Objective To isolate the Japanese encephalitis virus carried by Culex tritaeniorhynchus in Dongchuan District of Yunnan Province and analyze its molecular characteristics, so as to provide insights into the prevention and control of Japanese encephalitis in Yunnan Province. Methods Mosquito specimens were collected using mosquito-trapping lamps from pig farms in Batang Village and Xiaoxin Village, Dongchuan District, Kunming City, Yunnan Province in July 2016, and the mosquito species was identified according to the mosquito morphology. Then, 60 to 100 mosquitoes of each species served as a group and were ground. Baby hamster kidney-21 (BHK-21) cells and Aedes albopictus clone C6/36 cells were used for virus isolation, and positive isolates were identified using flavivirus primers. The positive isolates were amplified using reverse transcription polymerase chain reaction (RT-PCR) assay with 15 pairs of specific primers covering the full length of the genotype I Japanese encephalitis virus, and DNA sequence assembly was performed using the software SeqMan in the DNASTAR package. The obtained sequences were aligned with the complete sequences of 38 Japanese encephalitis virus downloaded from the GenBank with the software MegAlign, and the nucleotide and amino acid homology analyses of the obtained sequences were performed. The difference in amino acid sites was analyzed with the software GeneDoc, and phylogenetic trees were created based on the sequences of the coding region and E protein of the isolated Japanese encephalitis virus with the software Mega X. In addition, the secondary and tertiary structures of the E protein of the Japanese encephalitis virus were predicted using the online tool SOPMA and the software Swiss-Model. Results A total of 5 820 mosquitoes were collected and 3 843 Cx. tritaeniorhynchus (66.03%) were identified according to the mosquito morphology. A positive virus isolate, termed YNDC55-33, was isolated from Cx. tritaeniorhynchoides following batches of virus isolation from mosquito specimens, and cytopathic effect was observed following inoculation into BHK-21 and C6/36 cells. The YNDC55-33 virus isolate was successfully amplified with the flavivirus primes, and a long sequence containing 300 nucleotides was obtained. Following sequence alignment using the BLAST tool, the sequence of the YNDC55-33 virus isolate had high homology with that of the genotype I Japanese encephalitis virus. A long sequence with 10 845 nucleotides in length, which encoded 3 432 amino acids, was obtained by splicing the full sequence of the YNDC55-33 virus isolate. Phylogenetic analysis based on the whole-genome sequence and E gene sequence of the YNDC55-33 virus isolate showed that the new YNDC55-33 virus isolate was most closely related to the genotype I Guizhou isolate (GenBank accession number: HM366552), with nucleotide homology of 98.5% and amino acid homology of 99.4%, and the YNDC55-33 virus isolate shared 97.96% ± 0.33% nucleotide homology and 99.35% ± 0.08% amino acid homology with other genotype I Japanese encephalitis virus isolates, and < 90% nucleotide homology and < 98% amino acid homology with other genotypes of Japanese encephalitis virus. The YNDC55-33 virus isolate and the live attenuated virus vaccine candidate SA14-14-2 isolate differed at 16 amino acid sites on E gene, and 7 out of 8 key amino acid sites related to neurovirulence. The secondary and tertiary structures of the E protein of the YNDC55-33 virus isolate were predicted to be characterized by random coils. Conclusions A genotype I Japanese encephalitis virus was isolated from Cx. tritaeniorhynchus in Dongchuan District, Kunming City. This virus isolate and the live attenuated virus vaccine candidate SA14-14-2 isolate does not differ at antigenic epitopes-related key amino acid sites, and the major protein structure of the virus isolate is random coils. This study adds new data for the epidemiological distribution of Japanese encephalitis virus in Yunnan Province, which may provide insights into the prevention and control of Japanese encephalitis in the province.

Objective:To explore the high-risk factors for parenteral nutrition associated cholestasis（PNAC）in extremely/ultra-low birth weight infants，and establish a risk Alignment Diagram prediction model.Methods:We retrospectivly analyzed the clinical data of hospitalized extremely/ultra-low birth weight infants admitted to Neonatology Department at Quanzhou Children's Hospital from January 2019 to December 2020，using multivariate Logistic regression analysis to screen for independent risk factors for the occurrence of PNAC.An Alignment Diagram model prediction model for PNAC was constructed by using R software，and the performance of the model was evaluated through receiver operating characteristic curves.Results:A total of 203 extremely/ultra-low birth weight infants were included，with a median gestational age of 29.14（28.00，30.86）weeks and a median birth weight of 1　170（1　000,1　300）g.Among them，26（12.81%）cases developed PNAC.Multivariate Logistic regression analysis showed that the duration of parenteral nutrition（ OR=1.015 ，95% CI 1.003-1.034），the cumulative amount of glucose（ OR=1.014 ，95% CI 1.001-1.028），small for gestational age（ OR=3.455 ，95% CI 1.127-10.589），and neonatal sepsis（ OR=3.142 ，95% CI 1.039-9.503）were independent risk factors for PNAC（ P＜0.05）；The four independent risk factors mentioned above were introduced into R software to construct an Alignment Diagram model，the area under the receiver operating characteristic curve was 0.835（95% CI 0.842-0.731），and the results of the Hosmer Limeshow goodness of fit test show that：χ 2=5.34，degree of freedom=8， P=0.72.A calibration curve indicated good consistency between the predicted probability of the model and the actual occurrence rate，with good accuracy. Conclusion:The Alignment Diagram model constructed based on four independent risk factors of the duration of parenteral nutrition，glucose accumulation，small for gestational age infants，and neonatal sepsis exhibits high predictive ability，and is expected to provide an intuitive and convenient visualization tool for preventing or reducing the occurrence of PNAC in extremely/ultra-low birth weight infants

