Objective To develop and verify a loop-mediated isothermal amplification(LAMP)method for simultaneous detection of multiple mycoplasma contamination.Methods The conserved sequences of 16S rRNA genes of 13 mycoplasma species were aligned by the multiple sequence alignment tool of SnapGene software.Two internal primers FIP and BIP,two external primers F3 and B3 and loop primer LOOP(LF/LB)were designed according to the conserved sequences.The LAMP system was developed as follows:Bst DNA polymerase large segment(8 U/μL),10×ThermoPoly reaction buffer[20 mmol/L Tris-HCl,10 mmol/L KCl,10 mmol/L(H_4)_2SO_4,2 mmol/L MgSO_4 and 0.1%Triton X-100,pH 8.8],MgSO_4(100 mmol/L),dNTP Mix,FIP/BIP Primers,F3/B3 Primers,LOOP Primers,nuclease-free water and template DNA(13 plasmids carrying mycoplasma 16S rRNA).The products were identified by 1%agarose gel electrophoresis.The feasibility,specificity and repeatability of the method were verified,and the detection limit was determined.Results A total of 11 mycoplasma sequences were detected simultaneously by using two sets of LAMP primers screened.There was no nonspecific amplification in other nonspecific template detection,and the primer group had good specificity.The developed method showed good reproducibility and good sensitivity with the minimum detection limit of 30.1 copies/μL,and no false positive or false negative amplification.Conclusion LAMP technology can be used to detect a variety of mycoplasma contamination in cell culture,with good specificity and short time consumption,which can be used as one of the technical platforms for rapid detection of mycoplasma contamination in cell culture.

Objective To explore the diagnostic value of HR -HPV combined with TCT in the diagnosis of cervical cancer and precancerous lesion.Methods 1 198 cervical specimens were selected for the study.The results of HR -HPV detection and TCT detection were analyzed,with the pathological results as the gold standard,the diag-nostic value of HR -HPV and TCT in the diagnosis of cervical cancer and precancerous lesion were discussed. Results With increasing pathological grade,TCT positive rate increased gradually,there was a correlation between them(r =0.629,P <0.05).With increasing pathological grade,HR -HPV positive rate increased gradually,there was a correlation between them(r =0.684,P <0.05).The sensitivity of HR -HPV combined with TCT in the differ-ential diagnosis of cervical cancer,precancerous lesions was 100.00%,specificity was 79.64%,missed diagnosis rate was 0.00%,the misdiagnosis rate was 20.36%.Conclusion HR -HPV combined with TCT examination has high clinical value in the diagnosis of cervical cancer and precancerous lesions.

Ophthalmology belongs to microsurgical subject and anatomy is intricate. In clinical practice, students are required to skillfully use eye examination equipment. The foreign students from Medical School of Vienna University are eager to learn and know theoretical knowledge very well. Department of ophthalmology has developed specific and individualized teaching plans according to the characteristics and needs of foreign students. Through the configuration of high quality teachers, the practice teaching is ensured. Chinese and foreign students are encouraged to share their knowledge and form a study group. Opportunities for hands-on practice of ophthalmology are increased by using Wetlab condition. Problem-based heuristic teaching is used to stimulate students' learning initiative. Through these teaching measures, the students' quality of clinical practice has been enhanced, the teaching level of clinical ophthalmology has been improved and sustainable development has also been achieved.

Objective To evaluate the application value of ultrasound-guided Mammotome minimally invasive surgery system in non-palpable breast lesions.Methods A total of 325 lesions of 245 patients with breast lumps by ultrasound examination were under ultrasound-guided Mammotome minimally invasive surgery system.Results Accurate and complete resection of all of the 325 breast lumps was achieved,5 lesions with hematoma,the incidence rate was 1.54％ (5/325).The pathologic diagnoses included:fibro-adenoma (172 lesions),intraductal papilloma (8 lesions),fibroadenosis (87 lesions),fibro-cystic adenosis (56 lesions),invasive ductal carcinoma (2 lesions).Conclusion The application of ultrasound-guided Mammotome minimally invasive surgery system in non-palpable beast lesions has advantages of accurate positioning,minimally invasive,beautiful.

