By reviewing the practice of lab administrating system of reform in the School of Chinese Pharmaceutical Science in Guangzhou University of Chinese Medicine in recent years,we think that the setup of administrating system,the running mechanism and the reasonable allocation will be beneficial to and effective for further speeding up the experiment teaching reform in pharmacy specialty and improving the level of experiment teaching.

OBJECTIVE: The optimal multiplicity of infection (MOI) of the recombinant adenovirus Ad-Rad50-GFP carrying a mutant Rad50 gene expression region on the cell growth of nasopharyngeal carcinoma and the viral amplification efficiency of CNE1 cell infected by this adenovirus were studied. METHOD: The biological titer of Ad-Rad50-GFP was measured by end point dilution method. The impact of recombinant adenoviral vector transfection on the growth of CNE1 cells was observed by cell growth curve. Transfection efficacy of recombinant adenoviral vector was observed and calculated through fluorescence microscope. The expression f mutant Rad50 in the Ad-Rad50-GFP transfected CNE1 cells with optimal MOI was detected by Western Blot after transfection. RESULT: The biological titer of Ad-Rad50-GFP was 1.26 x 10¹¹ pfu/ml. CNE1 cell growth was not influenced significantly as they were transfected by recombinant adenoviral vector with MOI less than 50. Transfection efficacy of recombinant adenoviral vector was most salient at 24 hours after transfection, with the high expression of mutant Rad50, and the efficiency still remained about 70% after 72 hours. CONCLUSION Recombinant adenoviral vector Ad-Rad50-GFP could transfect CNE1 cells as well as result in the expression of mutant Rad50 in CNE1 cells effectively. MOI = 50 was the optimal multiplicity of infection of CNE1 cells transfected by recombinant adenoviral vector Ad-Rad50-GFP.
Adenoviridae ; Carcinoma ; Cell Line, Tumor ; Genetic Vectors ; Humans ; Nasopharyngeal Carcinoma ; Nasopharyngeal Neoplasms ; Transfection

Objective To investigate the modulatory effects of glycyrrhizin (GL) on the expressions of nu-clear factor kappa B (NF-κB), glucocorticoid receptor (GR) and cytokines in serum, and to explore the protective mechanism of GL in acute lung injury (ALI) induced by lipopolysaecharide (LPS). Method Twenty-four male Sprague-Dawley rats were randomly(random number) divided into three groups (n = 8 in each) : normal group, ALI group and GL treatment group. Rats in the ALI group and GL treatment group were administered with LPS (5 mg/kg) intravenously. In GL treatment group, rats were treated with GL (20 mg/kg) one hour before LPS injec-tion. The animals were sacrificed 4 hours after injection of LPS, and then the lung wewt/dry ratio and PaO_2 were measured, the histopathology of lung injury was observed under light microscope, the expressions of NF-κB mRNA and GR mRNA in lung tissues were detected by using RT-PCR, and the levels of NF-κB protein and GR protein were determined by using Western blot. The levels of TNF-α and IL-10 in serum were observed by using ELISA.Data were analyzed with SPSS version 13.0 software, and means were compared with analysis of variance and Stu-dent-Newman-Keuls test. Results (1) TNF-α levels in normal group, ALI group and GL treatment group were (43.96±7.57), (153.68±20.42), and (87.23±7.52) ng/L, respectively, and IL-10 levels were (24.72±8.03), (42.48±6.81) and (58.33±9.62) ng/L, respectively (F = 183.70, all P <0.01). (2) NF-κB mRNA expressions in normal group, ALI group and GL treatment group were (0.432±0.085), (3.414±0.521) and (1.894±0.272), respectively, and NF-κB protein levels were (45.6±7.3), (254.7±16.4)and (133.5 ±11.7) ng/L, respectively, and comparison between groups showed statistical significant (F = 187.82 and 1466.53, ALL P < 0.01). GR mRNA expressions in normal group, ALI group and GL treatment group were (0.434±0.013), (0.152±0.025) and (0.308±0.033), respectively, and GR protein levels were(54.6±6.5), (11.5±2.3)and (28.2±5.6) ng/L, respectively (F = 246.00 and 260.92, all P < 0.01). (3) Com-pared with normal group, infiltration of PMNs, capillary congestion and swelling were found in ALI group, and treatment with GL could attenuate the lung injury. Conclusions Glycyrrhizin has a protective effects on rats with ALI induced by LPS maybe through down-regulating the expressions of NF-κB mRNA and TNF-α mRNA, and up-regulating the expression of GR mRNA and level of IL-10 protein.

