Metabolic stability and metabolic enzyme phaenotypes of lanceolatin B in liver microsomes of different species by UPLC-MS/MS / 中国中药杂志

To investigate the metabolic stability and parameters in vitro of lanceolatin B in liver microsomes of rats, human, Beagle dogs, and monkeys, and to identify the phaenotypes of CYP enzymes of lanceolatin B by using the liver microsome incubation system in vitro. After incubated with different species of liver microsomes, lanceolatin B was quantified by UPLC-MS/MS method to evaluate its metabolic stability and metabolic kinetic parameters in vitro. Lanceolatin B was incubated with specific inhibitors of CYP450 isoforms (CYP2E1, 2C19, 1A2, 2D6, 2C9, 3A4, and 2A1) to determine the phaenotypes of metabolic enzymes. The results showed that lanceolatin B was metabolized in the liver microsomes of rats and monkeys but not in the human and Beagle dogs. Their in vitro half-life T1/2 and intrinsic clearance rate CLint in rat and monkey liver microsomes were 11.57,8.07 min, and 0.12,0.17 mL•min⁻¹•mg⁻¹ without significant difference. The results of metabolic phenotyping indicated that CYP1A2 was mainly involved in the metabolism of lanceolatin B. There existed a difference in the metabolism of lanceolatin B in different types of liver microsomes. Several of CYP450 isoforms metabolized lanceolatin B together in liver microsomes of rats, in which CYP1A2 was the major enzyme mainly responsible for the metabolism of lanceolatin B.

In vitro metabolism of anti-tumor compound E7 in different species of liver microsomes / 中国中药杂志

To investigate the metabolic stability of E7 in liver microsomes of human, Beagle dog, Cynomolgus monkey and SD rats, and compare the metabolic differences between different species. Selective chemical inhibitors were used to determine the effects of different inhibitors on E7 metabolic rate, and predict the main enzymes involved in E7 metabolism in rat liver microsomes. The experimental results showed that the in vitro half-lives (T1/2) of E7 in liver microsomes of human, dog, monkey and rats were 57.75, 69.30, 16.90,30.13 min respectively. Their intrinsic clearance rate was 0.004 8, 0.004 0, 0.016 4 and 0.009 2 mL•min⁻¹•mg⁻¹ respectively. Hence, it could be speculated that the metabolic rate of E7 was similarly slow in human and dog liver microsomes; while it was similarly fast in monkey and rat liver microsomes. There was significant difference in metabolic rate of E7 between different species. The results showed that CYP2E1, CYP2A6, CYP1A2 and CYP2D6 might participate in metabolism of E7, while the contribution of polymorphic CYP3A4 was small.

High-frequency ultrasonography for diagnosis and differential diagnosis of acute scrotum in children / 中华男科学杂志

<p><b>Objective</b>To analyze the high-frequency ultrasound image features of acute scrotum in children and explore the value of high-frequency ultrasonography in the diagnosis and differential diagnosis of the disease.</p><p><b>METHODS</b>This retrospective study included 256 children aged 2 days to 14 years undergoing color Doppler ultrasonography at 2 hours to 3 days after onset of acute scrotum. We analyzed the morphology, internal echo and blood supply of the testis in comparison with the clinical and pathological results.</p><p><b>RESULTS</b>Among the 256 cases, acute testicular torsion was found in 23, of which 16 were treated by complete resection the necrotic testis and the other 7 by surgical reduction of testicular torsion. Ultrasonographically, the involved testes presented different degrees of increase or decrease in volume, with uneven internal echoes, irregular hypoechoic flakes, and testicular hydrocele. Color Doppler flow imaging (CDFI) showed significant blood flow signals around the diseased testes but none within them. Acute testicular appendix torsion was found in 116 cases, in which ultrasonography manifested nodules with round or oval abnormal echoes between the upper pole of the testis and caput epididymidis, first hypoechoic and then gradually increased, heterogeneous internally. CDFI revealed enlarged epididymides and enriched testicular blood flow but no blood flow signals in the nodules. The 103 cases of acute epididymitis were ultrasonographically characterized by varied degrees of swelling of the involved epididymis with uneven internal echoes and rich blood flow signals on CDFI. Six of the cases were diagnosed as acute orchitis, with the ultrasonographic features of testicular swelling and low but uniform internal echoes, with rich blood flow signals on CDFI. Incarcerated inguinal hernia was confirmed in 15 cases, in which ultrasonography revealed intrusion of the hernia into the obviously enlarged scrotal sac with the mesentery and intestine in it, and blood flow visible on CDFI. Acute scrotal wall hematoma and edema was found in 8 cases, with the ultrasonographic characteristics of scrotal wall thickening, with visible blood flow signals on CDFI.</p><p><b>CONCLUSIONS</b>High-frequency ultrasonography has a high sensitivity and specificity for acute scrotum in children, which can be applied as the first-choice clinical imaging modality and provide reliable evidence for the diagnosis and differential diagnosis of the disease.</p>

