Effects of the proliferation, migration and apoptosis of AHVAC - on gastric cancer MKN-28 cells / 中国临床药理学与治疗学

AIM: To investigate the effects of agkis-trodon halys venom anti-tumor component (AHVAC-) on the biological behavior of gastric cancer MKN-28 cells. METHODS: Gastric cancer MKN-28 cells were treated with the experimental concentrations (5, 10, 15 μg/mL) of AHAVC- for 24 h. Cell proliferation and toxicity assay (cell counting kit-8, CCK-8) was used to detect the inhibition rates of the cells in different concentrations of AHVAC-. The migration ability of the cells was evaluated by wound-healing and Transwell assay. The apoptosis were observed by laser confocal microscopy with annexin V-mCherry/DAPI double staining, and the apoptosis rates were analyzed by flow cytometry with annexin V-FITC/PI double fluorescence staining. The protein level of Caspease-3 was determined by Western blot. RESULTS: Compared with normal control group, the results of AHVAC- concentration groups showed that with the increase of AHVAC- concentration, the proliferative activity of MN-28 cells decreased gradually (P<0.01), the cell migration ability decreased gradually (P<0.01), and the cell apoptosis rate increased (P<0.05). The expression of apoptosis-related protein Caspease-3 was up-regulated (P<0.01). CONCLUSION: AHVAC- inhibits proliferation and migration of gastric cancer MSN-28 cells and induces apoptosis.

Correlation between the early dynamic changes of lymphocyte and severity of disease in coronavirus disease 2019 patients / 中华危重病急救医学

Objective:To explore the relationship between the dynamic changes of lymphocytes in the early stage (within 72 hours of admission) and the severity of disease in patients with coronavirus disease 2019 (COVID-19).Methods:A retrospective study was conducted. The clinical data of COVID-19 patients admitted in Wenzhou Central Hospital from January 17, 2020 to February 14, 2020 were collected and analyzed. According to whether there was lymphopenia on the first day of admission [lymphocyte count (LYM) < 0.8×10 9/L], whether the difference between LYM on the third day and the first day of admission (ΔLYM) was less than 0, the patients were divided into four groups: the first group was LYM normal on the first day of admission, ΔLYM ≥ 0; the second group was LYM normal on the first day of admission, ΔLYM < 0; the third group was lymphopenia on the first day of admission, ΔLYM ≥ 0; the fourth group was lymphopenia on the first day of admission, ΔLYM < 0. The study endpoint was the development of severe/critically ill patients within 30 days after admission. Severe/critical standard referred to classification of Diagnosis and treatment protocol for coronavirus disease 2019 (trial version 5, revised edition). The differences in general information, laboratory results, and probability of developing severe/critical were compared among the four groups. Cox regression analysis was used to analyze the correlation between the early dynamic changes of lymphocytes and the probability of severe illness; and the Kaplan-Meier survival curve was drawn to assess the probability of severe illness in patients with different LYM groups. Results:A total of 104 patients with COVID-19 were enrolled, and 21 patients developed to severe/critical cases within 30 days of onset (accounting for 20.2%; 17 severe cases and 4 critical cases). There were significant differences in age ( F = 5.061, P = 0.003), white blood cell count (WBC) on the first day of admission ( Z = 10.850, P = 0.013), C-reactive protein (CRP) on the first day of admission ( Z = -4.449, P < 0.001), LYM on the first day of admission ( Z = 43.132, P < 0.001), LYM on the third day of admission ( Z = 40.340, P < 0.001), and the occurrence of severe/critical illness ( χ2 = 18.645, P < 0.001) among the four groups. Patients in groups 3 and 4 were older; patients in group 3 had the lowest WBC and LYM on the first day of admission; patients in group 4 had the highest CRP on the first day of admission, the lowest LYM on the third day of admission, and high proportion of severe/critical cases. Regarding the probability of severe/critically ill patients within 30 days of admission, univariate Cox regression analysis showed that the probability of severe/critical patients in group 4 was 12.7 times higher than that in group 1 [hazard ratio ( HR) = 12.732, 95% confidence interval (95% CI) was 3.951-41.025, P < 0.001]; age, CRP, albumin (ALB) and lymphocyte grouping were included in multivariate Cox regression analysis, the probability of severe/critically ill patients in group 4 was 6.4 times that of group 1 ( HR = 6.398, 95% CI was 1.757-23.301, P = 0.005); however, there was no difference in the probability among the group 1, 2 and 3. Kaplan-Meier survival curve showed that the probability of severe/critically ill patients in group 4 was significantly higher than that in groups 1, 2 and 3 (Log-Rank test: χ2 = 42.617, P < 0.001). Conclusions:Early lymphocyte dynamics change is related to the severity of patients with COVID-19. Patients with low LYM on the first day and continued decrease within 72 hours of admission have a higher probability to develop into severe/critically cases.

