Objective To evaluate the effectiveness of several methods in eliminating Mycoplasma contamination in cell culture of human hepatoma-derived cell line C3A. Methods PCR was performed to detect the Mycoplasmas contamination in cell cultures. The contaminated samples were treated by ciprofloxa-cin, heating, Plasmocure or co-culturing with macrophages. Transmission electron microscope ( TEM) and Q-PCR were used to comparatively analyze the cell morphology and gene expression before and after Plas-mocure treatment. Results Plasmocure succeeded in eliminating Mycoplasma contamination, while cipro-floxacin showed temporary efficacy. Heating and co-culturing with macrophages failed to eliminate Mycoplas-ma contamination. No Mycoplasma contamination in the Plasmocure-treated group was observed under TEM and the expression of ALB, TF and CYP3A4 genes were higher than the genes expressed in the contaminated group (P<0. 01). Conclusion Plasmocure treatment was effective in eliminating Mycoplasma contamina-tion in cell culture. Moreover, the cell morphology and gene expression in Plasmocure-treated group were re-stored to normal.

Background Many methods are widely studied to improve the therapeutic effect of trabeculectomy for glaucoma,and the clinical effectiveness of the amniotic membrane application during trabeculectomy is one of the research hotspots.Objective This study was to evaluate the efficacy and safety between trabeculectomy with (TE-AMT) and without amniotic membrane transplantation (TE-noAMT) for treating glaucoma.Methods Articles published from 1965 to December 2010 were searched from The Cochrane Library,PubMed,EMBASE,CNKI,Chinese Biomedicine Database,internet by computer,and manual search then was performed according to the predetermined strategy.Randomized controlled trials (RCT) on TE-AMT and TE-noAMT were included.The quality of included articles were scored based on Jadad table,and the relative parameters and information were extracted,including author,design of the trials,country,number of eyes and patients,gender,follow-up duration,age,classification of glaucoma,baseline IOP and loss rate.The main analysis indicators were the percentage of IOP reduction,completely operative successful rate,qualified success rate and adverse events.The pooled estimates were carried out with RevMan version 5.0 software.Results Nineteen RCTs documents were reviewed by meta-analysis with the Jaded scores ≥3 in 2 papers and ＜3 in 17 papers.Total 977 eyes of 818 patients were included.The weighted mean differences (WMD) of the percentage of IOP from baseline were (WMD =8.47％,95％ CI:5.20-11.75) at 6 months and (WMD=9.37％,95％ CI:4.97-13.77) at 12 months postoperatively.Relative risk (RR) of complete success rate and qualified success rate at postoperative 6 months were (1.40,95％ CI:1.19-1.65) and (0.47,95％ CI:0.22-1.00),respectively.RR values of adverse event such as shallow anterior chamber,hyphema,hypotony and choroidal detachment,failed filtering blebs at 6,12 months were 0.51 (95 ％ CI:0.30-0.85),0.43 (95％ CI:0.20-0.92),0.51(95％CI:0.26-1.00) and 0.57(95％CI:0.14-2.31),0.31(95％CI:0.20-0.47),0.31 (95％ CI:0.17-0.55),respectively.Conclusions TE-AMT appears to have better efficacy in lowering IOP,increasing complete success rate and reducing adverse event I in comparison with TE-noAMT.

Objective To detect mucin(MUC1,MUC2,MUC5AC) expressions in endoscopic ultrasound guided fine needle aspiration(EUS-FNA) specimens of the pancreatic neoplasms lesions and evaluate their diagnostic value.Methods The expression profile of MUC1,MUC2,MUC5AC from 54 EUS-FNA samples of pancreatic occupying lesions were examined by immunohistochemistry with mono- clonal antibodies.The results were correlated to the eytohistologic and clinical diagnosis of pancreatic cancer.Results Thirty-eight of the 54 patients had pancreatic cancer,6 patients had benign tumor,10 patients had chronic pancreatitis.The sensitivites of cytology and histology in diagnosing pancreatic cancer were 31.6%and 47.4%,respectively.The prevalence of MUC1,MUC2 and MUC5AC expres- sion in EUS-FNA samples of pancreatic cancer were 81.6%(31/38),10.5%(4/38),84.2%(32/38), respectively,and those in benign pancreatic neoplasms were 25%(4/16),31.3%(5/16),43.8%(7/16),respectively.There was statistically significance in MUCland MUCSAC expression between the two groups(P