Therapeutic effect of recombinant tissue plasminogen activator on acute cerebral infarction at different times / 世界急诊医学杂志（英文）

BACKGROUND:The study aimed to compare the therapeutic effect of recombinant tissue plasminogen activator (rt-PA) on the onset of acute cerebral infarction (ACI) at different time points of the first 6 hours.METHODS:A retrospective analysis was conducted in 74 patients who received rt-PA thrombolysis treatment within 4.5 hours after ACI and another 15 patients who received rt-PA thrombolysis treatment between 4.5-6 hours after ACI.RESULTS:National Institute of Health Stroke Scale (NIHSS) scores were statistically decreased in both groups (P>0.05) at 24 hours and 7 days after ACI. There was no significant difference in modified ranking scores and mortality at 90 days after the treatment between the two groups (P>0.05).CONCLUSIONS:The therapeutic effect and mortality of rt-PA treatment in patients with ACI between 4.5-6 hours after the onset of the disease were similar to those in patients who received rt-PA within 4.5 hours after the onset of this disease. Therefore, intravenous thrombolytic therapy for ACI within 4.5-6 hours after ACI was effective and safe.

Expression of alternatively spliced human tissue factor in acute leukemia cells / 中国实验血液学杂志

The high expression of tissue factor (TF) is related to the coagulation disorder in acute leukemia. TF in blood circulation is mainly expressed in cells, microparticles (MP) and alternatively spliced human tissue factor (asHTF). To elucidate the role of TF in the coagulation disorder of acute myeloid leukemia (AML), RT-PCR was performed on 6 common AML cell lines NB4, HL-60, Kasumi-1, U937, K562 and THP-1. The results showed that only NB4 and U937 cells expressed baseline full-length TF and asHTF which were proved by sequencing. The flow cytometric detection, TF activity and TF antigen tests in NB4 and U937 cells revealed that the asHTF was expressed in trace amount and almost had no activity, while the TF antigen and activity in microparticles were significantly higher than that in asHTF. It is concluded that asHTF may play an unimportant role in the coagulation disorder of AML. Microparticle associated tissue factor (MP-TF) is the predominant source of TF activity released from AML cells.

Direct ambulance transport to catheterization laboratory reduces door-to-balloon time in patients with acute ST-segment elevation myocardial infarction undergoing primary percutaneous coronary intervention: the DIRECT-STEMI study / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Primary percutaneous coronary intervention (PCI) has been clearly identified as the first therapeutic option for patients with acute ST-segment elevation myocardial infarction (STEMI). The importance of reducing door-to-balloon (D2B) time has gained increased recognition. This study aimed to assess the feasibility, safety and efficacy of the strategy of direct ambulance transportation of patients with acute STEMI to catheterization lab to receive primary PCI.</p><p><b>METHODS</b>The study population included 141 consecutive patients with chest pain and ST-segment elevation who were admitted to the catheterization laboratory directly by the ambulance and underwent primary PCI (DIRECT group). Another 145 patients with STEMI randomly selected from the PCI database, were served as control group (conventional group); they were transported to catheterization laboratory from emergency room (ER). The primary endpoint of D2B time, and secondary endpoint of in-hospital and 30-day major adverse cardiac events (MACE, including death, non-fatal reinfarction, and target vessel revascularization) were compared.</p><p><b>RESULTS</b>Baseline and procedural characteristics between the two groups were comparable, except more patients in the DIRECT group presented TIMI 0-1 flow in culprit vessel at initial angiogram (80.1% and 73.8%, P = 0.04). Comparing to conventional group, the primary endpoint of D2B time was reduced ((54 ± 18) minutes and (112 ± 55) minutes, P < 0.0001) and the percentage of patients with D2B < 90 minutes was increased in the DIRECT group (96.9% and 27.0%, P < 0.0001). The success rate of primary PCI with stent implantation with final Thrombolysis in Myocardial Infarction (TIMI) 3 flow was significantly higher in the DIRECT group (93.8% and 85.2%, P = 0.03). Although no significant difference was found at 30-day MACE free survival rate between the two groups (95.0% and 89.0%, P = 0.06), a trend in improving survival status in the DIRECT group was demonstrated by Kaplan-Meier analysis.</p><p><b>CONCLUSION</b>Direct ambulance transport of STEMI patients to the catheterization laboratory could significantly reduce D2B time and improve success rate of primary PCI and 30-day clinical outcomes.</p>

