Effects of sFRP3 overexpression on the activation and proliferation of murine cardiac fibroblasts / 安徽医科大学学报

Objective @#To explore the role of secreted frizzled ⁃related protein 3 ( sFRP3) , a regulator of the Wnt signaling pathway , in the activation and proliferation of murine cardiac fibroblasts ( CFs ) . @*Methods @#Neonatalmice aged 1 -3 days were obtained for surgical procedures to collect heart tissues. After digestion, CFs were isola.ted and cultured. Transforming growth factor-beta 1 (TGF-β1 ) stimulation was used to induce activation and prolif.eration in CF's after they adhered to the culture dish. Once the model was confirmed, experimental and controlgroups were transfected with sFRP3 overexpression plasmids and empty plasmids for 24 -48 hours. Expression lev.els of sFRP3, Periostin (POSTN), Type I collagen ( Collagen I ), and proliferating cell nuclear antigen ( PCNA) were assessed at the molecular level using Westerm blot and qR'T-PCR. Changes in cell proliferation capacitywere examined using M'TT, CCK-8, and EdU staining methods.@*Results@#In the TGF-β1-induced activation andproliferation model of CFs, compared to the control group, the model group exhibited decreased expression of sFRP3 protein and mRNA , while the expression of activation and proliferation-related proteins PCNA, POSTN, andCollagen I was upregulated. Furthermore, in CFs overexpressing sFRP3 through plasmid transfection, the proteinand mRNA expression of PCNA, POSTN, and Collagen I decreased compared to the empty vector group. MTTCCK-8, and EdU experiments indicated a significant decrease in the proliferative activity of CFs in the sFRP3 over0verexpression of sFRP3 markedly inhibits theexpression group compared to the empty vector group. @*Conclusion@#Overexpression of sFRP3 markedly inhibits theactivation and proliferation of CFs, suggesting that sFRP3 may be a key gene involved in the regulation of CF activation and proliferation.

Effects of DNMT3A on mouse cardiac fibroblast proliferation and migration under high glucose environment / 中国药理学通报

Aim To investigate the effect of DNA methyltransferase 3A (DNMT3A) on the proliferation and migration of cardiac fibroblasts (CFs) in C57 mice under high glucose environment. Methods The hearts of C57 mice were taken from 1 to 3 days. After cutting and digesting, CFs were extracted by differential adherance centrifugattion and observed under microscope. After cell attachment, the cells were cultured under low glucose (5.5 mmol • L

Localized delivery of nanomedicine and antibodies for combating COVID-19

The severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) pandemic has been a major health burden in the world. So far, many strategies have been investigated to control the spread of COVID-19, including social distancing, disinfection protocols, vaccines, and antiviral treatments. Despite the significant achievement, due to the constantly emerging new variants, COVID-19 is still a great challenge to the global healthcare system. It is an urgent demand for the development of new therapeutics and technologies for containing the wild spread of SARS-CoV-2. Inhaled administration is useful for the treatment of lung and respiratory diseases, and enables the drugs to reach the site of action directly with benefits of decreased dose, improved safety, and enhanced patient compliance. Nanotechnology has been extensively applied in the prevention and treatment of COVID-19. In this review, the inhaled nanomedicines and antibodies, as well as intranasal nanodrugs, for the prevention and treatment of COVID-19 are summarized.

Erratum: Author correction to "Neutralization of SARS-CoV-2 pseudovirus using ACE2-engineered extracellular vesicles" Acta Pharmaceutica Sinica B 12 (2022) 1523-1533

[This corrects the article DOI: 10.1016/j.apsb.2021.09.004.].

