoptosis was significantly reduced. After TRIP13 was knocked down, Granta-519 cells had obvious G@*CONCLUSION@#TRIP13 promotes the proliferation of B-cell lymphoma cells, inhibits their apoptosis, and affects their proliferation and apoptosis by participating in the regulation of the cell cycle. TRIP13 promotes the expression of BCL-2 proteins and inhibits the expression of MDM4 protein in B-cell lymphoma cells.

Determination of formononetin and harmful elements of heavy metals in Jinji Pill / 国际中医中药杂志

Objective:To use the high performance liquid chromatography method to determine the content of formononetin in Jinji Pills and by using atomic absorption spectrophotometry,method to determine the harmful elements of heavy metal in Jinji Pills in orer to provide the scientific foundation for improving its quality standards and safety evaluation. Methods:Use Waters XBridge? C18 column (4.6 mm × 250 mm, 5 μm), set mobile phase at acetonitrile-1% phosphoric acid solution (27:73), flow rate 1.0 ml/min, column temperature 30 ℃, detection wavelength 249 nm, column temperature 30 ℃; Lead (Pb) and cadmium (Cd) was detected by graphite furnace method; arsenic (As) was detected by cold steam series graphite furnace method; copper (Cu) was detected by flame method; mercury (Hg) was detected by cold steam method.Results:The formononetin had a good linear relationship between 0.02-2.01 μg, the recovery rate was 98.5%, RSD was 1.53%. Lead (Pb) recovery rate was 103.6%, cadmium (Cd) recovery rate was 95.7%, arsenic (As) recovery rate was 92.4%, mercury (Hg) recovery rate was 104.9%, copper (Cu) recovery rate was 112.5%. Conclusion:This method is of accuracy, specificity, high sensitivity and good reproducibility, which could provide strong evidence for quality improvement and safety use of Jinji pill.

Usefulness of large-section cytokeratin 20 in the detection of intestinal wall infiltration and mesangial metastasis in patients with middle and lower rectal cancer

Objective: The objective of the study was to evaluate the usefulness of large-section cytokeratin 20 (CK20) staining technique in the detection of infiltration on the distal wall and mesangial metastasis in patients with middle and lower rectal cancer. Materials and Methods: A total of 62 patients with rectal cancer in the middle and lower segment were studied on large slices stained with CK20. Logistic regression was used to analyze the clinicopathologic factors related to distal low and middle rectal cancer metastasis to the mesorectum and rectal wall. Results: Two types of distal metastasis of the tumor were observed in the rectal wall in 18% (11/62) of the patients: submucosal invasion and muscularis propria invasion. The extent of distal metastasis to the rectal wall was around 0.5–1.0 cm. Four types of distal metastasis occurred in the mesorectum: lymph node invasion, blood and lymphatic vessel invasion, perineural invasion, and isolated neoplastic microfoci. Distal metastasis to the mesorectum was observed in 24% (15/62) of the patients. The extent of metastasis to the mesorectum was around 0.5–4.0 cm. Another three patients with microcapillary invasion in the distal mesorectum were observed by immunohistochemistry, as it was difficult to determine the spread by conventional hematoxylin and eosin staining. Conclusion: The large-section CK20 staining technique is useful for the detection of infiltration on the distal wall and mesangial metastasis in patients with middle and lower rectal cancer

Drug therapy for non-alcoholic fatty liver disease / 中国实用内科杂志

Non-alcoholic fatty liver disease(NAFLD) is the most common chronic liver disease in the world, but currently there are no approved effective drugs for NAFLD. At present, healthy diet and lifestyle intervention are the basic treatment for NAFLD, but drug therapy, mainly for metabolic syndrome and liver injury, is also essential. Currently, the commonly used drugs include hepatic protectant,insulin sensitizer, lipid-modulating drug, antioxidant, slimming drug, and intestinal probiotics, etc. A variety of new drugs targeting different pathogenesis of NAFLD have been put into clinical trials. In this paper, the current status of drug treatment and research on NAFLD in recent years was summarized.