Objective To study the effect of three kinds of disinfection liquid,namely chlorine dioxide,silver ion and BIOCIDE,on sterilization of Legionella pneumophila.Methods The suspended quantified disinfection test was conducted by using 7 type of Legionella p neumophila,0.5 ml of bacterial suspension and 4.5 ml disinfection liquid were mixed for certain period, the normal saline was used as the control.1.0 ml of the above liquid or its dilution was incubated with BCYE for 72 h in CO_2 and 35 ℃,the colony count and sterilization rate were calculated.Results The sterilization rate of chlorine dioxide was 6.04%,7.87% and 18.18% at the sterilization time of 0.5,1 and 2.5 h respectively.The sterilization rate of silver ion was 52.61%,99.46% and 99.80% at the time of 0.5,1,1.5 and 2.0 h respectively.As for BIOCIDE,the sterilization rate was 25.28%,93.62%,96.60% and 99.81% at the time of 0.5,1,1.5 and 2 h respectively.Conclusion Compared with chlorine dioxide,silver ion and biological disinfectant has a better sterilization effect on Legionella pneumophila.

Objective To construct the eukaryotic expression plasmid of pEGFPC1uPAR gene and explore the effect on the proliferation and invasion ability of Pam 212 cells. Methods The human uPAR cDNA was cloned by PCR, and inserted into the eukaryotic expression plasmid pEGFPC1. After identification of sequencing, the reconstructive plasmid was transformed transiently into Pam 212 cells, then the cell growth and the invasion ability were evaluated. Results The reconstructive plasmid of pEGFPC1uPAR was validated by sequencing. The reconstructive plasmid can promote the growth of Pam 212 cells and enhance the invasion ability. Conclusion The pEGFPC1uPAR plasmid was constructed successfully and uPAR was confirmed to promote the growth and the invasion ability of Pam 212 cells, which lay the foundation for further studies of uPAR in vivo.

Abstract: Objective　China was certified by World Health Organization as a malaria-free country in 2021. Malaria has become a rare infectious disease, and preventing the re-transmission of imported malaria and reducing deaths are the main challenges facing China after elimination of malaria. To analyze and clarify the characteristics of imported malaria deaths, and to provide prevention and treatment recommendations for overseas workers and health care workers. Methods　The data of 17 imported malaria deaths in the analysis of malaria deaths from 2016 to 2020 by the National Severe Malaria Treatment Expert Group were collected, and the relevant clinical epidemiological data and disease course records were analyzed. Results　The 17 malaria deaths were all imported from Africa with Plasmodium falciparum infection (malarial cerebral type), with no obvious regularity in the month of onset. Among them, 16 were male patients, 5 cases with underlying diseases such as diabetes mellitus, and 10 patients were first diagnosed in a second-level or lower hospital. Excluding patients who died of respiratory cardiac arrest in ambulances, the mean time difference between first onset and malaria diagnosis in 16 patients was 6.8 days (median 5.5 days), and the mean time between first onset and antimalarial treatment was 7.4 days (median 6 days), the mean time difference from initial onset to death was 10.3 days (median 8.5 days). Excluding cases with onset abroad and unknown time of return, all 14 patients developed the disease within 30 days after returning to China. Conclusion　All the fatal cases were infected with Plasmodium falciparum imported from Africa. The patients' awareness of actively seeking medical treatment is weak, and the delay in seeking medical treatment caused by the insufficient diagnosis and treatment capacity of health institutions at the township level and below is the main reason for the deaths. It is recommended to strengthen the self-protection awareness of staff in malaria-endemic areas overseas and raise their awareness of malaria. For returnees from areas with high malaria risk, primary medical institutions should pay attention to the patient's travel history in Africa, improve the awareness of malaria diagnosis, malaria diagnosis and treatment capabilities.