Neutralization of SARS-CoV-2 pseudovirus using ACE2-engineered extracellular vesicles

The spread of coronavirus disease 2019 (COVID-19) throughout the world has resulted in stressful healthcare burdens and global health crises. Developing an effective measure to protect people from infection is an urgent need. The blockage of interaction between angiotensin-converting enzyme 2 (ACE2) and S protein is considered an essential target for anti-severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) drugs. A full-length ACE2 protein could be a potential drug to block early entry of SARS-CoV-2 into host cells. In this study, a therapeutic strategy was developed by using extracellular vesicles (EVs) with decoy receptor ACE2 for neutralization of SARS-CoV-2. The EVs embedded with engineered ACE2 (EVs-ACE2) were prepared; the EVs-ACE2 were derived from an engineered cell line with stable ACE2 expression. The potential effect of the EVs-ACE2 on anti-SARS-CoV-2 was demonstrated by both in vitro and in vivo neutralization experiments using the pseudovirus with the S protein (S-pseudovirus). EVs-ACE2 can inhibit the infection of S-pseudovirus in various cells, and importantly, the mice treated with intranasal administration of EVs-ACE2 can suppress the entry of S-pseudovirus into the mucosal epithelium. Therefore, the intranasal EVs-ACE2 could be a preventive medicine to protect from SARS-CoV-2 infection. This EVs-based strategy offers a potential route to COVID-19 drug development.

Inhaled heparin polysaccharide nanodecoy against SARS-CoV-2 and variants

The heparin polysaccharide nanoparticles block the interaction between heparan sulfate/S protein and inhibit the infection of both wild-type SARS-CoV-2 pseudovirus and the mutated strains through pulmonary delivery.Image 1.

Effects of ALKBH5 regulation on proliferation of cardiac fibroblasts in rats / 安徽医科大学学报

Objective @#To investigate the role of N-methyladenosine(m6 A) demethylase ALKBH5 in the prolifera- tion and activation of cardiac fibroblasts( CFs) in rats．@*Methods @#The CFs taken from SD rats in 1 to 3 days were isolated by differential adhesion and observed under microscope．After cells were adherently grown to appropriate density，the cells were induced by TGF-β1 for modeling．The model cells were divided into the overexpression of ALKBH5 group infected by lentivirus and the negative control group for 24－48 hours. RT-qPCR was used to detect mRNA expression of ALKBH5，α-smooth muscle actin( α-SMA) ，type I collagen ( Collagen Ⅰ ) and proliferating cell nuclear antigen (PCNA) ．The expression of ALKBH5、α-SMA、Collagen Ⅰ and PCNA were assayed by West- ern blot．The cell proliferation activity was tested by CCK-8 assay and EdU. @*Results @#Compared with the control group，the protein and mRNA of ALKBH5 were reduced in the model group active by TGF-β1．Meanwhile，the bi- omarkers of activation，such as PCNA，α-SMA and Collagen Ⅰ , increased significantly．Besides，the protein and mRNA of PCNA、α-SMA and Collagen Ⅰ were lower in overexpression of ALKBH5 group than those of the negative control group．CCK-8 assay and EdU suggested that the proliferation viability of CFs was reduced evidently in over- expression of ALKBH5 group，compared with the negative control group．@*Conclusion @#Overexpression of ALKBH5 can inhibit the proliferation of CFs，suggesting that ALKBH5 may be a key regulatory point in the development of myocardial fibrosis.

Efficacy comparison of three different arthroscopic techniques in treating Ellman Ⅱ partial articular surface tendon avulsions / 中华创伤杂志