Prognostic significance of miRNA-223 targeting SOX11 in mantle cell lymphoma / 中华血液学杂志

Objective: To explore the expression and prognostic significance of miR-223 in patients with mantle cell lymphoma (MCL) and to investigate the possible mechanism. Methods: Twenty-one newly diagnosed MCL patients with bone marrow involvement were enrolled in the present study, 20 healthy donors as normal control. The expression level of miR-223 and SOX11 mRNA was determined by RQ-PCR. CCK-8 and flow cytometer assays were used to analyze cell proliferation, cell cycle and apoptosis of the constructed miR-223 overexpressing MCL cell line, Granta519 cells. SOX11 protein expression level was determined by Western blot. The target gene of miR-223 was confirmed by dual luciferase reporter assay. Results: ①Of the 21 newly diagnosed MCL patients, 15 were male and 6 female, the median age was 58 (37-72) years. The expression level of miR-223 was significantly down regulated in MCL patients compared with that of healthy donors (14.7±10.5 vs 1 244.1±1 935.2, P<0.001). The lower expression of miR-223 was inversely correlated with high-risk mantle international prognostic index (P=0.001), elevated LDH (P=0.001), ECOG score ≥2 (P=0.035). ②Using the median relative expression level of miR-223 as the cutoff value, 21 MCL patients were divided into high-expression group (n=10) and low-expression group (n=11) and found that the high-expression group had a significantly superior OS (median OS: 36 vs 12 months, P=0.021). ③In vitro results showed that compared with the control group, the proliferation of miR-223 overexpressed Granta519 cells was inhibited (the most significant reduction on 96h, P<0.001), manifested by lower proportion of cells in G2/M phase (P<0.001) and increased apoptosis (P<0.001), and the expression level of SOX11 protein in Granta519 cells was significantly lower than that of the control group. ④miR-223 could inhibited the 3' untranslated region of SOX11, and the expression level of miR-223 was significantly negatively correlated with mRNA level of SOX11 in MCL patients (r=-0.81, P<0.001). Conclusions: The expression of miR-223 was repressed in MCL and was associated with poor clinical outcomes, which may be probably attributed to its direct targeting SOX11.

Changes in Hydrogen Sulfide in Rats with Hepatic Cirrhosis in Different Stages / 华中科技大学学报（医学）（英德文版）

This study aimed to observe changes in the hydrogen sulfide (H2S) system in the blood and liver tissue of rats with hepatic cirrhosis at different stages by studying the effect of H2S on the course of hyperdynamic circulation in rats with hepatic cirrhosis.H2S concentration in the blood from the portal vein and inferior vena cava of hepatic cirrhosis rat model induced with carbon tetrachloride was detected on the 15th,30th,and 52nd day.The expression of cystathionine β-synthase (CBS) and cystathionine γ-lyase (CSE) protein,and CBS and CSE mRNA in the liver was detected by immunohistochemistry and reverse transcriptase polymerase chain reaction (RT-PCR),respectively.The results indicated that H2S concentration in the blood from the portal vein and inferior vena cava of rats with hepatic cirrhosis was significantly lower than that in the control group.H2S was gradually decreased with the development of the disease and significantly lower in the blood from portal vein than in the blood of inferior vena cava at the mid-stage and the late stage groups.The expression levels of CBS and CSE protein,and CBS and CSE mRNA in the livers with hepatic cirrhosis at different stages were all higher than those in the control group,and the expression gradually increased with the development of the disease.The expression of CBS was lower than CSE in the same stages.The results indicated that the CSE mRNA was expressed predominantly in the cirrhosis groups as compared with CBS rnRNA.Among experimental rats,the H2S system has an important effect on the occurrence and development of hyperdynamic circulation in rats with hepatic cirrhosis.This finding adds to the literature by demonstrating that H2S protects vascular remodelling in the liver,and that CSE is indispensable in this process.

