Objective:To investigate the therapeutic effects of internal fixation via the posterolateral combined posteromedial approach in the treatment of posterior pilon fracture (Klammer type Ⅲ).Methods:A retrospective study was performed to analyze the 69 posterior pilon fractures (Klammer type Ⅲ) which had been treated by internal fixation with hollow screws or a buttress plate at Department of Orthopaedic Trauma, Yantai Shan Hospital from January 2015 to January 2020. There were 36 males and 33 females with an age of (45.3±10.0) years and duration from injury to surgery of (6.0±1.5) d. They were assigned into 2 groups according to different surgical approaches. The observation group (41 cases) was treated through the posterolateral combined posteromedial approach while the control group (28 cases) through the posterolateral approach alone. The therapeutic effects were evaluated by comparing the 2 groups in terms of incision length, intraoperative bleeding, operation time, fracture union time, fracture reduction (evaluated by the Burrwell-Charnley radiological score), the ankle-hindfoot score of American Orthopaedic Foot and Ankle Society (AOFAS) and complications.Results:There was no significant difference in the preoperative general data between the 2 groups, showing comparability ( P>0.05). All the 69 cases were followed up for (16.9±4.0) months, revealing primary healing of all the incisions and no vascular injury or wound infection. The incision length [(11.2±1.8) cm] and operation time [(76.0±6.6) min] in the observation group were significantly shorter than those in the control group [(12.4±1.9) cm and (79.7±6.8) min], the excellent and good rate of reduction in the observation group (97.6%, 40/41) was significantly higher than that in the control group (89.3%, 25/28), and the ankle-hindfoot scores of AOFAS at 1, 3 and 12 months after operation in the observation group [(78.4±5.6), (79.5±2.8) and (86.9±2.1)] were significantly higher than those in the control group [(75.2±5.5), (78.0±3.2) and (85.8±2.3)] (all P<0.05). There was no significant difference in the intraoperative bleeding between the 2 groups ( P>0.05). In the control group, 2 patients developed numbness in the dorsum of foot, which gradually disappeared after 3 months of treatment, but no other complications like persistent pain or flexor contracture within 1 year after operation. Conclusion:In the treatment of Klammer type Ⅲ posterior pilon fracture, the posterolateral combined posteromedial approach can result in satisfactory therapeutic effects, because the surgical approach can fully expose the fracture and facilitate better reduction.

Leiomyosarcoma of urinary bladder (LMS-UB) is a highly malignant mesenchymal tumor, accounting for less than 0.5% of all bladder malignancies, with a predominant clinical presentation of hematuria. Here we report a case of low-grade LMS-UB. A 44-year-old male patient was admitted to the hospital with urodynia for 2 weeks. The patient's pelvis CT showed a mass on the right part of the bladder. For this reason, he was initially diagnosed with bladder cancer. We performed a robot-assisted laparoscopic enucleation of the bladder tumor and low-grade LMS-UB was diagnosed with the histopathological examination. He underwent 5 cycles of adjuvant chemotherapy after surgery. At 19months postoperative follow-up, the patient had no symptoms, recurrence, or distant metastasis. There is no report on the treatment of LMS-UB with minimally invasive enucleation worldwide. This case provides a new comprehensive treatment method of enucleation combined with adjuvant chemotherapy for early low-grade LMS-UB to reduce complications and improve patients' quality of life after surgery.
Male ; Humans ; Adult ; Urinary Bladder/surgery* ; Leiomyosarcoma/secondary* ; Robotics ; Quality of Life ; Pelvis/pathology* ; Urinary Bladder Neoplasms/pathology* ; Laparoscopy/methods*

ObjectiveUncommon medicinal herbs are valuable medicinal resources， but their identification is a difficult problem in Chinese medicine due to their particularity and complexity. It is， therefore， urgent to establish a method for the identification of uncommon medicinal herbs. In this study， DNA signature sequence （DSS） tags were used to establish a specific polymerase chain reaction （PCR） identification method for Hibisci Cortex， the origin plant of Hibisci Cortex， and its adulterants. MethodThe candidate DSS tags were obtained from the chloroplast genome sequence analysis， and the DSS tags were verified by DNA sequencing. The specific identification primers for H. syriacus were designed based on the obtained reliable DSS tags. The PCR reaction conditions were optimized， and the tolerance and feasibility were investigated. ResultA DSS tag for identification of H. syriacus was obtained from the comparison of sequencing results of the amplified products with DSS， which revealed the distinguishing characteristics of Hibisci Cortex and its adulterants. A pair of specific primers for H. syriacus was designed according to the DSS tag. After PCR amplification and gel electrophoresis with the primers， a single bright band of about 270 bp was observed from H. syriacus， which did not appear in the four adulterants. ConclusionA DSS tag obtained in this study can be used to identify H. syriacus. The specific primers designed based on this DSS tag can accurately and simply identify the original plant of Hibisci Cortex and its adulterants， which provides a new method and idea for the molecular identification of genuine and counterfeit products of Hibisci Cortex.

