Construction of recombinant plasmid vector for gene expression is a key step in making transgenic plants and important to study gene function and plant genetic engineering. A right choice of gene construction method can be cost-effective and achieve more diverse recombinant plasmids. In addition to the traditional methods in construction of plant gene expression vectors, such as Gateway technology, three DNA method and one step cloning, a few novel methods have been developed in recent years. These methods include oligonucleotide synthesis-based construction of small fragment gene expression vectors via competitive connection; construction of small RNA expression vector using pre-microRNA; recombination-fusion PCR method which inserts DNA fragments of multiple restriction sites into the target vector; and insertion of a DNA fragment into any region of a linear vector via In-Fusion Kit. Construction of complex vectors with many fragments uses sequence and ligation-independent cloning method, Gibson isothermal assembly or Golden Gate assembly. This paper summarizes our working experience in the area of recombinant vector construction and reports from others with an intention to disseminate ideas about currently widely used DNA recombination methods for plant transformation.
Cloning, Molecular ; DNA ; Gene Expression ; Genes, Plant ; Genetic Engineering ; methods ; Genetic Vectors ; Plants, Genetically Modified ; Plasmids ; Polymerase Chain Reaction

Objective To investigate the clinical efficacy of kidney-reinfocing and Du meridian-regulating acupuncture plus ginger moxibustion in treating ankylosing spondylitis. Methods Sixty patients with ankylosing spondylitis were randomly allocated to treatment and control groups, 30 cases each. The treatment group received kidney-reinfocing and Du meridian-regulating acupuncture plus ginger moxibustion and the control group, medication. After three months of treatment, the clinical therapeutic effects were compared between the two groups. Results The cure and marked efficacy rate and the total efficacy rate were 76.7%and 100.0%, respectively, in the treatment group and 53.3%and 70.0%, respectively, in the control group. There were statistically significant differences in both the rates between the two groups (P<0.05). Conclusion Kidney-reinfocing and Du meridian-regulating acupuncture plus ginger moxibustion is an effective way to treat ankylosing spondylitis.

AIM: To analyze the causes of blindness and low vision in patients with visual disability in Yangpu District of Shanghai from 2019 to 2022.METHODS:Cross-sectional study. A total of 1 604 patients who participated in the evaluation of visual disability in Shanghai Yangpu District Kongjiang Hospital, from April 2019 to December 2022 were selected for the study. The grade of visual disability and the main causes of blindness and low vision were determined by trained doctors.RESULTS:A total of 804 patients with visual disabilities were identified, with 87.31% aged 60 and above. The causes of visual disability were high myopic retinopathy(30.47%), age-related macular degeneration(23.26%), glaucoma(17.04%), and diabetic retinopathy(11.07%). Glaucoma(36.96%)is the leading cause of blindness.CONCLUSION: The majority of patients with visual disability are aged 60 years and above. More attention should be paid to the elderly population. Comprehensive prevention, treatment and rehabilitation measures should be applied in different diseases based on classification, so as to early reduce the occurrence of visual disability.

Objective To investigate the effects of neuromuscular electrical stimulation (NMES) combined with swallowing training for relieving dysphagia in stroke patients.Methods A total of 150 chronic stroke patients at least 3 months post-stroke who were afflicted with dysphagia were randomly and evenly divided into a conventional swallowing training group,an NMES group,and an NMES combined with conventional swallowing training group.Prior to and after treatment,surface electromyography (sEMG) signals from the swallowing muscles were detected,swallowing function was evaluated using the standardized swallowing assessment (SSA),their swallowing was studied videofluoroscopically (VFSS),and a swallowing-related quality of life questionnaire (SWAL-QOL) was administered.Results There were significant differences in the maximum amplitude of sEMG signals,and the scores on the SSA,VFSS and SWAL-QOL in both groups between prior to and after treatment.After 4 weeks of treatment,the maximum amplitude of the sEMG signals and all three assessment scores were significantly higher where NMES had been combined with conventional swallowing training group compared with the other two groups.Indeed,there was no significant difference between the group which had received conventional swallowing training and the NMES groups.The analysis of coefficients revealed correlation between sEMG maximum amplitude and the SSA,VFSS and SWALQOL results.Conclusions NMES combined with conventional swallowing training is helpful for improving swallowing function in chronic stroke patients with dysphagia.