Objective To explore the application of enhanced CT in the differential diagnosis and treatment of peritonsillar abscess (PTA) and intratonsillar abscess (ITA).Methods Thirty-eight in-patients with clinically suspected PTA from June 2011 to June 2013 were included in this study.All these patients underwent enhanced CT scan for the throat region.According to CT results,the location of abscess was determined,and the thickness of the posterior wall of abscess as well as its distance with the internal carotid artery was calculated.Incision and drainage were then guided with this information.Results Twentysix of the 38 patients (68.4％) met the diagnosis of PTA,demonstrating a hypodense collection with rim enhancement in the peritonsillar space,including 4 cases with multilocular abscess.Ten cases (26.3％) should actually be diagnosed as ITA,with a abscess collection located in the palatine tonsil tissue.Two cases (5.3％) were diagnosed as peritonsillar cellulitis (PTC),showing diffuse isodense lesion around the peritonsillar space.The 26 cases of PTA and 10 cases of ITA patients were all cured using incision and drainage under the precise guidance of CT,while the 2 cases of PTC only treated with medicine.The mean distance between the posterior wall of abscess and the carotid artery ((x) ± s) were (0.76 ± 0.34) cm and (0.90 ± 0.37) cm for the two entities respectively,with no significant difference (P ＞ 0.05).Conclusions Enhanced CT scan can clearly demonstrate the characters of PTA and ITA,and make identification.Moreover,it is helpful for the determination of therapy,improving the success rate of drainage and reducing the potential risk of large artery injury.

Objective To explore the effects of epidermal growth factor-like domain 7 (EGFL7) gene silencing on the proliferation and invasion ablity of laryngeal carcinoma cells.Methods A lentiviral vector expressing EGFL7 shRNA was constructed and transfected into human laryngeal carcinoma Hep-2 cells.The expressions of EGFL7 mRNA and protein were detected by Real-time PCR and Western blot,respectively.Cell proliferation was evaluated by CCK-8 assay,cell cycle and apoptosis were tested by flow cytometry,and cell invasion was detected by transwell invasion assay.Results The relative expression level s of EGFL7 mRNA and protein in EGFL7-SuRNA group were svgnificantly lower than control group (P ＜0.001).Western blot analysis proved that the relative expression of EGFL7 protein in NC group,Lenti-NCgroup and Lenti-EGFL7 group was (0.39 ±0.12),(0.36 ±0.14) and (0.07 ±0.04),respectively.EGFL7 expression in Lenri-EGFL7 group was significantly inhibited than NC group (P ＜ 0.001),which confirmed that the recombinant lentivirus was successfully transfected into Hep-2 cells.The proliferation of Hep-2 cells was significantly inhibited after transfection (P ＜ 0.01).Compared with the NC group and Lenti-NC group,the proportion of cells in S phase was significantly increased in Lenti-EGFL7 group (P ＜ 0.01),and the proportion in G1 phase was significantly decreased (P ＜ 0.05).Cell apoptosis assay showed that the apoptotic rate in Lenti-EGFL7 group (66.2 ± 1.28) ％ was significantly increased in NC group (6.09-±3.28)％ and Lenti-NC group (9.86 ±2.13)％.In Transwell invision assay,the mean number of cells coming through the Metrigel in Lenti-EGFL7 group was significantly decreased than in the NC group and Lenti-NC group (P ＜ 0.01).Conclusion The proliferation and invasion ablity of laryngeal carcinoma Hep-2 cells can be inhibited by siRNA mediated EGFL7 gene silencing.

Clinical metagenomic next-generation sequencing (mNGS) entails unbiased shotgun sequencing of all microbial and host nucleic acids present in a clinical sample. By analyzing the microbiota diversity, taxonomic, and phylogenetic relationships of clinical specimens, metagenomics related analysis provides an opportunity to investigate substantial biological significance of different microbes. According to the published paper, most studies on mNGS mainly focused on the clinical impact evaluation. However, the studies focused on the analytical performance validation of mNGS before clinical application were rare. Here, a scheme, included intended use, method establishment, assay validation and standard operating protocol, for the laboratory validation of clinical metagenomics sequencing assay was provided by summarizing experiences of clinical laboratory department of Peking Union Medical College Hospital protocol and relevant research. In this scheme, we discussed important topics of mNGS laboratory validation as below: specimen type and pathogen list, bioinformatics pipeline setup, dry lab standard preparation and performance validation, mNGS workflow setup, background nucleotide acid evaluation, wet lab standard preparation and performance validation.

OBJECTIVETo explore the effects of exogenous carbon monoxide-releasing molecules 2 (CORM-2) on LPS-induced abnormal activation of platelets in peripheral blood of healthy human donors and its possible molecular mechanism.