As a naturally occurring steroid sapogenin, diosgenin acts as the precursor of hundreds of steroid medicines, and thereby has important medicinal value. Currently, industrial production of diosgenin relies primarily on chemical extraction from plant materials. Clearly, this strategy shows drawbacks of excessive reliance on plant materials and farmland as well as environment pollution. Due to development of metabolic engineering and synthetic biology, bio-production of diosgenin has garnered plenty of attention. Although the biosynthetic pathways of diosgenin have not been completely identified, in this review, we outline the identified biosynthetic pathways and key enzymes. In particular, we suggest heterologous biosynthesis of diosgenin in Saccharomyces cerevisiae. Overall, this review aims to provide valuable insights for future complete biosynthesis of diosgenin.
Biosynthetic Pathways/genetics* ; Diosgenin ; Metabolic Engineering

Objective:To disclose the genome characteristics and mutations of 2019 novel coronavirus (2019-nCoV) strains from the imported cases of Coronavirus Disease 2019 (COVID-19) in Gansu province, thereby to provide scientific reference for the prevention and control of COVID-19 in Gansu province. Methods:The respiratory tract specimens of imported COVID-19 cases from seven countries in Gansu province in 2020 were collected. The virus genome was sequenced by the second-generation sequencing technology, the whole genome sequences were compared and analyzed, and the MEGA software was used to construct a phylogenetic tree based on the neighbor-joining method.Results:A total of 46 2019-nCoV genome sequences with a length of 29 605~29 903 bp were obtained. Compared with the Wuhan reference strain (GenBank ID: NC_045512.2), it was found that the median (minimum to maximum) number of the nucleotide mutations of the 2019-nCoV genome sequence of the imported cases was 10 (7-24). A total of 134 nucleotide mutation sites were found in all 2019-nCoV genome sequences from 7 entry countries in Gansu province, distributed in 11 open reading frames (ORFs). The top three nucleotide mutations in different proteins: ORF1ab (78), S(20), N(12). Among the 134 nucleotide mutations, 82 caused amino acid mutations, and all of them were missense mutations. No insertions or deletions were seen. Types of deletion mutations, the top three amino acid mutations in different proteins: ORF1ab (46), S(11), N(10); the key sites of the receptor binding domain (RBD) of the S protein have not been mutated.Conclusions:No imported cases in Gansu province have been found to carry the reported mutations that can clearly lead to changes in the spread and pathogenicity of 2019-nCoV.

The antibiotics have obvious antibacterial and anti-inflammatory effects, but their toxic side effect, secondary infection and bacterial resistance have become a global problem. Traditional Chinese medicine(TCM) has always been the treasure of traditional culture and national characteristics in China since ancient times. It also has remarkable effect on inhibiting the growth of bacteria and killing pathogenic bacteria. The research on bacteriostatic experiment of TCM has gradually become a hot topic. Sensitivity experiments for such natural medicines have gradually become a research hotspot, but the complexity and particularity of natural medicines will vary with different methods. Therefore, different methods of drug sensitivity experiments should be matched with different natural drugs. By collecting and sorting out the relevant literature at home and abroad, this paper systematically summarizes the commonly used in vitro, in vivo and their combination bacteriostasis experimental methods of natural medicine activity, analyses the advantages and disadvantages of each method in the process of application, finds that different kinds of natural drugs have different applicable methods, and puts forward suggestions for the operation of each experimental method, in order to provide ideas for the selection of antibacterial susceptibility research experiments of natural medicines. It also provides a reliable reference method for solving the problem of antibiotic abuse and the development and utilization of natural medicines.