Clinicopathologic analysis of 38 male breast cancers / 中国实用外科杂志

OBJECTIVE: To analyze the clinicopathologic characteristics and molecular subtype in male breast cancer(MBC). METHODS: The clinical features, histological features and molecular subtype of 38 cases of male breast cancers admitted from January 2013 to March 2019 in Clinical Pathology Diagnostic Center of Ningbo were analyzed retrospectively. RESULTS: Thirty-eight patients were diagnosed with MBC, accounting for 0.83% of all the breast cancer in the same term. The median age was 68.5(range from 24 to 88). Tumor were located in left side in 20 cases, right side in 18 cases. Twenty-nine cases were diagnosed as invasive ductal carcinoma, two cases of secretory breast cancer, one case of invasive solid papillary carcinoma, two cases of intraductal papillary carcinoma with microinvasive, and four cases of encapsulated papillary carcinoma(two of them with microinvasive). Sixteen patients had lymphatic metastasis when the tumor was diagnosed. Imunohistochemically, ER was positive in 36 cases, while PR was positive in 35 cases.Eighteen cases were Luminal A type, while sixteen were Luminal B type and two cases were basal-like subtype.CONCLUSION: MBC is a rare malignant neoplasm, and most patients are diagnosed older and at advanced clinical stage,what is more, there is more possibility of second non-breast primary cancer. All suggesting that early diagnosis,treatment and more intensive surveillance are important. MBC needs further study.

DNA barcoding identification of Dendrobium huoshanense and its adulterants / 中国中药杂志

This research preliminarily discusses the relations of Dendrobium system growth through chloroplast gene rbcL, matK and the nuclear genome ITS2. The DNA barcoding universal sequence for authentication of the Dendrobium medical plants was slected and the possibility concerning utilizing the DNA barcoding to distinguish the D. huoshanenseand its adulterants was analyzed. Using the universal primer pair of ITS2, rbcL and matK, series of extended sequencing in the Dendrobium were conducted. Meanwhile, considering the different index about amplification and sequencing success rate of each sequence, the intraspecific and interspecific aberrance, the employment of BioEdit and MEGA 5.0 software were applied to establish the systematic tree of the NJ molecular and evaluate the diversified authentication capability of various sequences. The consequence demonstrates that the sequence of ITS2 is not only the largest one both in the intraspecific and interspecific aberrance of the Dendrobium but also has obvious barcoding gap. Considering the few overlap between the intraspecific and interspecific aberrance and the highest percentage regarding the formation of unilateral branch in diverse Dendrobium which have different ITS2 sequences, it can differentiate the species of Dendrobium. Furthermore, due to the inferior success rate of the rbcL and thematK and the lower reliability of NJ systematic tree, the percentage of the unilateral species which are generated by the systematic tree of rbcL and matK sequences is deficient. Therefore, the sequence of ITS2 can serves as DNA barcoding to distinguish the D. huoshanense, the D. moniliform and the D. officinale.