Objective:To compare the clinical efficacy of arthroscopic debridement, transtendon repair and tear completion and repair in treating Ellman II partial articular surface tendon avulsions.Methods:A retrospective case control study was conducted on 45 patients with partial articular surface tendon avulsions admitted to East Division of Shanghai Sixth Hospital Affiliated to Shanghai Jiao Tong University from December 2017 through December 2019. There were 18 males and 27 females, with a mean age of (56.4±5.0)years (range, 38-66 years). The patients were assigned to arthroscopic debridement group ( n=15), arthroscopic transtendon repair group ( n=15), and arthroscopic tear completion and repair group ( n=15). Operation time and complications were recorded. American Shoulder and Elbow Surgeons (ASES) score and University of California, Los Angeles (UCLA) score were measured preoperatively, at postoperative 3 months and at the latest follow-up. Results:All the patients were followed up for 8-32 months [(18.2±6.3)months]. In arthroscopic debridement group, arthroscopic transtendon repair group and arthroscopic tear completion and repair group, the operation time was (43.7±3.1)minutes, (89.1±5.4)minutes, (62.2±3.6)minutes, respectively ( P<0.05). No complications such as nerve injury, infection and arthrofibrosis were observed after operation. In arthroscopic debridement group, arthroscopic transtendon repair group and arthroscopic tear completion and repair group, the ASES score was preoperative (48.7±2.9)points, (49.1±3.0)points, (48.3±3.6)points, respectively ( P>0.05); the UCLA score was preoperative (15.5±1.3)points, (15.4±1.8)points, (15.2±1.4)points, respectively ( P>0.05). In arthroscopic debridement group, arthroscopic transtendon repair group and arthroscopic tear completion and repair group, the ASES score at the latest follow-up was (90.1±1.6)points, (89.3±1.0)points, (89.4±1.0)points, respectively ( P>0.05); the UCLA score at the latest follow-up was (30.3±2.1)points, (29.0±2.3)points, (28.9±2.7)points, respectively( P>0.05). In arthroscopic debridement group, arthroscopic transtendon repair group and arthroscopic tear completion and repair group, the ASES score at postoperative 3 months was (84.4±2.3)points, (73.5±3.6)points, (73.6±3.2)points, respectively; the UCLA score at postoperative 3 months was (26.9±1.4)points, (21.4±1.0)points, (21.1±1.0)points, respectively. In comparison, no significant difference was found between arthroscopic transtendon group and arthroscopic tear completion and repair group ( P>0.05). However, ASES score and UCLA score in arthroscopic debridement group were significantly higher than those in arthroscopic transtendon group and arthroscopic tear completion and repair group ( P<0.05). Conclusions:Arthroscopic debridement, transtendon repair and tear completion and repair are effective techniques for treatment of Ellman II partial articular surface tendon avulsions. However, the operation time and early efficacy of arthroscopic debridement are better than other two techniques.

Drilling Combined with Adipose-derived Stem Cells and Bone Morphogenetic Protein-2 to Treat Femoral Head Epiphyseal Necrosis in Juvenile Rabbits / 华中科技大学学报（医学）（英德文版）

This study was designed to evaluate the effects of drilling through the growth plate and using adipose-derived stem cells (ADSCs) and bone morphogenetic protein-2 (BMP-2) to treat femoral head epiphyseal ischemic necrosis,which can be done in juvenile rabbits.Passage-four bromodeoxyuridine (BrdU)-labeled ADSCs were cultured,assayed with MTT to determine their viability and stained with alizarin red dye to determine their osteogenic ability.Two-month-old,healthy male rabbits (1.2 to 1.4 kg,n=45) underwent ischemic induction and were randomly divided into five groups (group A:animal model control;group B:drilling;group C:drilling & ADSCs;group D:drilling & BMP-2;and group E:drilling & ADSCs & BMP-2).Magnetic resonance imaging (MRI),X-ray imaging,hematoxylin and eosin staining and BrdU immunofluorescence detection were applied 4,6 and 10 weeks after treatment.Approximately 90％ of the ADSCs were labeled with BrdU and showed good viability and osteogenic ability.Similar results were observed in the rabbits in groups C and E at weeks 6 and 10.The animals of groups C and E demonstrated normal hip structure and improved femoral epiphyseal quotients and trabecular areas compared with those of the groups A and B (P＜0.01).Group D demonstrated improved femoral epiphyseal quotients and trabecular areas compared with those of groups A and B (P＜0.05).In summary,drilling through the growth plate combined with ADSC and BMP-2 treatments induced new bone formation and protected the femoral head epiphysis from collapsing in a juvenile rabbit model of femoral head epiphyseal ischemic necrosis.