Application of lenalidomide in chronic lymphocytic leukemia / 中国实验血液学杂志

In recent years, the incidence of chronic lymphocytic leukemia (CLL) is increasing. Microenvironment and immune system play a key role in the pathogenesis of CLL. The immune system is aggravated by the use of chemotherapeutic agents, such as fludarabine and cyclophosphamide with rituximab(FCR) which are the current standards in frontline therapy. This leads to an increase of infection incidence in patients, resulting in a poor prognosis. The present situation was changed by lenalidomide. Recent studies indicated that lenalidomide monotherapy in treatment of refractory or relapsed CLL patients, the overall response rate(ORR) reached about 32%-47%, CR roughly was 7%-13%; when lenalidomide and rituximab were combined for treatment of refractory or relapsed CLL patients, the ORR reached about 53%-66%, CR about 12%-13%. Moreover, when lenalidomide and ofatumumab were combined, the efficacy is improved significantly and the adverse reactions are greatly reduced. The adverse reactions are neutrophilic granulocytopenia, thrombocytopenia, anemia, tumor lysis syndrome(TLS), tumor flare reaction(TFR) and venous thromboembolism(VTE). This review focuses on the related studies and the latest progress about lenalidomide in CLL.

Efficacy and safety of tauroursodeoxycholic acid in the treatment of liver cirrhosis: a double-blind randomized controlled trial / 华中科技大学学报（医学）（英德文版）

No direct comparison of tauroursodeoxycholic acid (TUDCA) and ursodeoxycholic acid (UDCA) has yet been carried out in the treatment of liver cirrhosis in China. We designed a double-blind randomized trial to evaluate the potential therapeutic efficacy of TUDCA in liver cirrhosis, using UDCA as parallel control. The enrolled 23 patients with liver cirrhosis were randomly divided into TUDCA group (n=12) and UDCA group (n=11), and given TUDCA and UDCA respectively at the daily dose of 750 mg, in a randomly assigned sequence for a 6-month period. Clinical, biochemical and histological features, and liver ultrasonographic findings were evaluated before and after the study. According to the inclusion criteria, 18 patients were included in the final analysis, including 9 cases in both two groups. Serum ALT, AST and ALP levels in TUDCA group and AST levels in UDCA group were significantly reduced as compared with baseline (P<0.05). Serum albumin levels were significantly increased in both TUDCA and UDCA groups (P<0.05). Serum markers for liver fibrosis were slightly decreased with the difference being not significant in either group. Only one patient in TUDCA group had significantly histological relief. Both treatments were well tolerated and no patient complained of side effects. It is suggested that TUDCA therapy is safe and appears to be more effective than UDCA in the treatment of liver cirrhosis, particularly in the improvement of the biochemical expression. However, both drugs exert no effect on the serum markers for liver fibrosis during 6-month treatment.

Efficacy and safety of tauroursodeoxycholic acid in the treatment of liver cirrhosis: a double-blind randomized controlled trial / 华中科技大学学报（医学）（英德文版）

No direct comparison of tauroursodeoxycholic acid (TUDCA) and ursodeoxycholic acid (UDCA) has yet been carried out in the treatment of liver cirrhosis in China. We designed a double-blind randomized trial to evaluate the potential therapeutic efficacy of TUDCA in liver cirrhosis, using UDCA as parallel control. The enrolled 23 patients with liver cirrhosis were randomly divided into TUDCA group (n=12) and UDCA group (n=11), and given TUDCA and UDCA respectively at the daily dose of 750 mg, in a randomly assigned sequence for a 6-month period. Clinical, biochemical and histological features, and liver ultrasonographic findings were evaluated before and after the study. According to the inclusion criteria, 18 patients were included in the final analysis, including 9 cases in both two groups. Serum ALT, AST and ALP levels in TUDCA group and AST levels in UDCA group were significantly reduced as compared with baseline (P<0.05). Serum albumin levels were significantly increased in both TUDCA and UDCA groups (P<0.05). Serum markers for liver fibrosis were slightly decreased with the difference being not significant in either group. Only one patient in TUDCA group had significantly histological relief. Both treatments were well tolerated and no patient complained of side effects. It is suggested that TUDCA therapy is safe and appears to be more effective than UDCA in the treatment of liver cirrhosis, particularly in the improvement of the biochemical expression. However, both drugs exert no effect on the serum markers for liver fibrosis during 6-month treatment.