Postoperative pulmonary infection in patients undergoing surgical operation at different surgical opportunities / 中国感染控制杂志

Objective To compare postoperative pulmonary infection in patients undergoing surgical operation at different surgical opportunities.Methods Patients who underwent surgical operation in a hospital from January to December 2014 were surveyed retrospectively,patients' data were reviewed,patients with postoperative pulmonary infection were compared.Results A total of 20 343 surgical patients were investigated,227(1.12％) had postoperative pulmonary infection.Postoperative pulmonary infection rate in patients undergoing emergency operation was higher than that of selective operation (5.13 ％ vs 0.70 ％,x2 =307.49,P＜0.001).Postoperative pulmonary infection rates in patients undergoing emergency neurosurgical,general surgical,and cardiothoracic surgical operation were all higher than selective operation (all P ＜ 0.001).Among patients with pulmonary infection following emergency operation,the proportion of those who aged ＜60 years,with preoperative irrational antimicrobial use,cardiovascular disease,hypertension,and tracheotomy were all higher than those who received selective operation (all P＜0.05);the proportion of patients with cancer is lower than that of elective surgery patients(P＜0.05).Conclusion Postoperative pulmonary infection rate in patients undergoing emergency operation is higher than selective operation,postoperative pulmonary infection rates in patients undergoing emergency neurosurgical,general surgical,and cardiothoracic surgical operation are high.Patients with cardiovascular disease and hypertension are the focus of protection;for elective surgery,cancer patients are the focus of protection.

Component I from Agkistrodon acutus venom induces apoptosis of K562/A02 cells by promoting caspase 3 expression / 中国实验血液学杂志

To investigate the effects of component I from Agkistrodon acutus venom (AAVC-I) on the biological features of chronic myeloid leukemia cells, K562/A02 leukemia cells were cultured in the presence of AAVC-I (6.25 - 100 µg/ml) and the proliferation status was assayed by CCK-8 method. Morphological changes were observed by inversed microscope after Giemsa and Hochest 33258 staining, and cell apoptosis was detected by flow cytometry. Caspase 3 activity was tested by using Chromogenic Activity Assay Kit. The results showed that AAVC-I inhibited the growth of K562/A02 cells in time- and concentration-dependant manners, and the IC(50) at 48 h was 30.988 µg/ml. Giemsa and Hochest 33258 staining showed the typical apoptotic features in K562/A02 cells after induction with AAVC-I for 48 h. Flow cytometric analysis revealed that the percentage of the apoptotic cells reached from 0.88 up to 53.66 as the treated concentration was elevated from 0 to 50 µg/ml. Compared with the control group, the expression of caspase 3 in the tested group was enhanced in a dose-dependent manner (P < 0.05). It is concluded that AAVC-I can effectively inhibit the growth and promote apoptosis of K562/A02 cells. Elevated expression of caspase-3 may be attributed to the apoptosis of K562/A02 cells.

Effects of microcystin exposure to open field behavior and cognitive function of rats / 中华行为医学与脑科学杂志

ObjectiveTo observe the potential effects of microcystin-LR (MC-LR) to open field behaviors and cognitive functions of rats after low-dose and 60 days exposure.Methods20 SD male rats (4 weeks old) were grouped into control ( double distilled water),0.2,1.0 and 5.0 μg/kg groups by random number table.Rats were exposed to microcystin-LR intragastrically every two days for 60 d.Open fields and Morris water maze were employed to observe the spontaneous behavior and spatial learning and memory capacity of rats.ResultsFor each group,the duration in the central area of open field were ( 8.86 ± 3.38 ) s,( 3.09 ± 1.22 ) s,( 1.68 ± 0.24) s,(1.22 ±0.93) s,frequencies in central area were (4.80 ± 1.16) times,(4.40 ± 1.91 )times,( 1.40 ±0.23)times,(3.80 ± 1.02 ) times,movement duration were ( 137.30 ± 5.25 ) s,( 122.97 ± 9.50) s,( 108.86 ± 6.38 ) s,( 129.35 ± 9.40) s,the escape latency were ( 23.17 ± 4.28 ) s,(77.00 ± 8.31 ) s,(59.00 ± 11.89) s,( 63.80 ±16.09) s,respectively.Compared to control,the duration in the central area of open field decreased and escape latency increased significantly (P ＜ 0.05 ) in all the MC-LR treated groups; and frequencies in central area and movement duration of 1.0 μg/kg treated rats decreased significantly.No significant differences were observed between expsoured group and control group in spacial probe tests(P＞0.05).ConclusionMC-LR exerts effects on the spontaneous behavior and spatial cognitive ability of rats.The further study on neurobehavior effects of MC-LR is necessary.