Drilling Combined with Adipose-derived Stem Cells and Bone Morphogenetic Protein-2 to Treat Femoral Head Epiphyseal Necrosis in Juvenile Rabbits / 华中科技大学学报（医学）（英德文版）

This study was designed to evaluate the effects of drilling through the growth plate and using adipose-derived stem cells (ADSCs) and bone morphogenetic protein-2 (BMP-2) to treat femoral head epiphyseal ischemic necrosis,which can be done in juvenile rabbits.Passage-four bromodeoxyuridine (BrdU)-labeled ADSCs were cultured,assayed with MTT to determine their viability and stained with alizarin red dye to determine their osteogenic ability.Two-month-old,healthy male rabbits (1.2 to 1.4 kg,n=45) underwent ischemic induction and were randomly divided into five groups (group A:animal model control;group B:drilling;group C:drilling & ADSCs;group D:drilling & BMP-2;and group E:drilling & ADSCs & BMP-2).Magnetic resonance imaging (MRI),X-ray imaging,hematoxylin and eosin staining and BrdU immunofluorescence detection were applied 4,6 and 10 weeks after treatment.Approximately 90％ of the ADSCs were labeled with BrdU and showed good viability and osteogenic ability.Similar results were observed in the rabbits in groups C and E at weeks 6 and 10.The animals of groups C and E demonstrated normal hip structure and improved femoral epiphyseal quotients and trabecular areas compared with those of the groups A and B (P＜0.01).Group D demonstrated improved femoral epiphyseal quotients and trabecular areas compared with those of groups A and B (P＜0.05).In summary,drilling through the growth plate combined with ADSC and BMP-2 treatments induced new bone formation and protected the femoral head epiphysis from collapsing in a juvenile rabbit model of femoral head epiphyseal ischemic necrosis.

Analysis of needlestick and sharps injuries among medical staff in upper first-class hospital / 中华劳动卫生职业病杂志

<p><b>OBJECTIVE</b>To investigate the incidence of needlestick and sharps injuries among the medical staff in an upper first-class hospital and its risk factors and to provide a basis for the infection control department of hospital and health administration department to establish the policies for safe injection.</p><p><b>METHODS</b>A questionnaire survey was conducted in 800 healthcare professionals in an upper first-class hospital in Nanjing, China to investigate the incidence of needlestick and sharps injuries in the past year. A total of 641 persons completed the questionnaire, including 239 doctors, 348 nurses, and 54 other healthcare workers. The obtained data were subjected to chi-square test and logistic analysis.</p><p><b>RESULTS</b>Needlestick and sharps injuries occurred in 373 of 641 healthcare professionals, with an incidence rate of 58.19%. There were significant differences in the incidence of needlestick and sharps injuries among the medical staff of different types or in different departments (P < 0.01). The syringe needles, glass, scalp infusion needles, and suture needles were the major medical apparatus that caused needlestick and sharps injuries; the incidence rate of injury caused by suture needles in doctors (48.21%) was significantly higher than that in nurses (6.72%) (P < 0.01), and the incidence rates of injuries caused by syringe needles, scalp infusion needles, and glass in nurses (71.15%, 59.68%, and 49.04%) were significantly higher than those in doctors (46.43%, 6.25%, and 16.96%) (P < 0.01); compared with nurses, the doctors had higher incidence rates of injuries due to body fluid or tissue sample collection, wound flushing, suture, and cutting (P < 0.05); compared with doctors, the nurses had higher incidence rates of injuries due to injection, intravenous infusion, heparin cap sealing, intravenous infusion line connection, and venous or arterial blood collection (P < 0.05).</p><p><b>CONCLUSION</b>Needlestick and sharps injuries are common in medical staff. The results of the study on risk factors provide a basis for the infection control department of hospital and health administration department to establish the policies for safe injection.</p>

Investigation of work-related acute pesticide poisoning among farmers in Jiangsu Province and the risk factors / 中华劳动卫生职业病杂志