Effects of cAMP-response element binding protein-1 (CREB-1) on transforming growth factor-b3 (TGFb3) mRNA expression and promoter activity in hepatic stellate cells / 中华肝脏病杂志

<p><b>OBJECTIVE</b>To investigate the effects of cAMP-response element binding protein-1 (CREB-1) on transforming growth factor-b3 (TGF b3) mRNA expression and promoter activity in hepatic stellate cells (HSCs).</p><p><b>METHODS</b>Freshly isolated HSCs from rats were divided into six groups: CREB-1 expression plasmid transfected group (C), siRNA-CREB-1 plasmid transfected group (S), negative control group (N), forskolin treated group (F), H-89 treated group (H), and blank group (B). Rats in each group were further sub-divided according to whether (+) or not (-) they were exposed to exogenous TGF b3. TGF b3 mRNA expression was measured by real time quantitative PCR. HSCs of the C, S, N, F, H and B groups were transfected with the TGF b3 promoter luciferase reporter plasmid (PGL3-TGF b3-P; W group), the TGF b3 promoter luciferase reporter plasmid with CRE mutation (PGL3-basic-TGF b3P-mCRE; M group) and the renilla luciferase control plasmid (pRL-SV40; control group). TGF b3 promoter activity was assessed by luciferase reporter assays.</p><p><b>RESULTS</b>Compared to N(-), the TGF b3 mRNA expression was reduced to 0.69+/-0.15 in S(-) (P less than 0.05) and increased to 4.68+/-2.76 in C(-) (P more than 0.05). Compared to B(-), the TGF b3 mRNA expression was reduced to 0.57+/-0.08 in H(-) (P less than 0.05). The differences between N(+) and N(-), S(+) and S(-), B(+) and B(-), and H(+) and H(-) were all significant (P less than 0.05). The values of TGF b3 promoter activity in S(W), N(W), and C(W) were 0.062+/-0.013, 0.122+/-0.011, and 0.165+/-0.016 (P less than 0.05), but the changes of TGF b3 promoter activity in S(M), N(M), and C(M) were not significant (P more than 0.05). The values of TGF b3 promoter activity in H(W), B(W), and F(W) were 0.154+/-0.010, 0.188+/-0.016, and 0.276+/-0.031 (P less than 0.05), but the changes of TGF b3 promoter activity in H(M), B(M), and F(M) were not significant (P more than 0.05).</p><p><b>CONCLUSION</b>Increased levels of CREB-1 mRNA or p-CREB-1 up-regulate the TGF b3 mRNA expression and promoter activity in rat HSCs. The CRE site in the TGF b3 promoter is critical for this effect, and the gene's activity becomes significantly decreased when the site is missing. Exogenous TGF b3 enhances expression of endogenous TGF b3 in rat HSCs.</p>

Effects of exogenous TGF-β3 on the expression of endogenous TGF-β3 in hepatic stellate cell-T6 (HSC-T6) / 中华肝脏病杂志

<p><b>OBJECTIVE</b>To investigate the effects of exogenous TGF-β3 on the expression of endogenous TGF-b3 in hepatic stellate cell (HSC).</p><p><b>METHODS</b>HSCs were cultured and divided into two groups: TGF-β3 group and blank control group, the cells of TGF-β3 group were exposed to TGF-b3 (10 ng/ml), whereas the blank control group was not treated. The cells were incubated in the presence of exogenous TGF-β3 and then (1) were harvested at 0h, 1h, 2h, 4h, 12h, 24h, and real time PCR was performed to detect the mRNA expression of endogenous TGF-β3. (2) The cells were collected at 0h, 1h, 6h, 12h, and western-blot was used to detect the protein synthesis of endogenous TGF-β3 in HSC; (3) The cell culture supernatant was harvested at 0h, 1h, 2h, 4h, 8h, 14h, 24h, and ELISA was performed to measure the total protein of extracellular TGF-β3; HSCs were treated with TGF-β3 (10 ng/ml) for 2h. The cells were then incubated in serum-free medium and the cell culture supernatant was harvested at 2.25h, 2.5h, 3h, 4h, 6h, 10h and 14h. ELISA was used to detect the extracellular secret ion of endogenous TGF-β3 by HSCs.</p><p><b>RESULTS</b>(1) Exogenous TGF-β3 treatment induced a marked increase in TGF-β3 mRNA expression. By 2h of exogenous TGF-β3 treatment, maximal TGF-β3 mRNA expression levels (2.796 ± 0.518) of 2.74 fold above control values (1.022 ± 0.038) was reached (P < 0.05). Thereafter, TGF-β3 mRNA expression level declined, and the expression level was maintained at level of 1.45-fold for at least 10h and was 1.18-fold above control values by 24h TGF-β3 treatment (P < 0.05); (2) No significant difference about the intracellular protein expression level of endogenous TGF-β3 was found between two groups. (P > 0.05); (3) The total expression level of TGF-β3 reached a peak [(18.931 ± 2.904) ng/ml] at 4h after TGF-β3 treatment (1.89-fold higher than basic TGF-β3 (10 ng/ml). After that, it slowly declined. The expression peak [(0.835 ± 0.027) ng/ml] induction of extracellular secreted TGF-β3 was at 3h (32.12-fold higher than control [(0.026 ± 0.022) ng/ml], (P < 0.05). Thereafter, TGF-β3 slowly decreased after the peak time, and their expressions were still statistically significant as compared to the control (P < 0.05).</p><p><b>CONCLUSION</b>Exogenous TGF-β3 could increase the expression of endogenous TGF-β3 mRNA and extracellular secreted TGF-β3 protein obviously.</p>