The value of flow cytometry for the differential diagnosis between refractory cytopenia with multiple dysplasia and aplastic anemia / 中华内科杂志

Objective To evaluate the value of flow cytometry ( FCM) for the differential diagnosis between myelodysplasia (MDS) subtype refractory cytopenia with multiple dysplasia (RCMD) and aplastic anemia (AA). Methods The flow cytometric data of bone marrow samples from 168 cases of RCMD and 77 cases of AA were analyzed retrospectively in blind, and its results were compared with gold standard to evaluate its diagnosis values. Results The specificity of abnormal of single immunophenotype in the surface of granulocytes and myeloblasts was high (range 75.3% -100% ) , but the sensitivity was very low (range 5.4%-50% ). In parallel tests, the sensitivity and specificity of the combination of CD+34 cells≥1% , myeloblasts ≥3% , abnormal expression of CD117 in granulocytes and loss of CD13 in myeloblasts or increased intensity of CD33 in granulocytes were higher than other combinations. The sensitivity and specificity of above combination were more than 62% and 92% , respectively. In the scoring method,different score was given to 8 markers according to different diagnostic value, which were CD+34 cells ≥1 % ,myeloblasts≥3% , abnormal expression of CD117 in granulocytes, loss of CD13 in myeloblasts, increased intensity of CD33 in granulocytes, loss of CD13 in granulocytes, loss of CD10 in granulocytes, and decreased SSC in granulocytes. The sensitivity and specificity were both high if we defined that the total score ≥1.5 was RCMD and the score < 1. 5 was AA. Conclusions The value of abnormal of single immunophenotype for differential diagnosis between RCMD and AA is low. Parallel tests can increase the diagnostic sensitivity obviously and not decrease the specificity. CD+34 cells≥1% , myeloblasts≥3% and abnormal expression of CO117 in granulocytes were the most important markers. The scoring method is precise to distinguish RCMD from AA.

Intrahepatic biliary papillomatosis:a report of 9 cases / 中华肝胆外科杂志

Objective Intrahepatic biliary papillomatosis(IBP)is a rare disease that is characterized by multiple numerous papillary adenomas in the intrahepatic biliary duct(IBD).The clinical features and outcome,however,are not well known.The clinicopathologic features,treatments and follow-up results were retrospectively analyzed in order to improve the efficiency of diagnosis and treatment for the disease.Methods Between August 2006 and October 2008,9 patients were diagnosed with IBP by histological findings at a tertiary referral center,Ningbo Medical Center(University of Ningbo,College of Medicine,Ningbo,China).The authors retrospectively reviewed the medical records to obtain clinical,radiological and pathologic data.The therapeutic results and follow-up data were also reviewed.Results The ratio of male to female was 2:7 and the middle age was 59 years.Repeated episodes of fever and right upper quadrant abdominal pain with or without jaundice were the common clinical manifestations.There were intrahepatic choledocholithiasis and/or history of previous biliary operation in 8 cases.The level of CA19-9 and CEA were almost normal.Imaging workup demonstrated cyst-like dilatation of intrahepatic biliary tree with or without liver atrophy,which were more obvious in the mucin-hypersecreting IBP.All of the 9 cases underwent curative resection with an adequate resection margin.Macroscopic findings demonstrated that the tumors of 9 cases were all located in the left lobe with mucin-hypersecreting type in 7 cases.The diameter of numerous papillary granular was usually less than 10 mm(from 2 to 5 mm.frequently)and friable,that filled the dilated IHD dispersive.Fine fibrovascular cores lined by dysplastic epithelial cells were frequently found under microscope.Few foci of stromalinvasion were noticed in the two cases with malignant transformation.All of the 9 cases survived and there was no recurrence.Conclusion IBP occurs more often in middle and old women who have history of biliary disease. Repeated episodes of cholangitis are the common clinical manifestations. Extra- and intrahepatic biliary tract dilatation is the common imaging finding.MRCP/ERCP is more valuable than others in diagnosis. Clinical and histological finding shows that IBP is a premalignant disease with high malignant potential. Curative resection should be done as earlyas possible for the long-term survival rate. The use of cholangioscopy in operation and rapid biopsy of resection margin will benefit the curative resection.