<p><b>OBJECTIVE</b>To determine the incidence of work-related acute pesticide poisoning among farmers in Jiangsu Province and to identify the risk factors.</p><p><b>METHODS</b>Multi-stage stratified sampling and cluster sampling methods were used to randomly select 1490 farmers from the south (Z village), centre (Y village) and north (X village) of Jiangsu Province. The database was established by EpiData 3.1. SPSS 17.0 and SAS 9.13 were used to do chi-square test, trend chi-square test, single factor and multi-factor logistic regression analysis.</p><p><b>RESULTS</b>Among 1490 farmers, 121 people had work-related acute pesticide poisoning in the past year. The incidence rate in Jiangsu Province was 8.1%. 118 farmers (97.5%) were mild pesticide poisoning.3 farmers (2.5%) were moderate pesticide poisoning. The incidence rate of Work-related acute pesticide poisoning in X village of North Jiangsu Province (13.5%, 68/505) is obviously higher than that in Y village of Central Jiangsu Province (4.8%, 19/399) and Z village of South Jiangsu Province (5.8%, 34/586), and the difference is statistically significant (χ(2) = 29.60, P < 0.01). The incidence rate of Work-related acute pesticide poisoning in female (10.4%, 58/557) is obviously higher than that in male (6.8%, 63/933), and the difference is statistically significant (χ(2) = 6.26, P < 0.05). There are lack of safety guidance, unsafety application methods (without alternate row spraying, without backward application, without down-wind application) and risk behaviors in the spraying (without read labels, preparing pesticides without gloves, wiping sweat with hand (s), equipment leakage, body pollution by pesticide, working when feeling sick, without bath after work). If farmers have risk behaviors above, the incidence rate of Work-related acute pesticide poisoning will be obviously higher than farmers who receive safety guidance and have no such dangerous behaviors, and all the differences are statistically significant (P < 0.05). The group of 24 ∼ 34 years old has the highest risk of work-related acute pesticide poisoning (OR = 7.15).</p><p><b>CONCLUSION</b>The incidence rate of work-related acute pesticide poisoning among farmers in Jiangsu Province was higher. Effective measures are needed to prevent and reduce the occurrence of work-related acute pesticide poisoning. We can teach the farmers who spray pesticides about safety guidance, safety application methods and avoiding risky behaviors.</p>

Autoregressive integrated moving average model in predicting road traffic injury in China / 中华流行病学杂志

This research aimed to explore the application of autoregressive integrated moving average (ARIMA) model of time series analysis in predicting road traffic injury (RTI) in China and to provide scientific evidence for the prevention and control of RTI.Database was created based on the data collected from monitoring sites in China from 1951 to 2011.The ARIMA model was made.Then it was used to predict RTI in 2012.The ARIMA model of the RTI cases was Yt=eYt-1+0.456▽Yt-1+et (et stands for random error).The residual error with 16 lags was white noise and the Ljung-Box test statistic for the model was no statistical significance.The model fitted the data well.True value of RTI cases in 2011 was within 95％ CI of predicted values obtained from present model.The model was used to predict value of RTI cases in 2012,and the predictor (95％CI) was 207 838 (107 579-401 536).The ARIMA model could fit the trend of RTI in China.

A case-control study on the risk factors of work-related acute pesticide poisoning among farmersfrom Jiangsu province / 中华流行病学杂志

Objective To explore the risk factors on cases regarding work-related acute pesticide poisoning among farmers of Jiangsu province.Methods A population-based,1 ∶ 2 matched case-control study was carried out,with 121 patients as case-group paired by 242 persons with same gender,district and age less then difference of 3 years,as controls.Cases were the ones who had suffered from work-related acute pesticide poisoning.A unified questionnaire was used.Data base was established by EpiData 3.1,and SPSS 16.0 was used for both data single factor and multi-conditional logistics regression analysis.Results Results from the single factor logistic regression analysis showed that the related risk factors were:lack of safety guidance,lack of readable labels befores praying pesticides,no regression during application,using hand to wipe sweat,using leaking knapsack,body contaminated during application and continuing to work when feeling ill after the contact of pesticides.Results from multi-conditional logistic regression analysis indicated that the lack of safety guidance (OR=2.25,95％ CI:1.35-3.74),no readable labels befores praying pesticides (OR=1.95,95％ CI:1.19-3.18),wiping the sweat by hand during application (OR=1.97,95％CI:1.20-3.24)and using leaking knapsack during application (OR=1.82,95％ CI:1.10-3.01) were risk factors for the occurrence of work-related acute pesticide poisoning.Conclusion The lack of safety guidance,no readable labels befores praying pesticides,wiping the sweat by hand or using leaking knapsack during application were correlated to the occurrence of work-related acute pesticide poisoning.