Expression of β-1,4-galactosyltransferase I in a surgically-induced rat model of knee osteoarthritic synovitis / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>There are few reports of a biological role for glycosyltransferases in the infiltration of osteoarthritic synovitis. The aim of this research was to investigate the expression and cellular location of β-1,4-galactosyltransferase I (β-1,4-GalT-I) in a surgically-induced rat model of knee osteoarthritis (OA), and explore the role of β-1,4-GalT-I in the pathogenesis of OA.</p><p><b>METHODS</b>Male Sprague-Dawley rats were randomly divided into three groups: OA group, sham group and normal group. The model of OA was established in the right knees of rats by anterior cruciate ligament transaction (ACLT) with partial medial meniscectomy. Fibroblast-like synoviocytes (FLSs) obtained from normal rat synovial tissue were cultured. The expression of β-1,4-GalT-I mRNA in the synovial tissue, articular cartilage and FLSs treated with tumor necrosis factor-α (TNF-α) were assayed by real-time PCR. Western-blotting and immunohistochemisty were used to observe the expression of β-1,4-GalT-I at the protein level. Double immunofluorescent staining was used to define the location of the β-1,4-GalT-I with macrophage-like synoviocytes, FLSs, neutrophils, and TNF-α in the OA synovium. The alteration of TNF-α in FLSs which were treated with lipopolysaccharide (LPS) and β-1,4-GalT-I-Ab were detected by enzyme-linked immunosorbent assay (ELISA).</p><p><b>RESULTS</b>The mRNA and protein expression of β-1,4-GalT-I increased in synovial tissue of the OA group compared with the normal and sham groups at two and four weeks after the surgery, however, no significant difference appeared in the articular cartilage. Immunohistochemistry also indicated that the β-1,4-GalT-I expression in OA synovium at four weeks after surgery increased sharply compared with the control group. β-1,4-GalT-I co-localized with macrophage-like synoviocytes, FLSs, neutrophils and TNF-α in rat OA synovitis. Moreover, in vitro β-1,4-GalT-I mRNA in FLSs was affected in a dose- and time-dependent manner in response to TNF-α stimulation. ELISA revealed that the expression of TNF-α was attenuated in FLSs in vitro when treated with anti β-1,4-GalT-I antibody.</p><p><b>CONCLUSION</b>β-1,4-GalT-I may play an important role in the inflammation process of rat OA synovial tissue which would provide the foundation for further researching into the concrete mechanism of β-1,4-GalT-I in OA synovitis.</p>

Acupuncture treatment of regulating spirit, activating blood and relieving pain for thalamic pain / 中国针灸

<p><b>OBJECTIVE</b>To compare the clinical effect of acupuncture treatment and western medicine Carbamazepine for thalamic pain.</p><p><b>METHODS</b>Crossover trial design was used, 11 cases diagnosed as thalamic pain were randomly divided into two groups according to the mini-unbalance-index method, group I (with 6 cases received acupuncture first and then western medicine) and group II (with 5 cases received western medicine first and then acupuncture). When the effects were evaluated, the two groups were named as acupuncture group and western medicine group, 11 cases in each group. The method of clearing away the heart fire, regulating the spirit, activating blood and relieving pain was adopted in acupuncture treatment, Ximen (PC 4), Yinxi (HT 6), Xuehai (SP 10) and Zhaohai (KI 6) were selected; the western medicine group was treated with oral administration of Carbamazepine, and one course as well as the eluting period were both 10 days. The effects were evaluated with visual analogue scale (VAS) and evaluation scale of Anderson Cancer Center pain in US (MD Pain Evaluation value) respectively.</p><p><b>RESULTS</b>The VAS and MD value in two groups were obviously decreased after treatment (both P < 0.05), while there was no significant difference between two groups; the markedly effective rate of pain relieving in acupuncture group was 63.6% (7/11), which was higher than that of 36.4% (4/11) in western medicine group, but there was no significant difference between two groups.</p><p><b>CONCLUSION</b>Acupuncture treatment of regulating spirit, activating blood and relieving pain has a better therapeutic effect for thalamic pain, and can reach to the same therapeutic effect with western medicine Carbamazepine.</p>