Comparative study on the protective effects of Yinchenhao Decoction against liver injury induced by alpha-naphthylisothiocyanate and carbon tetrachloride / 中国结合医学杂志

<p><b>OBJECTIVE</b>To optimize the animal model of liver injury that can properly represent the pathological characteristics of dampness-heat jaundice syndrome of traditional Chinese medicine.</p><p><b>METHODS</b>The liver injury in the model rat was induced by alpha-naphthylisothiocyanate (ANIT) and carbon tetrachloride (CCl(4) ) respectively, and the effects of Yinchenhao Decoction (, YCHD), a proved effective Chinese medical formula for treating the dampness-heat jaundice syndrome in clinic, on the two liver injury models were evaluated by analyzing the serum level of alanine aminotransferase (ALT), asparate aminotransferase (AST), alkaline phosphatase (ALP), malondialchehyche (MDA), total bilirubin (T-BIL), superoxide dismutase (SOD), glutathione peroxidase (GSH-PX) as well as the ratio of liver weight to body weight. The experimental data were analyzed by principal component analytical method of pattern recognition.</p><p><b>RESULTS</b>The ratio of liver weight to body weight was significantly elevated in the ANIT and CCl(4) groups when compared with that in the normal control (P<0.01). The contents of ALT and T-BIL were significantly higher in the ANIT group than in the normal control (P<0.05,P<0.01), and the levels of AST, ALT and ALP were significantly elevated in CCl(4) group relative to those in the normal control P<0.01). In the YCHD group, the increase in AST, ALT and ALP levels was significantly reduced (P<0.05, P<0.01), but with no significant increase in serum T-BIL. In the CCl(4) intoxicated group, the MDA content was significantly increased and SOD, GSH-PX activities decreased significantly compared with those in the normal control group, respectively (P<0.01). The increase in MDA induced by CCl(4) was significantly reduced by YCHD P<0.05).</p><p><b>CONCLUSION</b>YCHD showed significant effects on preventing liver injury progression induced by CCl(4), and the closest or most suitable animal model for damp-heat jaundice syndrome may be the one induced by CCl(4).</p>

In vitro studies on asexual embryos and regenerated plantlets obtained from leaf organ of Panax notoginseng / 中国中药杂志

<p><b>OBJECTIVE</b>To study and improve the tissue culture technology of Panax notoginseng.</p><p><b>METHOD</b>Using the callus of leaf blade and leafstalk of P. notogingseng as explants, MS + 2, 4-D 1.5 mg x L(-1) as basal medium, the formation of asexual embryos was induced by added LFS, BA, KT or ZT 0.5 mg x L(-1), and cultured in dark. It cultured then in 2000 lx of illumination for 10-12 h x d(-1) to induce the asexual embryos germinating and developing to be the regenerated-plantlet.</p><p><b>RESULT AND CONCLUSION</b>Only the medium added with LFS could induce the formation of asexual embryos, and made it developed to be regenerated-plantlet. The inducing ratio of asexual embryos reached about 85%, and 30% of asexual embryos could grow and develop as robust regenerated-plantlets.</p>

Display of different injuries of recurrent laryngeal nerve in laryngeal electromyography / 中华耳鼻咽喉头颈外科杂志