Characteristics of abnormal menstrual cycle and polycystic ovary syndrome in community and hospital populations / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Polycystic ovary syndrome (PCOS) is considered to be the most common endocrine disorder in women of reproductive age. The involvement of an abnormal menstrual cycle in the etiology of PCOS remains unclear. We aimed to analyze the characteristics of abnormal menstrual cycles and their association with PCOS in community and hospital patient populations.</p><p><b>METHODS</b>Women with PCOS identified from 2111 permanent female residents in the community of Beijing and 506 outpatients obtained from the reproductive clinic of Peking University Third Hospital were recruited for this study, comprising the PCOS community group and the PCOS hospital group, respectively. Each group was further divided into four subgroups according to the length of menstrual cycles: <21 days; 21-34 days; 35-60 days; and >60 days. Women in each group were interviewed using a questionnaire to assess factors including age, age of menarche, menstrual cycle history, related family history, and modified Ferriman-Gallwey (mF-G) score. All women received transvaginal ultrasound scan and had fasting blood samples taken for endocrine evaluation. A two-tailed P value of <0.05 was considered significant.</p><p><b>RESULTS</b>In the community population, the prevalence of abnormal menstrual cycle was 27.19% (574/2111). The prevalence of PCOS in the community was 6.11% (129/2111) according to Rotterdam criteria. In the community group, the most common menstrual cycle length was 35-60 days, whereas for the hospital group, it was >60 days. In both the community and hospital groups, the most common phenotype of PCOS was that of Oligo/amenorrhea+PCO+ hyperandrogenism (HA) (O+P+H) (P=0.000). With increasing cycle length of 35-60 days to >60 days, the percentages of Oligo/amenorrhea+PCO (O+P) and O+P+H were found to significantly decrease in the community group and significantly increase in the hospital group (P=0.000 for each). In the hospital group, as the menstrual cycle length increased from 35-60 days to >60 days, the rate of spontaneous abortion increased significantly (P=0.000), meanwhile the rate of poorly-secreted endometrium and abnormal endometrial hyperplasia increased significantly (P=0.000).</p><p><b>CONCLUSIONS</b>The prevalence of PCOS in the Beijing community of women was 6.11%. Oligo/amenorrhea was the most common type of abnormal menstrual cycle and may be an indicator for PCOS and endometrial lesions. Gynecologists should seek relevant medical information from women in the community to promptly diagnose PCOS and then follow up patients for potential development of subsequent complications.</p>

Color Doppler ultrasound in the diagnosis and treatment of arterial injury of extremities / 中国介入影像与治疗学

Objective To explore the clinical significance of the color Doppler ultrasound in diagnosis and treatment of artery injury of extremities. Methods Color Doppler ultrasound was used in 69 patients with limb trauma. Results Ultrasonography showed completely or partially vascular rupture in 13 cases (surgery confirmed 11 cases), artery injury and thrombosis in 18 cases (surgery confirmed 14 cases), highly suspicious vascular injuries in 19 cases (surgery confirmed 12 cases), vascular pressure, cramps or bone room fascia caused by high blood pressure obstacles in 19 cases (all confirmed afterclinical treatment). Conclusion Color Doppler ultrasound is the best approach for diagnosis and outcome evaluation of of artery injury of extremities.

Drug-Resistance of Staphylococcus in Nosocomial Infection / 中华医院感染学杂志

OBJECTIVE To study drug-resistance status in nosocomial infection with Staphylococcus in the local region,and take effective measures to control the nosocomial infection.METHODS Totally 129 strains of Staphylococcus were clinically isolated from various specimens from Jan 2006 to Dec 2008.The strain identification,detection of meticillin-resistant Staphylococcus(MRS) and K-B disk diffusion test were performed.The data were analyzed statistically.RESULTS Among all 129 strains consisting of 6 species of Staphylococcus,the isolating rate of S.aureus predominated with 39.5%.Among them,meticillin-resistant S.aureus(MRSA) accounted for 29.4%.Among all coagulase-negative Staphyloccocus(CNS),meticillin-resistant coagulase-negative Staphylococcus(MRCNS) accounted for 32.1%.Meticillin-susceptible S.aureus(MSSA) showed significantly lower drug-resistance than MRS.CONCLUSIONS Nosocomial infection status caused by MRS is serious in the grass-roots hospitals.Effective measures should be taken to control it.