Objective To study the advantage of using axillary fascia swelling in cleaning the lymph nodes in modified radical mastectomy. Methods 866 patients with breast cancer collected from June 2005 to June 2011 were divided into the observation group (578 cases, axilla-ry fascia swelling method) and the control group (288 cases, standard operation method), and the postoperative complications of the two groups were compared. Results In the observation group, there were 8 cases (1. 4%) of upper limb lymphatic edema, 13 cases (2. 2%) of chest wall and upper limb paresthesia, and 8 cases (1. 4%) of lymphatic leakage. The complications of the observation group were significantly reduced compared with the control group (P<0. 05), and there was no increase in the local recurrence rate (P<0. 05). Conclusion Axillary fascia swell-ing applied in cleaning lymph nodes could obviously reduce the postoperative complications after modified radical mastectomy.

Objective Membranous nephropathy ( MN) is rarely complicated by crescents.This study was to observe the clinicopathologic characteristics of MN with crescents. Methods This retrospective study included 53 cases of biopsy-proven idiopathic MN with crescents in the absence of immunologic and clinical etiologic factors and another 100 MN patients without histological crescents as controls.The clinicopathologic features, treatment response, and out-comes were analyzed and compared between the two groups of pa-tients. Results Significantly higher percentages of hypertension and decreased eGFR were observed in the MN +crescents group than in the control (47.2%vs 19.0%, P<0.05;28.3%vs 40.%, P<0.05).Circulating autoantibodies against the M-type phospholipase A2 receptor (PLA2R) were found in 66.7%(30/45) of the patients.The glomeruli exhibited a median of 4.6%(1.8%-35.3%) involvement of crescents.Compared with the controls, the MN +crescents group showed remarkably higher rates of segmental glomer-ulosclerosis lesions (16.0%vs 49.1%, P<0.05), capillary loops necrosis (0.0%vs 11.3%, P<0.05), interstitial fibrosis/tubu-lar atrophy (IFTA) (54.0%vs 86.8%, P<0.05) and afferent arterial lesions (65.0%vs 92.5%, P<0.05).No significant differ-ences were found in the outcomes of the two groups of patients. Conclusion MN with crescents is rare, and secondary MN and cres-centic glomerulonephritis should be considered.Crescentic MN usually presents with hypertension and renal dysfunction clinically and high rates of severe segmental and global glomerulosclerosis, capillary loops necrosis, and IFTA histologically.The condition has a fa-vorable short-term prognosis.

ObjectiveTo define the maximum tolerated dose (MTD) of weekly cisplatin in concurrent chemoradiotherapy for Chinese cervical carcinoma.MethodsCervical carcinoma of stage ⅠB2- ⅣA were eligible for the study.PhaseⅠstudy was dose-escalation trial with 15 patients.All patients received whole pelvic radiotherapy with three dimentional conformal radiotherapy technique. Concurrent cisplatin started from the dose of 20 mg/m2 to 25 mg/m2,30 mg/m2,35 mg/m2,40 mg/m2 for the weekly schedule ( ≥3 patients per dose group) and the doses were steadily escalated to 40 mg/.m2.If the dose was increased to 40 mg/m2 without dose-limiting toxicity ( DLT),40 mg/m2 would be the maximum tolerated dose (MTD).According to the MTD dose from Phase Ⅰ study,we conducted phase Ⅱ clinical trial with 36 patients.ResultsIn Phase Ⅰ study,cisplatin dose was escalated to 40 mg/m2 and DLT had not been reached.Thirty-six patients in Phase Ⅱ study included 9 inpatients and 27 outpatients.All 9 inpatients completed 6 cycles of chemotherapy. In 27 outpatients,18 patients (66％) completed 6 cycles of chemotherapy,19 patients (70％) completed 5 cycles and 25 patients (92％) completed 4 cycles of chemotherapy.All patients completed radiotherapy.Major adverse effects were grade 1 and 2 gastrointestinal toxicities and neutropenia.ConclusionsWeekly 40 mg/m2 cisplatin concurrent with radiotherapy is well tolerated when given to Chinese patients with cervical carcinoma. For outpatients with poor performance status,the cisplatin dose needs to be reduced.

