Objective:To investigate the prevalence of enterovirus 71 (EV71) and the genetic characteristics of VP1 region in common hand, foot and mouth disease (HFMD) cases in Shenzhen from 2016 to 2022.Methods:Throat swabs from mild HFMD in Shenzhen sentinel hospitals were collected from 2016 to 2022. A total of 38 EV71-positive samples were screened from these throat swabs and were sequenced. Then, the VP1 sequence of these EV71-positive samples were analyzed for their phylogenetic evolution by bioimformatics software DNAStar and MEGA 6.Results:From 2016 to 2022, the number of EV71 infections among HFMD patients in Shenzhen sentinel hospitals decreased from 136 in 2016 to 0 in 2022. The mumber of EV71 infections in 2018 and 2019 decreased by 96.3%(257/267) compared to that in 2016 and 2017. From 2020 to 2022, the number of EV71 infections decreased to 0. During this period, the EV71 vaccination rate among HFMD patients increased from 6.4% to 39.6%; Evolutionary analysis showed that the nucleotide homology and amino acid homology between 38 EV71 sample strains in Shenzhen from 2016 to 2022 were 91.8%-99.9% and 98.3%-100.0%, all belonging to the C4a subgenotype; Among them, 26 strains wene local epidemic strains, and 11 strains were imported from other provinces, with a close genetic relationship with epidemic strains in Hainan, Yunnan, Sichuan, Tianjin, Henan, Jilin, and other places. One strain from 2017 had the closest genetic relationship with the US epidemic strain OP207969-USA-2017. Further comparing the EV71 epidemic strains in Shenzhen from 2016 to 2022 and EV71 severe strains, it was found that the EV71 strains in Shenzhen carried four amino acid mutation sites related to severe condition, named R22H, K43R, I249V and T289A.Conclusions:The EV71 epidemic strains in Shenzhen from 2016 to 2022 all belong to the C4a subgenotype, and the number of EV71 infection shows a downward trend with the increase of vaccine coverage rate. At the same time, the distribution of EV71 virus strains in Shenzhen shows a significant decrease in local strains and a predominance of imported strains. There are a total of four amino acid mutation sites associated with severe cases in the EV71 sample strains in Shenzhen from 2016 to 2022. Among them, 22R and 289T are located at the N and C ends of VP1, which are related to EV71 adsorption and targeting cells. The 43R site is associated with binding ability to Annexin2 protein, which enhances cell binding ability.

  Objective  To evaluate the clinical outcomes and complications of posterior spinal fusion surgery in the treatment of neurofibromatosis type 1(NF1)thoracolumbar kyphoscoliosis, and to explore the mode of perioperative care for nurses provided to the patients.  Methods  We used the retrospective analysis on the 134 patients with NF1 thoracolumbar kyphoscoliosis admitted to our department from March 2012 to April 2022 and analyzed the clinical outcomes and perioperative complications by using the related statistics. We evaluated the Perioperative care by the nurses in the treatment of NF1 to explore the mode of nursing to the patients with the NF1, by using specific observation points and evaluation indicators.  Results  NF1 kyphoscoliosis patients had poor preoperative nutritional status and lung function. NF1 kyphoscoliosis underwent longer operation time, lost more blood in operation, had higher osteotomy grade and more postoperative complications. All the patients successfully completed the operation in our group. The correction rate of scoliosis was (52.8±22.7)%, and the correction rate of kyphosis was (57.3±34.6)%. 25 patients had complications but no such serious complications as nerve damage.  Conclusions  The practice of the perioperative nursing to NF1 type scoliosis patients facilitates the shortening of the recovery period, the prevention or timely detection of complications, and improvement of the therapeutic effect.