METHODSVenous blood samples were collected from a healthy volunteer, and platelet-rich plasma (PRP) from the blood were isolated by differential centrifugation. The PRP was subpackaged into siliconized test tubes and then divided into control group, LPS group, inactive CORM-2 (iCORM-2) group, 10 µmol/L CORM-2 group, and 50 µmol/L CORM-2 group according to the random number table, with 3 tubes in each group. The PRP in control group did not receive any treatment. The PRP in LPS group received LPS (20 mL, 10 µg/mL) stimulation, and the PRP in iCORM-2 group, 10 µmol/L CORM-2 group, and 50 µmol/L CORM-2 group underwent the same LPS stimulation and treatment of 50 µmol/L iCORM-2, 10 µmol/L CORM-2, and 50 µmol/L CORM-2, respectively, with the dosage of 20 mL. After being cultured for 30 min, the platelet adhesion rate was determined by glass bottle method, the number of platelet spreading on fibrinogen was determined with immunofluorescent method, and the platelet aggregation rate was measured by turbidimetric method. The platelet poor plasma (PPP) was prepared from PRP, the levels of ATP in PPP and platelets were determined by chemical fluorescein method. The expressions of platelet glycoprotein I bα (GPIbα) and GPVI were analyzed by flow cytometer. The expressions of glycogen synthase kinase 3β (GSK-3β) and phosphorylated GSK-3β were determined by Western blotting and immunoprecipitation, respectively. Measurement of the above indices was repeated for 3 times. Data were processed with one-way analysis of variance and SNK test.

RESULTSCompared with those in control group, the platelet adhesion rates, numbers of platelets spreading on fibrinogen, platelet aggregation rates, expressions of GPIbα and GPVI in PRP, levels of ATP in PPP in LPS and iCORM-2 groups were significantly increased, while levels of ATP in platelets were significantly decreased (with P values below 0.05). Compared with those in LPS group, the former 7 indices in iCORM-2 group showed no significant differences (with P values above 0.05), while the levels of ATP in platelets in the 10 µmol/L CORM-2 and 50 µmol/L CORM-2 groups were significantly increased, and the other 6 indices in 10 µmol/L CORM-2 and 50 µmol/L CORM-2 groups were significantly decreased (with P values below 0.05). The expression levels of GSK-3β of the platelets in PRP in control, LPS, iCORM-2, 10 µmol/L CORM-2, and 50 µmol/L CORM-2 groups were 0.550 ± 0.060, 1.437 ± 0.214, 1.210 ± 0.108, 0.720 ± 0.010, and 0.670 ± 0.010, respectively, and the expression levels of the phosphorylated GSK-3β of the platelets in PRP in the above 5 groups were 0.950 ± 0.070, 1.607 ± 0.121, 1.420 ± 0.040, 1.167 ± 0.015, and 0.513 ± 0.122, respectively. Compared with those in control group, both the expression levels of GSK-3β and phosphorylated GSK-3β of the platelets in PRP in LPS and iCORM-2 groups were significantly increased (with P values below 0.05). The expression levels of GSK-3β and phosphorylated GSK-3β of the platelets in PRP between LPS group and iCORM-2 group were similar (with P values above 0.05). The expression levels of GSK-3β and phosphorylated GSK-3β of the platelets in PRP in 10 µmol/L CORM-2 and 50 µmol/L CORM-2 groups were significantly decreased compared with those in LPS group (with P values below 0.05).

CONCLUSIONSLPS stimulation can abnormally activate the platelets in peripheral blood of healthy human, but the abnormal activation can be inhibited by CORM-2 intervention, and the mechanism of the latter may involve the phosphorylation of GSK-3β mediated by GP.

Blood Platelets ; drug effects ; metabolism ; Carbon Monoxide ; metabolism ; Glycogen Synthase Kinase 3 ; Glycogen Synthase Kinase 3 beta ; Humans ; Lipopolysaccharides ; pharmacology ; Organometallic Compounds ; pharmacology ; Phosphorylation ; drug effects ; Platelet Activation ; drug effects ; Platelet Aggregation ; drug effects ; Platelet-Rich Plasma