Overrepresentation of specific gene segments of expressed immunoglobulin heavy chain variable region among unmutated and mutated patients with chronic lymphocytic leukemia / 中华血液学杂志

<p><b>OBJECTIVE</b>To investigate the overrepresentation of specific gene segments of immunoglobulin heavy chain variable region (IgVH) among unmutated and mutated chronic lymphocytic leukemia (CLL) patients and its prognostic implication.</p><p><b>METHODS</b>Multiplex PCR was used to identify the expression of IgVH segment and its mutation status in CLL.</p><p><b>RESULTS</b>Analyses were successfully performed in 80 of 85 samples. Marked skewed IgVH families were disclosed. The most commonly used VH was VH3 (40.0%), followed by VH4 (30.0%), VHI (13.8%), VH2 (10.0%) and VH5, VH7 (2.5%). Fifty-six patients (70.0%) had mutated VH, 24 (30.0%) unmutated VH. Nine cases (11.3%) were with 100% germline sequence. Fifteen cases (15/24, 62.5%) in VH4, 29 (29/32, 90.7%) in VH3, and 4 (4/11, 36.3%) in VH1 had mutated VH. The most frequently used IgVH gene was VH4-39 (13.8%), and VH4-34 (8.8%). J4 (36/66, 54.5%) and D3 (25/66, 37.8%) were the most frequently used in J and D genes. The progression-free survival (PFS) was 82 and 17 months (P = 0.000), and the overall survival (OS) was 90 and 41 months (P = 0.009), respectively, for mutated and unmutated cases. Recurrent CDR3 sequences were found in our patients and 2 patients with VH1-69 had CDR3 sequences highly similar to those reported in literature.</p><p><b>CONCLUSION</b>There is difference in IgVH gene segment usage and mutational status in different area CLL patients. Recurrent CDR3 sequences were found in specific IgVH gene segments, which highlights the importance of immunoglobulin mediated stimulation in the development of CLL.</p>

The expression of microRNA-155 and microRNA-146a and its' clinical value in chronic lymphoproliferative disorders / 中华血液学杂志

<p><b>OBJECTIVE</b>To investigate the expression of microRNA-155 and microRNA-146a in the CD19(+) B cells of chronic lymphocytic leukemia (CLL), mantle cell lymphoma (MCL), splenic marginal zone lymphoma (SMZL), and to analyze its clinical significance.</p><p><b>METHODS</b>Peripheral blood (PB) (78 cases) and bone marrow (BM) samples (9 cases) from 53 CLL patients, 13 MCL patients, 19 SMZL patients, and 12 healthy donors were collected. Mononuclear cells were isolated and B cells were purified with a CD19(+) magnetic-bead system. Total RNA was extracted from purified CD19(+) cells and microRNAs expression were measured using the TaqMan microRNA quantitative PCR. The results combined with the clinic data of patients were analysed.</p><p><b>RESULTS</b>(1) The expression of microRNA-155 in CLL (4.49 ± 0.83) was significantly higher than in MCL (3.83 ± 0.45) and SMZL (3.80 ± 0.61) (P < 0.05); (2) The level of microRNA-146a in SMZL (3.81 ± 0.59) was significantly higher than in CLL (2.58 ± 0.90) and MCL (2.27 ± 0.88) (P < 0.01); (3) The level of microRNA-155 was significantly higher in IgVH unmutated patients than in mutated patients in CLL (P = 0.012); (4) The microRNAs expression had no statistical difference between two prognostic groups in CLL.</p><p><b>CONCLUSION</b>(1) The expression of microRNA-155 and microRNA-146a is different in malignant lymphoproliferative disorders (LPD); (2) Deregulation of the microRNAs expression might play a critical role in the pathogenesis and prognosis in the LPD.</p>