Objective To investigate the relationship between endoscopic diagnosis and pathological nature of gastric mucosal leukoplakia. Methods Analysis of gastric mucosal leukoplakia patients’ endoscopic features and pathological characteristics of last 3 years. Results Endoscopic diagnosis was made in 116 cases of gastric mucosal leukoplakia. Samples were gray white, round, round shaped or patchy in general observation. Diameter was 0.2 ~1.6 cm, foam cell aggregation appeared in the natural layer of mucous membrane in 88 cases (75.9 %), with varying degrees of inflammatory cells infiltration. The foam cells accumulation were observed in high rate among the gastric mucosal leukoplakia cases. Conclusion Pathological changes of gastric mucosal leukoplakia is gastric xanthoma with chronic inflammation of mucosa.

OBJECTIVETo investigate clinical and epidemiological characteristics of hospitalized severe acute respiratory illnesses (SARI) patients under 15 years old registered by sentinel hospitals at 10 cities and risk factors analysis of severe illness.

METHODSThe objects of this study were 2 937 SARI patients under 15 years old registered by sentinel surveillance in internal wards, pediatrics wards and intensive care units (ICU) of 10 sentinel hospitals in 10 cities during the period from December 2009 to June 2014. We also collected case report form (CRF) of them and their throat swabs for influenza testing. The inclusion criteria was hospitalized patients who were admitted by surveillance departments, registered by SARI surveillance system, under 15 years old, meeting SARI case definition and with complete CRF. Rank-sum test was used to compare the difference of age, the duration including from onset to admission, hospital stay and from onset to discharging/death between mild illness and severe illness. Chi-square test was used to compare the difference of demographic characteristics, influenza psoitive rate, vaccination rate of influenza, chronic medical conditions and clinical characteristics between mild illness and severe illness. Logistic regression was used to analysis risk factors associated with severe illness by two stratifications from SARI surveillance protocol (< 2 years old and ≥ 2 years old).

RESULTSAmong 2 937 SARI patients under 15 years old, 97.7% (2 872/2 937) was mild illnesses, and 2.3% (65/2 937) was severe illnesses. 78.8% (2 315/2 937) was under 5 years old. The median ages of severe illness and mild illness were 0.4 and 2.0 years old (U = -6.23, P < 0.001). The proportions of severe illness and mild illness with at least one chronic medical condition were 32.3% (21/65) and 8.4% (240/2 872) (χ² = 45.03, P < 0.001). The positive rate of influenza virus was 6.5% (190/2 937), which was 6.5% (186/2 858) for mild illness and 6.2% (4/65) for severe illness (χ² = 0.08, P = 0.961). The proportion of seasonal influenza vaccination was 1.5% (42/2 853), which was 1.5% (42/2 788) for mild illness and higher than that for severe illness (0) (χ² = 6.09, P = 0.048). For under 2 years old patients, age < 11 months and with at least one chronic medical condition were risk factors for severe SARI illness, and the risk for SARI patients who was 12-23 months and without medical condition was 14.71 (5.35-40.44) and 5.61 (2.96-10.63). For ≥ 2 years old patients, age, with at least one chronic medical condition and seasonal influenza vaccination history have no association with severe illness, OR (95% CI) was 0.92 (0.80-1.05), 0.67 (0.09-5.05) and 0.85 (0.31-2.35), respectively.

CONCLUSIONMost of SARI patients registered by 10 urban sentinel hospitals were patients under 5 years old. Age < 11 months and with at least chronic medical conditions were possible risk factors of severe illness of SARI patients.

Adolescent ; Child ; Child, Preschool ; China ; Chronic Disease ; Cities ; Hospitalization ; Hospitals ; Humans ; Infant ; Influenza, Human ; Orthomyxoviridae ; Respiratory Tract Diseases ; Risk Factors ; Sentinel Surveillance ; Vaccination