Infection Status and Drug-susceptibility Analysis of Mycoplasma Strains Isolated from Genitourinary Tract / 中华医院感染学杂志

OBJECTIVE To investigate the infection status and drug resistance of Mycoplasma strains isolated from genitourinary tract. METHODS The IST kit produced by Bio-Merieux was used to culture the Mycoplasma strains and to do drug susceptibility test. RESULTS The Mycoplasma infection ratio was 41.0%,with Ureaplasma urealyticum(Uu) 71.9%,Mycoplasma hominis(Mh) 2.3%,and mixed infection 25.8%,respectively.The drug susceptibility test showed that drug resistance ratios were as follows: to erythromycin 58.7%,ciprofloxacin 50.0%,ofloxacin 41.3%,pristinamycin 3.3%,josamycin 5.4% and doxycycline 7.6%. CONCLUSIONS Mycoplasma infection in genitourinary tract is mainly due to Uu infection.We should rationally choose and use antibiotics according to the drug susceptibility,and josamycin is very effective.

Construction of a trans-splicing ribozyme for restoring EGFP truncation mutation / 生物工程学报

Special designed group I intron ribozymes can specifically splice objective RNA, repair the mutant gene in RNA level. The specificity of ribozyme is determined by nucleotides specific internal guide sequence (IGS) introduced to the enzyme. In this study, fragment sequence containing Tetrahymena thermophilia intron I of 26S rRNA gene was cloned and cis-splicing activity of this ribozyme was confirmed by in vitro transcription. For evaluating the trans-splicing activity of this ribozyme, a truncated mutant Green Fluorescence Protein (GFP) vector, XYQ5/XYQ10- pEGFP-C2, was constructed. This vector deleted the 3' end 564bp fragment of EGFP coding sequence, led to the lost the activity of emitting green fluorescence. Trans-splicing ribozyme plasmids ptrans-rib-CMV2 for remedy of the truncated mutant EGFP was constructed by PCR and molecular cloning techniques. This vector utilizing cloned 26S rRNA intron 1 as core enzyme; selecting T-G site at 194bp of EGFP coding sequence as splicing receptor, designed an IGS which is inversely complement to the 188-193nt of EGFP mRNA; the 195-890bp fragment of EGFP coding sequence was ligated to the 3'-end of ribozyme core. The fragment containing these components was inserted to a eukayotic expression vector pRC-CMV2. Using linearized XYQ5/XYQ10- pEGFP-C2 and ptrans-rib-CMV2 as templates, truncated EGFP mRNA and the constructed ribozyme vector were transcribed and mixed to evaluate the trans-splicing activity. Analysis of in vitro transcription products mix by RT-PCR verified the existence of wild type EGFP mRNA molecule. Co-transfection of XYQ5/XYQ10- pEGFP-C2 with ptrans-rib-CMV2 to Hela cells proved this ribozyme restored green fluorescence within cell, but the efficiency was low.

The construction of cDNA expression library from the tentacles of Sagartia rosea / 生物工程学报

A cDNA expression library of the tentacles of Sagartia rosea was constructed. The cDNA was cloned into eukaryotical expression plasmid pcDNA3. SMART protocol was used for cDNA library construction and bioinformatics analysis was carried out. 71 novel EST clones were obtained from 130 sequences in the library, of which there were 21 full-length clones, including cytolysin genes, flourescent protein, ubiquinol-cytochrome C reductase gene, elongation factor, ferritin gene riboflavin kinase gene, ribosomal protein. This provides a base for further investigating their biological activity and application.