Research on mechanism of chloroquine phosphate-induced U937 cell apoptosis / 上海第二医科大学学报

Objective To observe the effects of chloroquine phosphate on apoptosis of leukemic cell line U937, and investigate whether chloroquine phosphate induces leukemic cell apoptosis by normalizing protein PNAS-2's abnormal subcellular location. Methods Chloroquine phosphate of different concentrations were added into culture fluid of leukemic cell line U937 at logarithmic phase. MTr was used to measure cell proliferation, flow cytometry and laser confocal microscopy were applied to detect cell apoptosis, and immunofluorescence technology was employed to observe the effects of chloroquine phosphate on the changes of subcellular location of protein PNAS-2. Results Apoptosis of leukemic cell line U937 was significantly induced by 50 μg/mL chloroquine phosphate, and subcellular location of protein PNAS-2 was changed. Conclusion Chlorequine phosphate can induce apoptosis of leukemic cell line U937, and the mechanism may be related to the normalization of PNAS-2's abnormal subcellular location in U937 cell line. Chloroquine phosphate has the potential to be used in leukemic therapy.

Preparation and identification of monoclonal antibody against PNAS-2 protein / 中国实验血液学杂志

This study was purposed to prepare and primarily identify the specific monoclonal antibodies (McAbs) against the apoptosis related protein PNAS-2 so as to provide the essential tool for study of PNAS-2 function. The McAbs against PNAS-2 were prepared via the immunization of mice, cell fusion and cloning using synthetic peptide of PNAS-2 as immunogen; the specificity, titer and subtype of McAb were detected by Western blot, ELISA and immunofluorescence. The results showed that the stable hybridoma cell line S-31-7 producing McAbs against PNAS-2 protein was successfully obtained. The immunoglobulin of the McAb was identified to be IGg1lambda. The titer of ascetic fluid fled McAb were 1:8,000. A single specific band with 28 kD was shown in Western blot test, and the antigen recognized was present in cell cytoplasm by immunofluorescence. In conclusion, the obtained McAb against PNAS-2 displays strong specificity and high titer, which may be applied to the advanced research on PNAS-2 protein.

Polymorphisms of CYP3A5 gene in acute leukemia patients and their role in chemotherapy and prognosis / 中国实验血液学杂志

The objective of study was to investigate the role of the polymorphisms and protein expression of CYP3A5 gene in the therapy and prognosis of acute leukemia (AL) patients, the polymorphisms of CYP3A5 gene and the expression of protein CYP3A5 were detected by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) and immunohistochemistry method respectively. The results showed that there were three CYP3A5 genotypes in the 88 cases, namely CYP3A5*1/*1, CYP3A5*1/*3 and CYP3A5*3/*3 with frequencies of 26%, 50% and 24%, respectively. There were no significant differences in clinic data between the three groups, but the expressions of CYP3A5 of three groups were (36.6+/-19.2)%, (7.8+/-9.2)%, (0.5+/-0.9)%, the OS were (11.6+/-2.1) months, (30.5+/-12.2) months, (52.3+/-8.5) months, and the DFS were (7.5+/-1.8), (27+/-15.8), (52.3+/-8.1) months, respectively (p<0.05). It is concluded that the polymorphism of CYP3A5 gene is not related with the morbidity of AL, but closely associated with the expression of CYP3A5 in AL patients, and the latter are closely associated with the chemotherapeutic effect and prognosis. CYP3A5 genotype may be used as a new predictor to the chemotherapeutic effect and prognosis in AL cases.

Expression spectra of apoptosis-related gene pnas-2 / 中国实验血液学杂志

To explore the expression spectra of apoptosis-related gene pnas-2 in normal tissues and acute leukemia (AL) patient tissues, the expressions of pnas-2 gene in tissues including heart, brain, placenta, lung, liver, skeletal muscle, kidney, pancreas, spleen, lymph node, thymus, leukocyte, bone marrow and fetal liver were detected by Northern blot. The expressions of pnas-2 in samples including 44 de novo, 9 non-CR, 27 CR and 12 relapsed AL patients were measured by real-time RT-PCR and Northern blot, and the expression levels of pnas-2 in normal and tumor tissues from 31 patients with malignancies were also detected. The results showed that pnas-2 was not expressed in the most tissues except in placenta. The results of real-time PCR indicated that pnas-2 expressions in samples of de novo, non-CR and relapsed patients ware significantly higher than that in CR, tumor tissues and normal tissues. In serial monitoring of 7 AL patients, the expression level of pnas-2 was high at first visit examination, but remarkably decreased after remission, and the pnas-2 expression level increased again when relapsed. It is concluded that the pnas-2 is specifically up-regulated in acute leukemia patients, which might be an oncogene and participate in leukemogenesis.