Application of LMA in patients with acute severe asthma emergency / 中国综合临床

Objective To investigate the clinical value of laryngeal mask airway (LMA) in patients with a-cute severe asthma(ASA). Methods 32 patients with ASA treated with LIMA or mouth-nose mask during 2002 -2009 in our hospital were retrospectively analyzed. Those treated with laryngeal mask airway was taken as observation group and those with Mouth-nose mask as control group. Results The period to oxygen saturation in arterial blood, the time to remove ventilator, and the time to disease improvement in the observation group (389.63±32.82)s, (19.31±2.26) hours,(16.22±3.85) hours were different from that in control group (467.36±41.15) s, (25.18±3.73) hours,(23.66±2.38) hours (P<0.01). After non-invasive positive pressure ventilation, PaCO_2 decreased, PaO_2 and pH increased at 3 and 12 hours in the observation group (P<0.05 or 0.01) from that before treatment. PaCO_2 and pH at 3 hours in the control group were no significant difference before and after treatment (P > 0.05),with an exception of PaO_2 (P < 0.05). PaCO_2, PaO_2 and pH were significantly different (P < 0.05) at 12 hours after treatment from those before treatment. Conclusions LMA should be considered in the selection of non-invasive positive-pressure ventilation (NIPPV) in patients with ASA, for a better improvement of ventilation ef-fectivenoss and accelerating the mitigation of clinical manifestations.

Analysis of efficacy and related factors in 85 chronic myeloid leukemia patients treated with imatinib mesylate / 中国实验血液学杂志

The objective of this study was to evaluate the efficacy of Imatinib on patients with chronic myeloid leukemia (CML) in chronic phase and to analyze its influencing factors. 85 patients received Imatinib mesylate at a dose of 300-600 mg orally per day, and were evaluated for hematologic, cytogenetic, and molecular responses. The results showed that the median follow-up was 21 (range 9 - 78) months. Cumulative complete hematological remission (CHR) rate was 100%, major cytogenetic remission (MCyR) rate was 80%, complete cytogenetic remission (CCyR) rate was 67.1% and complete molecular remission (CMoR) rate was 36.4%. The median time to complete hematological remission (CHR) was 1 (range 1 - 3) month, to complete cytogenetic remission (CCyR) was 6 (range 1 - 24) months. The estimated overall survival rates for patients who received Imatinib for 1, 2, 3 years were (98.7 +/- 1.3)%, (96.5 +/- 2.5)% and (90.1 +/- 6.6)% respectively. The estimated progression-free survival rates at 1, 2, 3 years were (97.6 +/- 1.6)%, (96.1 +/- 2.2)% and (90.0 +/- 1.4)% respectively. The CHR, MCyR and CCyR between low risk, intermediate risk and high risk groups according to the Sokal scoring system and between primarily treated and retreated groups all had no difference. The overall survival of patients who achieved MCyR or CCyR was better than that in patients only achieved hematologic remission (p = 0.026), but there was no significant difference in progression-free survival between them. Univariate analysis for efficacy of Imatinib mesylate revealed that WBC count < 100 x 10(9)/L (p = 0.024), Hb level > or = 130 g/L (p = 0.036), and peripheral basophil count < or = 0.05 (p = 0.024) before therapy were independent favourable factors for achieving MCyR or CCyR. It is concluded that the patients with CML in chronic phase treated with Imatinib can achieve the best hematologic remission and higher cytogenetic remission, it should be considered that the imatinib is a drug of the first-line therapy for untreated and treated patients with CML.