Objective To construct bispecific antibody BsAb1/17 against both IL-1β and IL-17A,express and purify the biologically active BsAbl/17 protein in prokaryotic system for further studies and applications. Methods VH1VL17-CL and VL1VH17-CH1 gene segments were constructed by overlap-PCR.Restriction enzyme sites Nco Ⅰ and BamH Ⅰ were designed at the 5'and 3' end primers respectively. The products of overlap-PCR were ligated to the Nco Ⅰ/BamH Ⅰ -prepared pET-27b vector. The recombinant plasmids pET-27b-VH1 VL17-CL(petA) and pET-27b-VL1 VH17-CH1 ( petB ) were transformed into E. coliRosetta separately. The expressing products were analyzed by SDS-PAGE and Western blot. Neutralization activity of the bispecific antibody for blocking the induction of IL-18 expression by IL-1β in human T cells was determined by real-time PCR. Neutralization activity of the bispecific antibody for blocking the induction of IL-6 expression by IL-17A in HeLa cells was determined by ELISA assay. Results The structure of the plasmids pET-27b-VH1 VL17-CL(petA) and pET-27b-VL1 VH17-CH1 (petB)was confirmed by DNA sequencing. After induction, the fusion proteins were expressed mainly as inclusion bodies. The purity of the both proteins exceeded 90%. SDS-PAGE analysis suggests the relative molecular mass of both products expressed by petA and petB were approximately 38× 103, which is in accordance with the theoretical value. The results of Western blot and ELISA test demonstrated that BsAb1/17 molecule had binding ability to both IL-1β and IL-17A. The BsAb1/17 could block IL-1β to stimulate human T cell to express IL-18 and block IL-17A to stimulate HeLa cell to express IL-6. Conclusion We successfully constructed a novel bispecific antibody BsAb1/17 against both IL-1 β and IL-17A, and expressed biologically active BsAb1/17 protein in prokaryotic system.

BACKGROUND:Recently,with the rapid development of material science and bioscience,the technology of dental implant has made great progress,especially the immediate implant technology.But the size and shape of implant are usually not fit for tooth extraction wound,so it is an important factor that leads to failure when implant and tooth extraction wound can not form close tangency.Guided bone regeneration or bone grafting materials are usually used to solve this problem.OBJECTIVE:To study the effects of nano-hydroxyapatite/collogen (nHAC) with recombinant human bone morphogenetic protein-2(rhBMP-2) and titanium (Ti) membrane on repairing peripheral bone defects of instant implant.DESIGN,TIME AND SETTING:A randomized,controlled animal study was performed at the Central Laboratory,the Fourth Military Medical University of Chinese PLA between January 2005 and January 2006.MATERIALS:Ti screw implants (diameter 2 mm,length 10 mm,and pitch 0.4 mm) without the part that went through gum were offered by Nonferrous Metal Academy in Baoji,China.The nonabsorbable Ti membranes (2 cm×2 cm) were offered by Zhongbang Biomaterial Limited Company in Xi'an,China.The nHAC materials were gifted by professor Cui Fu-zhai from Material Science and Engineering Department of Tsinghua University and fabricated into 0.5 mm×0.5 mm×0.5 mm small blocks.rhBMP-2 was offered by the Academy of Military Medical Sciences in Beijing,China.rhBMP-2 was dissolved with hydrochloric carbamidine and then nHAC was immersed in it.Vacuumization,freeze-drying,and Ekibon degermation were followed.Each gram of nHAC compounds required approximately 1 mg rhBMP-2.METHODS:Four healthy purebred male dogs were included in this study.According to the methods to repair bone defects rhBMP-2+Ti membrane,nHAC composited with rhBMP-2 was implanted,covering Ti membrane.Six defects were made on the mandible on each side.MAIN OUTCOME MEASURES:At 6 and 12 weeks after implantation,new bone formation and the correlation of new bone to Ti implant were investigated by radiography,bone density measurement and histological examinations.RESULTS:All the bone wounds were full of new bone at 12 weeks,the Ti implants were steady and the osseointegration was good.In the nHAC+rhBMP-2 and nHAC+rhBMP-2+Ti membrane groups,new bone formed and maturated earlier.In the nHAC +Ti membrane,blank control,and nHAC + rhBMP-2 +Ti membrane groups,new bone formed well below Ti membrane and the alveolar crest was plump.In the nHAC + rhBMP-2 +Ti membrane group,new bone formed earliest with the most quantity and the alveolar crest was the plumpest.By 12 weeks,the bone density had achieved the same level as the autologous cancellous bone transplantation,but the alveolar crest was plumper compared to the positive control group.In addition,bone mass was much more.Bone formed slowest in the blank control group and fastest in the positive control group with the crest of ridge being slightly absorbed.CONCLUSION:nHAC is a new good bone graft material in repairing peripheral bone defects of instant implant,especially compositing with rhBMP-2 and/or Ti membrane.A round method to repair the surrounding bone defect of instant implant should be selected according to the specific situation in the clinic.