Correlation between expression of apoptosis-related gene pnas-2 and leukemia / 中国实验血液学杂志

The study was purposed to explore the correlation between apoptosis-related gene pnas-2 and leukemia. The RT-PCR was performed to detect the expression levels of pnas-2 gene in NB4, K562, U937 cells before and after treatment with AS(4)S(4), and to analysis the expression change of pnas-2 gene in bone marrow cells from patients with acute leukemia before and after chemotherapy. The results showed that the expression of pnas-2 gene in arsenic sulfide treated NB4 cells was down regulated in time-dependent manner, but the same outcome in K562 and U937 cells after being treated with AS(4)S(4) was not found. The positive expression rate of pnas-2 in cells from untreated patients with acute leukemia was 100%, and was significantly higher than that in normal control group. After chemotherapy, the expression was negative in complete remission patients, whereas in no-remission patients there were no significant differences of expression of pnas-2 before and after treatment. It is concluded that the pnas-2 gene may be closely related with apotosis of arsenic sulfide treated APL cells, and may consider as a molecular biological remission marker in acute leukemia.

Effect of daunorubicin and cytarabine on cell line NB4 / 中华血液学杂志

<p><b>OBJECTIVE</b>To explore whether daunorubicin (DNR) combined with cytosine arabinoside (Ara-C) and DNR alone have similar effect on acute promyelocytic leukemia (APL) cell line NB4 and acute myeloblastic leukemia cell line HL-60 in vitro.</p><p><b>METHODS</b>Cell morphology, cells viability, and cell apoptosis (Annexin-V by flow cytometry assay) were analysed.</p><p><b>RESULTS</b>After incubation with DNR plus Ara-C for 24 hours,NB4 cell viability [(36.75 +/- 3.82)%] (n = 6) and cell apoptosis rate [(21.24 +/- 5.82)%] (n = 3) did not change significantly compared to that treated with DNR alone for 24 hours [(35.73 + 6.28 )%, (22.55 +/- 3.26)%, respectively] (P > 0.05). However, HL-60 cell viability [(67.17 +/- 2.07)%] and cell apoptosis rate [(48.05 +/- 0.92)%] changed significantly in DNR plus Ara-C group compared with DNR alone [(63.31 +/- 1.80)% ,(41.51 +/- 0.89)%, respectively] (P < 0.01 and < 0.05, respectively).</p><p><b>CONCLUSION</b>DNR plus Ara-C and DNR alone have similar effect on NB4 cells, but have different effect on HL-60 cells.</p>

Effects of quercetin on morphology and VEGF secretion of leukemia cells NB4 in vitro / 中华肿瘤杂志

<p><b>OBJECTIVE</b>To investigate the effects of quercetin on cell morphology and VEGF expression of acute myeloblastic leukemia cells NB4 in vitro.</p><p><b>METHODS</b>The cytomorphology of NB4 cells was assessed by Wright-stain, apoptosis rate by apoptotic marker Annexin V, and VEGF secretion level by ELISA.</p><p><b>RESULTS</b>Typical apoptosis was found in NB4 cells after treatment with quercetin. Apoptotic marker Annexin V analysis showed that the apoptotic rate of NB4 cells was increased after treatment with quercetin. The secretion of VEGF of NB4 cells was significantly decreased after treatment with quercetin.</p><p><b>CONCLUSION</b>Quercetin can induce apoptosis and inhibit secretion of VEGF in NB4 leukemia cells.</p>

Use of 16S-rRNA Hybridization Probes to Detect the Cellulolytic Bacteria in Rumen / 微生物学通报

In this study, the general bacterial probe and specific cellulolytic bacterial probes were used to quantify the bacteria in rumen. The total RNA were extracted and then hybridized with general bacterial probe after a dilution of concentration. The result showed that there was a high correlation between the hybridization signal and the dilution of total bacterial RNA. Based on the result above, the quantities of three cellulolytic bacteria in rumen sample were detected. The comparative RNA percentage of three cellulolytic bacteria to total bacterial RNA were similar to the previous reports. It can be concluded that the quantification of bacteria in rumen could be conducted by this approach, and which could be used in future research.