Objective To observe whether the intravenously transplanted homologous bone marrow mesenchymal stem cells (MSCs) can migrate to the impaired pancreas tissue in the rats with acute necrotizing pancreatitis (ANP). Methods MSCs were isolated from bone marrow of male Sprague-Dawley (SD) rats and adhered to the culture plate in vitro. Female rats were divided randomly into 3 groups:normal transplantation group, ANP transplantation group, ANP group with 10 rats in each group. ANP was induced by intraperitoneal injections with L-arginine. Both transplantation group received MSCs infusion through tail vein. 72 h later, the rats were sacrificed, the pancreas, heart, liver and kidney tissues were harvested, and the morphological changes were examined and scored, the characteristics of migration of MSCs to pancreas were detected with the expression of sry gene of Y chromosome by using chromogenic in situ hybridization (CISH). Results Rapid proliferation occurred in isolated MSCs after culture for 3 ～ 5 days and colonies were formed. After 3 generations, CD29 + CD44 + CD45-cells accounted for over 95% of all the cells. There ware massive tissue necrosis, inflammatory cells infiltration in the pancreas of ANP group, the pathological score was 10.31 ±0. 85, which were significantly higher than that in ANP transplantation group (7.30 ±0.79, P ＜ 0.05). Sry gene could be detected in the pancreas, heart, liver and kidney tissues. In addition, scattered distributed sry positive cells were observed in the normally transplanted pancreatic tissue, lots of sry positive cells were observed in the ANP transplanted pancreatic tissue, and they were located in the most injured areas.Conclusions The inflammatory pancreatic tissue has the ability of recruiting MSCs in vivo, which can alleviate local inflammation.

Objective To detect the expressions of hTERT and MMP-7 mRNA in the peripheral blood of patients with colorectal cancer and their correlation,and discuss the clinical significance of peripheral blood micro-metastasis.Methods Total RNA was extracted from blood samples by TRIzol and was reverse transcribed into cDNA.The mRNA expressions of MMP-7 and hTERT were detected by real-time PCR.Results Positive mRNA expression of hTERT was found in 63 of 181 patients with colorectal cancer,while positive mRNA expression of MMP-7 was seen in 61 cases.The positive rates of hTERT and MMP-7 mRNA had no significant differences in the patients' sex and age (P ＞ 0.05).The positive rate of MMP-7 mRNA in the patients with poorly differentiated adenocarcinoma (43.9 ％) was higher than that in the patients with moderately and well differentiated adenocarcinoma (27.8 ％) (x2 =4.873,P ＜ 0.05).The positive rate of HTERT mRNA in the patients with stage Ⅲ-Ⅳ (42.5 ％) was higher than that in the patients with stage Ⅰ-Ⅱ (24.0 ％) (x2 =6.591,P ＜ 0.05).The positive rate of MMP-7 mRNA in the patients with stage Ⅲ-Ⅳ (39.6 ％) was higher than that in the patients with stage Ⅰ-1Ⅱ (25.3 ％) (x2 =4.014,P ＜ 0.05).The positive rate of hTERT mRNA in the patients with distant metastasis (53.3 ％) was higher than that in the patients without distant metastasis (28.7 ％) (x2 =9.059,P ＜ 0.05).The positive rate of MMP-7 mRNA in the patients with distant metastasis(46.7 ％) was higher than that in the patients without distant metastasis (29.4 ％) (x2 =4.506,P ＜ 0.05).The positive rate of hTERT mRNA in the patients with positive CEA expression (52.6 ％) was significantly higher than that in the patients with negative CEA expression (30.1％) (x2 =6.735,P ＜ 0.05).The positive rate of MMP-7mRNA in the patients with positive CEA expression (60.5 ％) was significantly higher than those in the patients with negative CEA expression (26.6 ％) (x2 =15.490,P ＜ 0.05).In the patients without distant metastasis,any of hTERT and MMP-7 mRNA expression was thought to be blood micro-metastasis.The recurrence and metastasis rates after 6 months in the patients with blood micrometastasis (36.58 ％) were higher than those in the patients without blood micro-metastasis (7.14 ％)(x2 =13.027,P ＜ 0.05).The recurrence and metastasis rates after 12-month follow-up in the patients with blood micro-metastasis (68.29 ％) were higher than those in the patients without blood micro-metastasis (12.5 ％) (x2 =31.948,P ＜ 0.01).The sensitivity of combined detection was 80.00 ％,and the specificity was 79.03 ％ after 12-month follow-up.Spearman correlation analysis revealed that significant positive correlation existed between hTERT and MMP-7mRNA expression (r =0.341,P =0.000).Conclusion Detection of hTERT and MMP-7 mRNA expression can be used as reliable markers to predict peripheral blood micrometastasis in colorectal cancer patients.