Objective: To summarize the evolving concept and research progress on stability reconstruction in the surgical treatment of intertrochanteric fracture. Methods: Related literature and author's own experience concerning the surgical treatment of intertrochanteric fracture were reviewed and analyzed in terms of fracture pathoanatomy, stable and unstable pattern, adequate and in-adequate reduction, primary and secondary stability, postoperative stability evaluation, and early weight-bearing. Results: Intertrochanteric fracture occur at the translational area of cervico-trochanteric junction, which has a nature tendency to varus instability. Fracture reduction quality is the paramount factor and is evaluated by two views, the anteroposterior and lateral Garden alignment and cortex apposition between the head-neck fragment and the femoral shaft. Rather than the posteromedial lesser trochanteric frag ment, the cortical support concept (positive, neutral, negative) emphasizes the reduction of anteromedial cortex to a nonanatomic positive apposition or an "anatomic" neutral apposition in intraoperative fluoroscopy. Postoperative radiographic stability score provides a quantitative assessment for early weight-bearing standing and walking. However, some fractures may lose cortical contact and buttress (negative) during the process of postoperative telescoping and secondary stability. Further studies are needed to elucidate the risk factors such as tilting, swing or rotation of the head-neck fragment, and propose new preventive methods. Conclusion: Stability reconstruction of intertrochanteric fracture requires adequate fracture reduction with Garden alignment and anteromedial cortical support apposition, and reliable sustainment by internal fixation implants. Early weight-bearing standing and walking is safe in patients with perfect postoperative stability score.

Objective: To summarize the progress in the treatment of hyperextension tibial plateau fractures.

ObjectiveTo analyze the epidemiological characteristics of a family cluster of COVID-19 in a district of Shanghai， and to provide a scientific basis for prevention and control of cluster epidemic of COVID-19. MethodsField epidemiological survey was applied to study the cases and close contacts of a family cluster of COVID-19 in a district of Shanghai in 2021. Descriptive analysis of epidemiological survey data was conducted and real-time fluorescent quantification （RT-PCR） was used to detect new coronavirus nucleic acid for the collected specimens. ResultsCase A was the source of infection in the home aggregated epidemic. There were 2 second-generation cases infected in the whole incubation period. Case B and C were transmitted by case A through eating together. The PCR test results were negative both in the close contacts outside the family of primary contacts as well as in the close contacts to primary close contacts. ConclusionCOVID-19 is highly contagious and populations are generally susceptible. It is easy to cause family aggregated epidemic situation. Asymptomatic infection should be detected early and isolated. We should strengthen the tracking and management of close contacts of COVID-19 cases， and identify the second-generation cases as soon as possible.

Objective To ascertain the levels and distributions of individual dose of radiologic workers arising from external exposure in medical institutions in Yangpu District during 2018—2020, so as to provide basic data for the prevention of occupational radiation diseases. Methods Based on the individual dose monitoring specifications for occupational external exposure, the individual dose of radiologic workers was monitored by the thermoluminescence dosimetry systems, and the data were collected and analyzed. Results A total of 2082 people were monitored from 2018 to 2020, and the average annual effective dose per capital was 0.20 mSv·a⁻¹. The average annual effective dose per capital in 2020 was lower than that in 2018 and 2019, and the difference was statistically significant (P < 0.05). The average annual effective dose of 4 radiologic workers exceeded 5 mSv·a⁻¹ per capital. The average annual effective dose of diagnostic radiology was 0.20 mSv·a⁻¹, which was higher than that of dental radiology and other medical applications, and the difference was statistically significant (P < 0.05). The average annual effective dose per capital of nuclear medicine was 0.34 mSv·a⁻¹, which was higher than that of other medical categories, and the difference was statistically significant (P < 0.05). The average annual effective dose per capital in unrated hospitals was 0.14 mSv·a⁻¹, which was lower than that in tertiary hospitals and secondary hospitals, and the difference was statistically significant (P < 0.05). Conclusion The average annual effective dose per capital of radiologic workers in medical institutions in Yangpu District is generally kept at a low level. The radiation protection of radiologic workers in nuclear medicine, secondary and tertiary hospitals should be strengthened to reduce the occupational external exposure doses to the lowest level as possible.