Objective To investigate the current staffing of nurses and need of hospital nursing human resources in China.Methods Data were collected from 181 secondary and tertiary hospitals and 9774 nurses in mainland China by questionnaires.Results The average doctor-nurse ratio was 1 ∶ 1.39 in tertiary hospitals and 1 ∶ 1.31 in secondary hospitals.The doctor-nurse ratios in 164 hospitals (97.6％) and the proportion of nurses in health care staff in 105 hospitals (61.8％) had not reached the standard set by the Ministry of Health of China.62.3％ nurses held secondary diploma for their initial nursing education.The constituent ratio of nurses held secondary diploma decreased,while the ratio of nurses held advanced diploma and bachelor degree increased in the last 5 years from 2003 to 2007.Sixty percent of newly employed nurses were contract nurses in 2003.The proportion increased to 78％ in 2007,and in some regions it accounted for more than 90％ of new nurses.The needs and constituent ratio of nurses with.secondary diploma and advanced diploma would decrease while nurses with bachelor degree and master degree would increase in the next 5 years (from 2009 to 2013).Conclusions The nursing shortage is still severe in China,and nursing staff mainly held secondary diploma for their initial nursing education.Most of new nurses were employed as conwact nurses.The need of hospitals for nurses has increased,especially for nurses with higher educational level such as bachelor degree and master degree,while the need for secondary diploma hold ers have decreased.The need for advanced diploma holders has increased in the last 5 years and would decrease in the next 5 years,but they would still be the majority of employed nurses.The authors suggested that the staffing of nursing manpower,the work environments and career development of contract nurses should be improved,and the initial nursing education should be upgraded to meet the needs of hospitals.

The aims of this study were to evaluate the expression of enhanced green fluorescent protein (EGFP) driven by 6 different promoters, including cytomegalovirus IE enhancer and chicken beta-actin promoter (CAG), cytomegalovirus promoter (CMV), neuron-specific enolase promoter (NSE), myosin 7A promoter (Myo), elongation factor 1alpha promoter (EF-1alpha), and Rous sarcoma virus promoter (RSV), and assess the dose response of CAG promoter to transgene expression in the cochlea. Serotype 1 adeno-associated virus (AAV1) vectors with various constructs were transduced into the cochleae, and the level of EGFP expression was examined. We found the highest EGFP expression in the inner hair cells and other cochlear cells when CAG promoter was used. The CMV and NSE promoter drove the higher EGFP expression, but only a marginal activity was observed in EF-1alpha promoter driven constructs. RSV promoter failed to driven the EGFP expression. Myo promoter driven EGFP was exclusively expressed in the inner hair cells of the cochlea. When driven by CAG promoter, reporter gene expression was detected in inner hair cells at a dose as low as 3 x 10(7) genome copies, and continued to increase in a dose- dependent manner. Our data showed that individual promoter has different ability to drive reporter gene expression in the cochlear cells. Our results might provide important information with regard to the role of promoters in regulating transgene expression and for the proper design of vectors for gene expression and gene therapy.
Animals ; Cochlea/cytology/*metabolism ; Dependovirus/*genetics ; Dose-Response Relationship, Drug ; Female ; Genetic Vectors/*genetics ; Green Fluorescent Proteins/metabolism ; Humans ; Mice ; Mice, Inbred C57BL ; Promoter Regions, Genetic/*genetics ; *Transgenes

Objective: To conduct an epidemiological investigation on a cluster of overseas imported cases of coronavirus disease 2019(COVID-19)in Qingtian County, Lishui,so as to provide reference for the prevention and control of COVID-19 imported from abroad. Methods: According to the COVID-19 Prevention and Control Program (FiFth Edition) issued by National Health Commission of China,a field investigation was employed and the close contacts of the case were tracked down ;the real-time reverse transcription polymerase chain reaction (RT-PCR) assay was performed to detect the nucleic acid of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) from the sputum specimens or throat swabs of cases;the epidemiological characteristics,source of infection,route of transmission and disposal of the cluster were analyzed. Results: From March 1 to March 6,2020,eight confirmed cases and one asymptomatic case of COVID-19 were reported. Their median age was 33 years old. The nine cases all had no fever,no decrease in leukocyte and lymphocyte levels,and the clinical symptoms of seven cases were not obvious. The asymptomatic case had been tested negative for SARS-CoV-2 for six times,but had been weakly positive for IgM antibody and strongly positive for IgG antibody. Nine cases were all the employees of the same restaurant in Begamo,Italy. They lived in three independent villas and usually had lunch and dinner in the restaurant where they worked. Begamo had COVID-19 epidemic,but the staff of the restaurant did not take any protective measures such as wearing masks and environmental disinfection. Eight cases reported to have cold symptoms in Italy during mid February. Through a closed-loop management of“all people,all sites,all chains ”,such as treatment of current cases,screening and isolation of close contacts and health education,totally 372 close contacts were traced back,yet no fever or respiratory symptoms have been found. Conclusions It was a cluster of COVID-19 cases imported from abroad. The clinical symptoms of the cases were not obvious. Qingtian County adopted the closed-loop management in time,and effectively controlled the spread of the epidemic. No second-generation cases have been found.