Effects of transforming growth factor beta 3 on the histopathology and expression of collagen I in experimental hepatic fibrotic rats / 中华肝脏病杂志

<p><b>OBJECTIVE</b>To investgate the effects of TGFbeta3 on rat hepatic fibrosis.</p><p><b>METHODS</b>The TGFbeta3 cDNA was cloned into rAAV2 vector. Rats were randomly divided into four groups: normal control group, model group, negative control group and TGFbeta3 group. Hepatic fibrosis was induced by hypodermic injection of 40% CCl(4). Recombinant AAV2-TGFbeta3 viral particles were injected via the vena caudalis one week before CCl(4) treatment. Rats were executed 8 weeks after CCl(4) treatment, global histological change was observed after HE staining, the distribution of the collagen fibers was observed after masson staining, histochemistry was done to observe the expression of collagen I; The positive area rate of the collagen fibers and the average optical rate of collagen I were quantified.</p><p><b>RESULTS</b>HE staining indicated that collagen fibers were reduced in the TGFbeta3 group. Masson staining shown that the collagen fibers were distributed around the blood vessel, in the portal area and disse space. Compared to the model group (13.2%+/-2.2%) and negative control group (12.3%+/-1.5%), the collagen fibers in liver tissues of TGFbeta3 group (7.7%+/-1.5%) were significantly decreased (q = 9.456, P < 0.01; q = 8.217, P < 0.01). Histochemistry indicated that the collagen fibers of liver tissues of TGFbeta3 group (0.185+/-0.033) were significantly higher than those in the model group (0.252+/-0.042) and the negative control group (0.230+/-0.029), (q = 6.228, P < 0.01; q = 4.346, P < 0.01).</p><p><b>CONCLUSION</b>TGFbeta3 alleviates the damage to hepatic cell and the level of fibrosis in CCl(4) treated rats and inhibits the expression of collagen I.</p>

Effects of transforming growth factor-beta 3 gene transfer on type I collagen synthesis of hepatic stellate cells / 中华肝脏病杂志

<p><b>OBJECTIVE</b>To observe the effects of transforming growth factor-beta 3 gene transfer on type I collagen synthesis of cells of HSC-T6.</p><p><b>METHODS</b>Transforming growth factor-beta 1 expression plasmid and transforming growth factor-beta 3 expression plasmid were constructed. The recombinant expression plasmids pcDNA3.1(+)-TGF beta 1 and pcDNA3.1(+)-TGF beta 3 were respectively transfected and cotransfected into cultured HSC-T6 cells; expression of TGF betav1, TGF beta 3, type I collagen mRNA were detected by real-time quantitative PCR, expression of TGF beta 1 and type I collagen protein were detected by Western blot. The recombinant expression plasmid pcDNA3.1(+)-TGF beta 1 was transfected into cultured HSC-T6 cells; positive clones were selected by G418. The positive clones were transfected by the recombinant expression plasmid pcDNA3.1(+)-TGF beta 3; expression of TGF beta 1, TGF beta 3 and type I collagen mRNA were detected by real-time quantitative PCR; expression of TGF beta 1 and type I collagen protein were detected by Western blot.</p><p><b>RESULTS</b>HSC-T6 was transfected by recombinant expression plasmid pcDNA3.1(+)-TGF beta 1 and pcDNA3.1(+)-TGF beta 3 and the transfection efficiency was 28.2%. After the cells were transfected with pcDNA3.1-TGF beta 3, type I collagen mRNA and the protein expression in the cells were higher than those in the untransfected cells (control group) (P < 0.05). The increase reached to the maximal at 72 h after the transfection. Expressions of type I collagen mRNA and the protein in the cells transfected by pcDNA3.1(+)-TGF beta 1 were higher than in those cotransfected by pcDNA3.1(+)-TGF beta 1 and pcDNA3.1(+)-TGF beta 3 (P < 0.05). TGF beta 1 protein, type I collagen mRNA and type I collagen protein expression significantly decreased in the clones transfected by recombinant expression plasmid pcDNA3.1(+)-TGF beta 3 (P < 0.05), but the changes of TGF beta 1 mRNA were not significant (P > 0.05).</p><p><b>CONCLUSIONS</b>Expression of type I collagen increased after the cultured HSC-T6 cells were transfected by TGF beta 3 gene. The significant decrease of the expression of type I collagen of the TGF beta 3 gene transfected positive clones suggests that TGF beta 3 could inhibit the occurrence of liver fibrosis.</p>