OBJECTIVE：To investigate the effects of the third batc h of Shanghai drug quantity purchasing and national “4+7” drug quantity purchasing in a Shanghai hospital and its impact on the use of statins. METHODS ：Consumption amount （tablets） and sum of statins were collected from information system of this hospital before （Feb. 10th-Aug. 31th，2018）and after （Sept. 1th，2018-Mar. 19th，2019）the implementation of the third batch of Shanghai drug quantity purchasing ，after the implementation of national “4+7”drug quantity purchasing （Mar. 20th-Oct. 9th，2019）. The defined drug frequency （DDDs）and average defined daily cost （DDC） of above 3 periods were calculated according to drug defined daily dose （DDD） recommended by New Pharmacology and related drug instructions ，so as to analyzethe influence of the third batch of Shanghaidrug quantity purchasing and national “4+7”drug quantity purchasing on the use of statins in this hospital and predict the market share trend of each specification in this hospital. RESULTS & CONCLUSIONS ：Before the implementation of the third batch of Shanghaidrug quantity purchasing，there were 5 varieties and 8 specifications of statins in this hospital ；after the implementation of quantity purchasing ，it was adjusted to 5 varieties and 9 specifications. The amount of Simvastatin tablets was small in general ，and its consumption sum also continued to decline after the implementation of quantity purchasing. The amount of Rosuvastatin calcium tablets accounted for 49.49％ before the implementation of quantity purchasing ，ranking the first ；after the implementation of 3 batches of Shanghai drug quantity purchasing ， the amount proportion decreased to Δ 基金项目：上海市科学技术委员会科研计划项目（No.19ZR1450- 600） below 30％，and consumption sum proportion decreased from ＊本科 。研究方向 ：医药市场研究与分析 。电话 ：021- 37.82％ to 18.77％；after the implementation of national “4+ 54237308。E-mail：2123398711@qq.com 7”drug quantity purchasing ，the consumption sum proportion # 通信作者 ：副主任药师 ，博士。研究方向 ：临床药学 。电话： continued to decline to 17.38％ .The amount of Atorvastatin 021-65690520。E-mail：Hongjun.chen@tongji.edu.cn calcium tablets increased significantly after the implementation 中国药房 2021年第32卷第2期 China Pharmacy 2021Vol. 32 No. 2 ·247· of the thi rd batch of Shanghaidrug quantity purchasing ，from 41.55％ to 54.09％，and decreased slightly after the implementation of the national “4+7”drug quantity purchasing ，but the amount proportion was still as high as 53.06％，ranking the first ；before and after the implementation of quantity purchasing ，the proportions of its consumption sum were ranked the first ，which remained at about 50％，being 52.87％，61.18％ and 46.50％，respectively. Compared with the amount before the implementation of the third batch of Shanghaidrug quantity purchasing ，the amount of its generic drugs increased by 40.32％，but the consumption sum decreased by 78.18％. Any one type of Pravastatin sodium tablets and Pitavastatin calcium tablets was not selected ；the proportion of them was increased continuously ，but the overall proportion was relatively small. The price of statins decreased significantly after included in quantity purchasinglist. It is expected that the market share of simvastatin in thise hospital will continue to shrink ，and that of atorvastatin may continue to expand. Due to more adverse reactions ，rosuvastatin has limited room to expand ，and pravastatin accounts for a small proportion ，but there is still a demand for patients. Because pitavastatin is a new drug ，its application in this hospital needs more clinical data support.

OBJECTIVE: To compare the clinical effect of wheat grain moxibustion and warming acupuncture on chronic superficial gastritis of spleen-stomach deficiency and cold. METHODS: A total of 200 patients with chronic superficial gastritis of spleen-stomach deficiency and cold were randomly divided into a wheat grain moxibustion group (100 cases, 13 cases dropped off ) and a warming acupuncture group (100 cases, 16 cases dropped off ). The acupoints of Zhongwan (CV 12), Tianshu (ST 25), Qihai (CV 6), Liangqiu (ST 34) and Zusanli (ST 36) were selected in both groups. The patients in the wheat grain moxibustion group were treated with wheat grain moxibustion, and 5 cone were given on every acupoint each time. Moxibustion was performed after acupuncture in the warming acupuncture group, and 3 cone each acupoint. Both groups were treated every other day for 15 times. The scores of clinical symptoms and signs, scores of physical component summary (PCS) and mental component summary (MCS) of quality of life in the two groups were recorded before and after treatment and during follow-up 1 month after treatment, and the clinical effects of the two groups were evaluated after treatment and during follow-up. RESULTS: After treatment and during follow-up, the scores of clinical symptoms and signs of the two groups were lower than before treatment ( CONCLUSION The wheat grain moxibustion can effectively improve the clinical symptoms, signs and quality of life in patients with chronic superficial gastritis of spleen-stomach deficiency and cold, and its long-term effect is better than warming acupuncture.
Acupuncture Points ; Acupuncture Therapy ; Humans ; Moxibustion ; Quality of Life ; Spleen ; Stomach ; Triticum