Objective:To observe the effect of hypoxia on the dedifferentiation process of chondrocytes in vitro and explore the role of collagen prolyl 4-hydroxylase (C-P4Hs).Methods:Chondrocytes were treated with COCl 2 for different concentrations, and selecting the optimal COCl 2 concentration for hypoxia inductionwas100 μmol/L, the mouse costal chondrocytes were divided into the normal oxygen group and the hypoxia group, and the indexes of the 3rd generation 0.5-72 h and the 1st, 3rd, 5th and 7th generation at 72 h were detected respectively. The proliferation rate was determined by CCK8 method and cell count, and the dynamic changes of HIF-1α, P4Hα1, P4Hα2 and Col II in each group were detected by RT-qPCR, IF and Western blot. Results:Costal chondrocytes were cultured under different concentration of COCl 2 for 48 h. When COCl 2>150 μmol/L, the proliferation rate ( P<0.05) was significantly decreased. Normal oxygen and hypoxia induced rib chondrocytes for 0-72 h, and RT-qPCR showed significant increases in P4Hα2 and Col II mRNA in hypoxia group ( P<0.05). IF showed that HIF-1α and P4Hα2 accumulated in the nucleus under hypoxia, and P4Hα2 gradually entered the cytoplasm from the nucleus. Westernblot analysis showed that HIF-1α and P4Hα2 protein expressions were significantly increased in hypoxia group ( P<0.05). The expression of Col II protein in hypoxia group ( P<0.05) increased at the induction stage. CCK8 and cell count results showed that the proliferation rate and cell number of each generation in the hypoxic group were significantly increased ( P<0.05), and there was still potential for proliferation when the cells were transferred to the 6-7 generation. RT-qPCR showed that hypoxia group each generation cells P4Hα2, Col II mRNΑ were significantly increased ( P<0.05). Westernblot results showed that HIF-1α, P4Hα2 and Col II protein expressions were increased in each generation of hypoxia group ( P<0.05). ConcIusion:Increased expression of P4Hα2 through hypoxia induced HIF-1α can accelerate post-translational modification of Col II in chondrocytes and increase synthesis and accumulation of Col II. P4Hα2 may be responsible for increased proliferation rate and delayed dedifferentiation of chondrocytes cultured in vitro under hypoxia condition.

Objective To compare the effect of closed reduction internal fixation and open reduction and internal fixation on the recovery of double ankle joint function and the degree of pain in patients with comminuted double malleolus fracture after operation.Methods From June 2015 to March 2017,78 patients with comminuted malleolus fracture in the Department of Orthopedics from Changzhi People's Hospital Affiliated to Shanxi Medical University were randomly divided into two groups according to the digital table ,with 39 cases in each group.The control group was treated with open reduction and internal fixation ,and the study group was treated with closed reduction and internal fixation.The amount of bleeding ,operation time,time of hospitalization were compared between the two groups.Six months after operation,the ankle function and pain score were used to evaluate the clinical efficacy.Results All the operations of 78 patients were successfully performed.The operative blood loss,operative time and hospitalization time in the study group were significantly lower than those in the control group (t＝4.65,14.63,7.83,all P＜0.05).Six months after operation,the excellent and good rate of the double ankle function in the study group (97.44%) was significantly higher than that in the control group (74.36%)(χ2＝6.77,P＝0.01).The pain scores of the two groups were significantly lower than those before treatment (t＝24.84,33.81,all P＜0.01).Six months after treatment,the pain score in the study group[(1.74 ±0.55)points]was significantly lower than that of the control group [(2.78 ± 0.80)points] (t＝6.69,P＜0.01).Conclusion Closed reduction and internal fixation for the patients with commi-nuted malleolus fracture after the operation of the joint without reduction ,are beneficial to promote the recovery of the double ankle joint function and reduce the pain degree ,and have the advantages of small wound ,less bleeding in the operation,short operation time and short hospitalization time.The clinical application should be popularized.