Analysis of change in viral titers under different conditions in cultured cells persistently-infected with Japanese encephalitis virus / 中华实验和临床病毒学杂志

<p><b>OBJECTIVE</b>To investigate the change of viral titers under different conditions in cultured cells persistently-infected with different strains of Japanese encephalitis virus (JEV) and find out the factors that influence viral multiplication.</p><p><b>METHODS</b>JEV JaGAr-01 and Nakayama wild strains were used to infect human hepatoma cell line KN73 respectively, and the persistent infection model was established. Viral titers were examined by plaque methods using BHK cells. Human nerve fibroblastoma cell line IMR-32 was infected with the strains of the virus that can cause persistent infection, and the thermal sensitivity of the viral strains was measured at 30 degrees C and 37 degrees C. KN73 cells persistently infected with JEV were infected with two JEV strains respectively, and viral superinfection was studied. To explore the replication of the persistently-infected viruses, KN73 and IMR-32 cells were infected with the viral strains.</p><p><b>RESULTS</b>Two persistently infected viral strains did not show any thermal difference. The results of superinfection were that the viral titers of JaGAr-01 strains were 1.3 and 8.8 percent of the control, respectively, and the viral titers of Nakayama strain were 80 and 1.7 percent of the control, respectively. JaGAr-01 wild strains, Nakayama wild strains and their persistently-infected strains infected KN73 and IMR-32 respectively. The replication of the persistently-infected strains was obviously lower than the wild strains in KN73 cells, however, in IMR-32 cells their replication was similar.</p><p><b>CONCLUSIONS</b>The two strains of JEV were not found to be temperature-mutant. It is possible that mutant viruses containing DI particles exist in JEV persistently-infected strains. In different cells there may be different host factors hindering the replication of the persistently-infected strains.</p>

Changes of mast cells and protease activated receptor-2 in experimental rat liver fibrosis / 中华肝脏病杂志

<p><b>OBJECTIVE</b>To explore the quantity of mast cells and the role of protease activated receptor-2 (PAR-2) in experimental rat liver fibrosis.</p><p><b>METHODS</b>Rats were sacrificed at 0, 2, 4, 8, and 12 weeks after subcutaneous injection of CCl(4). Mast cells were displayed by toluidine blue stain. The content of liver hydroxyproline was measured by the method of base hydrolyzate. The mRNA expression and the protein expression of PAR-2 in livers were detected by RT-PCR and immunohistochemistry at each time point.</p><p><b>RESULTS</b>In normal rat livers there were a few mast cells (2.5+/-1.0) distributed along the hepatic portal areas. In the cirrhosis model group the number of mast cells in the livers increased degree by degree (2 weeks vs 4 weeks vs 8 weeks, 9.1+/-0.5 vs 15.7+/-3.0 vs 32.0+/-3.3; P less than 0.05), and they were distributed densely around the hepatic portal areas and the central veins. The content of liver hydroxyproline increased progressively from 0 to 12 weeks. In normal livers PAR-2 mRNA was hardly detected, at 2 weeks there was some expression of PAR-2 mRNA (PAR-2/beta-actin 0.15+/-0.01, P less than 0.05), at 4 weeks its expression increased (PAR-2/beta-actin 0.35+/-0.02, P less than 0.05) and it maintained a higher level (PAR-2/beta-actin 0.80+/-0.02, P less than 0.05) since then. The changing trend of the protein expression of PAR-2 was the same as that of PAR-2 mRNA expression.</p><p><b>CONCLUSIONS</b>PAR-2 mRNA expression and the protein expression of PAR-2 were consistent with the increase of the mast cells, and the content of liver hydroxyproline may play an important role in mediating liver fibrosis.</p>