<p><b>OBJECTIVE</b>To study the display of different types injuries of recurrent laryngeal nerve (RLN) in laryngeal electromyography (LEMG).</p><p><b>METHODS</b>LEMGs of one hundred and forty-seven patients (147 sides) with traumatic unilateral vocal cord paralysis (UVCP) were studied. After LEMGs, the RLNs exploration operations were performed. The condition of RLNs injury and laryngeal muscles was observed and recorded during the operation.</p><p><b>RESULTS</b>The severe injuries of RLNs were found during operation. The types of injuries were listed as ligation (58 cases), adhesion (28 cases) and cut (61 cases). The waveform morphology of LEMG was recorded less in the patients with the RLNs cut than that in the patients with the RLN ligation or adhesion, respectively. 75.4% RLNs cut showed spontaneous waveform while 96.4% RLNs adhesion and 94.8% RLNs ligation. When the RLN was cut off, single pattern was showed oftener. When the RLN was adhered or ligated, mixed pattern was showed oftener. 92.9% RLN adhesion showed misdirect-regeneration-potentials while 70.7% RLN ligation and 24.6% RLN cut. There were significant difference between two types, but the compound muscular active potential (CMAP) amplitude wasn't significantly different. Evoked amplitude could be recorded in 91.4% patients with ligation and its amplitude was (23.6 +/- 8.1)%, in 85.7% patients with adhesion and its amplitude (16.3 +/- 5.2)%, in 29.5% patients with cut and its amplitude (2.6 +/- 4.2)%.</p><p><b>CONCLUSIONS</b>The display of different injuries of RLN in LEMG presents significant difference. If RLN was cut off, the CMAP might be recorded in most cases. The clinical injury of RLN often is followed by sub-clinic reinnervation.</p>

Effects of Yiweining recipe on expressions of metalloproteinase-2 and cyclooxygenase-2 mRNAs in ectopic endometrium of rats with endometriosis / 中西医结合学报

OBJECTIVE: To explore the effects of Yiweining Recipe (YWNR), a compound Chinese herbal medicine, on expressions of metalloproteinase-2 (MMP-2) and cyclooxygenase-2 (COX-2) mRNAs in rats with endometriosis (EM). METHODS: Operational self-transplantation was applied in establishing the rat models. Detection of MMP-2 and COX-2 mRNAs was conducted with hybridization in situ. RESULTS: There were significant differences in the expressions of MMP-2 and COX-2 mRNAs between the untreated group and the high-dose YWNR-treated group. YWNR could reduce the expressions of MMP-2 and COX-2 mRNAs. CONCLUSION: YWNR can treat EM through reducing the positive expressions of MMP-2 and COX-2 mRNAs.

Fugene 6——a new approach to induce the gene transfection in high efficiency for eukaryotic cells in vitro / 免疫学杂志

Objective　To develop a new method to induce the gene transfection in high efficiency for eukaryotic cells in vitro. Methods　 Four kinds of p14ARF gene primarily deleted human carcinoma cell line including H460,A549,U251,and PC-3 were transfected with the human p14ARF expression vector (pCI-neo-p14ARF) by using the new nonliposomal transfection reagent Fugene 6. The efficiency of gene transfer was determined by screening the cells in G418. Results　 After 21 days' selection, G418-resistant clones were shown in all the transfected plate. PCR product of p14ARF gene was positive in all the G418-resistant clones. Cytotoxicity of Fugene 6 was detected. The cell proliferation activity was not affected when it was cultured in a high dose of Fugene 6. Conclusion　 These results demonstrate that Fugene 6 is a rapid, feasible, reproducible, and noncytotoxic gene transfection approach for eukaryotic expression vector in vitro.

Effect of Langchuangjing Granule on Expression of CD4~+CD8~+ and CD54 in Peripheral Blood of Lupus Mice / 中药新药与临床药理

Objective To study the immunoregulatory mechanism of Langchuangjing Granule(LG)in different dosages in lupus mice.Methods Three -month female NZB /NZWF1mice were treated with LG in three differen t dosages for six weeks and then the effect of LG on t he expression of CD4 + ,CD8 + and CD54in peripheral blood was obse rved.Re-sults LG in different dosages showed a tend ency of increasing the content of CD4 + and CD8 + ,decreased the serum con-tent of CD54and inhibited the hyper -expressions of CD54on the surface of lymphocyte and monocyte in plasma.T he above effects were stronger in middl e -and high -dosage of LG.Conclusion LG can improve the immune function an d restrain the excessive immunoreaction of lupus mice.