Objective To evaluate the validity and reliability of the Chinese version of Alcohol Withdrawal Scale (AWS). Methods Totally 175 patients diagnosed as alcohol dependence according to the criteria of ICD-10 were studied. Intraclass correlation coefficient (ICC) analysis was applied for examining interrater consistency and Cronbach' s α for internal consistency. Factor analysis was used to examine the construct validity. Correlation analysis between AWS and CGI,Revised Clinic Institute Alcohol Withdrawal Syndrome Scale(CIWA-Ar) were conducted to evaluate the criterion validity. Based on clinical criteria,ROC curve was calculated so as to test the discriminative ability and establish the cut-off point of the scale. Results ( 1 ) Reliability: ICC value was 0.93,and Cronbach's α was 0.83,which indicated good interrater and internal consistency. (2) Validity:the correlation coefficients of the two subscale with the total scale score were 0.78,0.83 respectively. The correlation coefficients between the subscale were 0. 81 and factor analysis revealed that each item of the scale had relatively high load on the primary factor (0.409 ～0.926). At the time of admission,the total score of the AWS was positively correlated with that of CGI ( r = 0.71, P < 0.05 ). The total score of the AWS also was positively correlated with that of CIWA-Ar ( r = 0. 86, P<0. 05). As treatment went on,total score of the AWS showed a downward trend,at the end of the first week,the total score of the AWS was positively correlated with that of CGI ( r = 0.62, P<0.05). (3)The cut-off point of AWS for mild alcohol withdrawal state was determined as ≥3. With this cut-off point,AWS had both high sensitivity (92.1% ) and specificity (73.5% ) ,and the area under curve (AUC) was 0. 91. The cut-off point of AWS for moderate withdrawal state was determined as ≥7, and the sensitivity and specificity of AWS were 94.3 % and 89.7 % respectively, with the AUC of 0.94. The cut-off point of AWS for severe withdrawal state was determined as ≥ 10. With this cut-off point AWS had both high sensitivity (94. 9% ) and high specificity (92.6% ) .with the AUC of 0.93. Conclusion AWS has good reliability and validity and can reflect the change of the disease and the efficacy of treatment.

Objective To investigate the effects of maternal long-term exposure to low-dose trichlorfon on the serum paraoxonase (PON) activity of pregnant mice and development of embryos. Methods Female ICR mice (n=120) were randomly divided into control group and trichlorfon groups of different doses,and were managed by intragastric injection with trichlorfon of 0,2,10 and 50mg/kg,respectively. All the mice were managed once a day for a consecutive of 27 days,and were subjected to mating. The pregnant mice were continued to be managed with trichlorfon for 3 days,and were sacrificed on day 3 of gestation. The serum PON and acetylcholinesterase (AchE) activities were detected,and the development of embryos was evaluated. Results The serum PON activity of 2,10 and 50mg/kg trichlorfon group were (14.15?1.22),(12.78?1.80) and (10.45?1.95)IU/mL,respectively,and that of 50mg/kg trichlorfon group was significantly lower than that of control group [(13.37?2.31)IU/mL] (P0.05),while the the percentage of abnormal embryos of 50mg/kg trichlorfon group had an increased tendency. Conclusion Long-term exposure to low-dose trichlorfon can inhibit serum PON and AchE activity in pregnant mice without obvious effect on the development of embryos.