Objective: To explore the clinical manifestation and treatment strategy for descending necrotizing mediastinitis (DNM). Methods: A total of 27 cases diagnosed as DNM from January 2010 to August 2018 in the First People’s Hospital of Foshan were reviewed. There were 16 males and 11 females, age ranged from 16 to 84 years. The clinical data were collected. SPSS 16.0 software and chi square test were used for statistical analysis. Results: ALL 27 cases were diagnosed as DNM by contrast-enhanced CT scan of the neck and chest. Among the 27 cases, 13 cases resulted from peritonsillar abscess, 8 cases from esophageal foreign body perforation, 5 cases from parapharyngeal and retropharyngeal space abscess, and one case from infection of oral cavity. These 27 cases were divided into three subtypes according to the sites of mediastinitis, including 11 cases for typeⅠ, 5 cases for type ⅡA and 11 cases for type ⅡB. Of 27 cases, 20 cases underwent transcervical drainage for DNM, of which 5 cases with tracheotomy and 6 cases with thoracic drainage, and finally 19 of the 20 patients were cured, and one patient died of bacteremia; 7 cases refused to received surgery and were routinely treated with antibiotics, of which, one case was cured and 6 cases died. The curative rate in patients underwent surgery was significantly higher than that in patients treated with medication (χ²=13.638, P<0.001). Among the 20 cured cases, 4 cases were combined with diabetes mellitus and 6 cases with necrotizing fasciitis, while in the 7 died cases, 5 cases were combined with diabetes mellitus and 6 cases with necrotizing fasciitis. The comorbidity rates of diabetes mellitus (χ²=4.074, P=0.044) and necrotizing fasciitis (χ²=4.457, P=0.035) in died cases were significantly higher than those in cured cases. Conclusion DNM is a serious infection, with high mortality especially in patients with diabetes and necrotizing fasciitis. Timely cervical and chest enhanced CT scan play vital role in its diagnosis. DNM can be treated effectively with transcervical drainage.

Objective: To study the clinical outcome of transoral CO₂ laser surgery for glottic cancer involving the anterior commissure. Methods: Thirty-two cases of glottic cancer involving the anterior commissure treated by transoral CO₂ laser surgery between March 2009 and December 2013 were retrospectively reviewed. Among these cases, 27 were T1bM0M0, 5 were T2N0M0. All cases were followed-up for more than 3 years. Results: All the 32 cases were successfully treated. Perioperative complications included injuries in the soft palate mucosa(13/32, 40.63%), loose incisors(3/32, 9.38%) and subcutaneous emphysema in the neck(2/32, 6.25%). During the follow-up period, granulation was found in all cases. Three cases had local recurrence. Two patients treated by a secondary transoral CO₂ laser surgery and the other case had total laryngectomy, all three cases were followed up for 5 years without recurrence. Two cases had regional recurrence but no primary site recurrence. One patient was treated by neck dissection, and followed up for 5 years without recurrence. The other patient died of supraclavicular and mediastinal lymph node metastasis and lung metastasis 40 months after operation. The overall 5-year survival rate was 90.6%. There was no significant difference in survival rate between T1bN0M0(92.6%) and T2N0M0(80.0%) (Log Rank χ²=0.788, P=0.375). The overall 5-year local regional control rate was 84.4%. In T1bN0M0 lesions, the 5-year local regional control rate was 92.6%, which was significantly higher than that in T2N0M0 lesions(40.0%) (Log Rank χ²=9.504, P=0.002). Conclusion With appropriate surgical indication, detailed preoperative evaluation, good surgical skill, transoral CO₂ laser surgery may achieve satisfactory outcome in the treatment of glottic cancer involving the anterior commissure.

The aim of this study is to investigate the effect of fibreoptic endoscopic of sallowing （FEES） in the assessment of pharyngeal dysphagia in post-irradiated patients with nasopharyngeal carcinoma. Fifty-three NPC patients with post-irradiated underwent FEES and video fluoroscopy（VF）.The results were analyzed using the Bolus Residue Scale and Rosenbek's penetration aspiration scale. The agreement in the detection of penetration and aspiration between FEES and VF of liquid（κ=0.56, 95% 0.38-0.73） and porridge（κ=0.64, 95% 0.43-0.81） was "fair". The detection rates of penetration on FEES with liquid and porridge were 60% and 51%, the detection rates of aspiration on VF with liquid and porridge were 70% and 53%. There were no statistical differences. The agreement in the detection of pharyngeal residue between FEES and VF of liquid （κ=0.38, 95%0.12-0.62） and porridge （κ=0.66, 95% 0.44-0.86） was "fair". The detection rates of pharyngeal residue on FEES and VF with porridge were 43% and 45%, the difference was not statistically significant. The detection rates of pharyngeal residue on FEES and VF with liquid were 44% and 24%, and the difference was statistically significant. FEES is an effective and valuable tool for evaluating pharyngeal dysphagia in post-irradiated patients with nasopharyngeal carcinoma.