Through introducing mutations into ribosomes by obtaining spontaneous drug resistance of microorganisms, ribosome engineering technology is an effective approach to develop mutant strains that overproduce secondary metabolites. In this study, ribosome engineering was used to improve the yield of butenyl-spinosyns produced by Saccharopolyspora pogona by screening streptomycin resistant mutants. The yields of butenyl-spinosyns were then analyzed and compared with the parent strain. Among the mutants, S13 displayed the greatest increase in the yield of butenyl-spinosyns, which was 1.79 fold higher than that in the parent strain. Further analysis of the metabolite profile of S13 by mass spectrometry lead to the discovery of Spinosyn α1, which was absent from the parent strain. DNA sequencing showed that there existed two point mutations in the conserved regions of rpsL gene which encodes ribosomal protein S12 in S13. The mutations occurred a C to A and a C to T transversion mutations occurred at nucleotide pair 314 and 320 respectively, which resulted in the mutations of Proline (105) to Gultamine and Alanine (107) to Valine. It also demonstrated that S13 exhibited genetic stability even after five passages.
Genetic Engineering ; Macrolides ; metabolism ; Point Mutation ; Ribosomal Proteins ; genetics ; Ribosomes ; metabolism ; Saccharopolyspora ; metabolism

BACKGROUND:With the development of sports medicine and research of radiologic imaging techniques, Blumensaat line (the radio-opaque line at the roof of the intercondylar notch) has been paid increasing attention. Blumensaat line is considered as measurement indexes of knee diseases. Taking advantage of the Blumensaat line, many surgeons and radiography physicians are trying to diagnose some knee diseases. OBJECTIVE:According to the knowledge about Blumensaat line in auxiliary diagnosis of knee disease, we hoped that it wil have a wide application in clinic. METHODS:A computer-based online search of CBM, CNKI, Wangfang Database and PubMed between 2000 and 2015 was performed for articles addressing Blumensaat line. We summarized its application as different diagnostic indicators. The key words were patel a alta, Blumensaat line, anterior cruciate ligament (ACL) injuries and ACL reconstruction. Thirty-nine studies were accorded with the inclusion criteria. RESULTS AND CONCLUSION:Blumensaat line represents the tangential y hit part of the roof in the intercondylar fossa. The line can be used for diagnosing ACL injuries and directing ACL restruction. (1) Blumensaat line and patel a alta:Patel a heights can be measured with the use of Blumensaat method, modified Blumensaat method and modified Blumensaat ratio. Modified Blumensaat ratio was found by Japanese researchers in 2014 and it is efficient. (2) Blumensaat line and ACL injuries:Blumensaat angle is formed by Blumensaat line and ACL. If this angle is negative or it is greater than 15°, we can draw a conclusion that the ACL was hurt. (3) Harner’s method can be used for choosing an accurate isometric point and a perfect bone tunnel’s angel in ACL reconstruction.

BACKGROUND:Proximal femoral nail anti-rotation has good biomechanical basis, and has obvious advantages for intertrochanteric fracture in aged patients, but there are some problems in the clinic, because of improper handling of material matching and operation details, which can impact therapeutic effects and functional recovery. OBJECTIVE:To analyze the efficacy and issues of proximal femoral nail anti-rotation in the treatment of intertrochanteric fracture in patients at more than 60 years old. METHODS:From July 2011 to July 2012, proximal femoral nail anti-rotation was used to treat 56 cases of intertrochanteric fractures. Clinical data bank was established to analyze intraopeative problems and postoperative complications. At 1, 3, 6, 9 and 12 months postoperatively, outpatient and telephone folow-up were carried out to evaluate therapeutic effects and functional recovery of hip joint. RESULTS AND CONCLUSION:Four patients died within 1 year. Seven patients lost within a year for other reasons. The remaining 45 patients were folowed with the time from 12 to 24 months, with an average time of 18.2 months. Harris score was (85.00±6.75) points. There were excelent in 26 cases, good in 15 cases, average in 3 cases and poor in 1 case, with an excelent and good rate of 91%. 18 cases were not satisfied with the position of fracture fragments. In 9 cases, proximal femur was not match with the proximal femoral nail anti-rotation. Seven cases were not satisfied because of the location and length of the spiral blade. Seven cases affected lateral cortex fracture. One case experienced postoperative pulmonary embolism. One case suffered from cardiovascular and cerebrovascular diseases. Nine cases suffered from local sweling. 13 cases experienced hip pain. Five cases affected the healing of fracture extended. Results showed that proximal femoral nail anti-rotation for intertrochanteric fracture in aged patients obtained good outcomes, but we should improve the separation of fracture fragments and reduce intraoperative and postoperative complications.

In order to clarify the chemical constituents of Si-Wu Decoction rapidly and holistically, we analyzed the ethanol extract of Si-Wu Decoction by UPLC/Q-TOF-MSE and UNIFI which based on traditional Chinese medicine database, the probable structures of 113 compounds were identified. The results show that this method can rapidly and effectively characterize the chemical compounds of Si-Wu Decoction and provide a new solution for identification of components from complex TCM extract.

BACKGROUND:The main component of cartilage, type Ⅱ col agen gene expression in chondrocyte is positively correlated with SOX9 concentration in a dose-dependent manner.
OBJECTIVE:To observe the variation of SOX9 and type Ⅱ col agen mRNA content at different periods in the differentiation process (osteogenic, chondrogenic, adipogenic induction) of mesenchymal stem cel s, and to explore the correlation of SOX9 expression and type Ⅱ col agen.
METHODS:Bone marrow mesenchymal stem cel s were isolated from 4-week-old Kunming mice, and cultured in vitro to passage 3. The cel phenotype was identified with flow cytometry. Cel s were divided into three groups and subjected to three kinds of induction conditions favorable for adipogenic, chondrogenic and osteogenic differentiation, and each group was observed at three time points. In addition, the non-induced cel s were used as a control group. The total RNA of cel s was extracted at 3, 7, 14 days after induction, and SOX9 and type Ⅱ col agen mRNA was quantified with reverse transcription-polymerase chain reaction. The induced cel s were stained by immunofluorescence to observe the differentiation and perform statistical analysis.
RESULTS AND CONCLUSION:Passage 3 bone marrow mesenchymal stem cel s grew wel , and cel phenotype was confirmed as stem cel s by flow cytometry. The staining results showed that, the cel s differentiated into chondrocytes, adipocytes and osteoblasts. The SOX9 mRNA levels in the induced cel s were the highest in chondrogenic differentiation group, then in osteogenic differentiation group, and the lowest in adipogenic differentiation group. Type Ⅱ col agen mRNA levels in the induced cel s were the highest in chondrogenic differentiation group, then in adipogenic differentiation group, and the lowest in osteogenic differentiation group. SOX9 expression in chondrogenic differentiation group increased at 3 and 7 days, and then decreased at 14 days. While type Ⅱ col agen expression increased at 3, 7, 14 days. SOX9 mRNA levels increased as the osteogenic differentiation, while type Ⅱ col agen expression gradual y decreased. There was no significant difference in the SOX9 mRNA expression between adipogenic differentiation group and control group (P>0.05), while type Ⅱ col agen expression was not regularly changed. Experimental findings suggest that, critical effect of SOX9 in chondrogenic differentiation is better than that in osteogenic and adipogenic differentiation. SOX9 is associated with type Ⅱcol agen, which may alter along with the SOX9 in the early chondrogenic differentiation;SOX9 may play a fine-tuning role in the process of chondrogenic and osteogenic differentiation.

Due to the insufficient supply of embryonated chicken eggs,the preparation of large quantities of inactivated influenza vaccines will require an alternative virus culture system after the emergence or reemergence of a pandemic influenza virus.The Vero cell is one of the ideal options since it was used for producing many kinds of human vaccines.However,most of the influenza viruses can not grow well in Vero cells.To develop a new influenza vaccine with Vero cells as a substrate,the virus needs to adapt to this cell substrate to maintain high growth characteristics.By serial passages in Vero cells,the B/Yunnan/2/2005va(B)strain was successfully adapted to Vero cells,with the hemagglutination titer(HAT)of the virus reaching 1:512.The high growth characteristic of this strain is stable up to 21 passages.The strain was identified by hemagglutination inhibition (HAI)test and sequencing respectively;the HA;gene sequence of the virus was cloned and analyzed.The screening and establishment of high growth B virus provides an important tool for influenza vaccine production in Vero cells.