There are millions of patients with taxane-induced peripheral neuropathy （TIPN）， and there is no effective treatment or prevention measure in clinical practice. The occurrence of TIPN may be related to the dosage form of paclitaxel drugs， genetic and molecular markers， drug dosage and chemotherapy cycle， patient factors， etc. At present， drugs for treating TIPN mainly include those that inhibit axonal degeneration （such as dosazosin， tamsulosin）， prevent mitochondrial dysfunction （such as glutathione trisulfides， antioxidants α -lipoic acid）， improve calcium imbalance in the internal environment （Shaoyao gancao decoction， N-type voltage-gated calcium channel inhibitor IPPQ）， and inhibit neuroinflammation （such as chemokine inhibitors and selective interleukin-8 receptor inhibitors DF2726A）. Further exploration of drug treatment strategies targeting different induction mechanisms is expected to become a new direction for precise clinical prevention and personalized treatment of TIPN.

Primary central nervous system lymphoma (PCNSL) is a highly aggressive malignant lymphoma. As most chemotherapy drugs have difficulty in crossing the blood-brain barrier, PCNSL shows a difficulty in clinical treatment, a high recurrence rate and a poor prognosis. Early identification of relapsed patients and prompt initiation of salvage therapy play a critical role in the improvement of patients' prognosis. Brain biopsy is the gold standard to identify recurrence, while the risk of operation failure and complications is still high. Non-invasive imaging techniques are beneficial for early identification of recurrence in PCNSL and can provide an important basis for guiding relapsed patients to adjust treatment plans in time. However, there is no unified evaluation standard for imaging methods of monitoring the relapsed lesions of PCNSL. With the further research of the pathophysiological mechanism of PCNSL, biomarker detection has become a new method to identify recurrence and more clinical evidence is still needed in the future.

Objective:To explore the medication law of wind drugs in Pi Wei Lun; To provide reference for the clinical application of wind drugs in the diagnosis and treatment of diseases. Methods:Through searching the prescriptions with wind drugs in Pi Wei Lun, wind drugs and the dosages were extracted. SPSS 25.0 software was used for descriptive statistics of wind drugs data. Traditional Chinese Medicine Inheritance Calculation Platform V3.0 was used for frequency statistics, dosage statistics and association rule analysis of Chinese materia medica. Results:Totally 40 prescriptions were included (40/61, 65.57%), involving 14 kinds of Chinese materia medica, mainly including Cimicifugae Rhizoma, Bupleuri Radix, Notopterygii Rhizoma et Radix, Saposhnikoviae Radix, Zingiberis Rhizoma Recens and so on. The ancient dosage of wind drugs was 0.68 g to 4.00 g, and the maximum was 8 g and the minimum was 0.09 g. However,the modern dosage was 3-10 g. The dosage ratio of commonly used of wind drugs was 3%-60%, and its efficacy varied. Through the analysis of association rules algorithm, the study gained 25 medicinal pairs, and the group Cimicifugae Rhizoma-Bupleuri Radix acquired the highest supporting degree. Under the conditions of a confidence level of 75% and support levels of 10%, 15%, and 20%, there were 14, 7, and 3 drug association rules, respectively. The efficacy of commonly used wind drugs varied depending on their ratio.Conclusion:Li Dongyuan attached great importance to develop and ascend the yang of spleen so that he used wind drugs more often for ascending yang, supplementing the original qi, dispelling wind, dispelling dampness, dissipating heat, resolving depression, purging the liver, relaxing the bowels, unblock the orifices and directing meridians.

Objective To investigate whether lidocaine can reverse Kupffer cell dysfunction in diabetic mice, as well as the mechanism of lidocaine in affecting liver abscess formation by improving the phagocytic function of Kupffer cells. Methods C57BLKS/J db/db mice were divided into diabetes control group and diabetes+lidocaine group, and C57BLKS/J db/m mice were divided into non-diabetes control group and non-diabetes+lidocaine group, with 5 mice in each group. All mice were fed with the suspension of Klebsiella pneumoniae . Kupffer cells were collected from each group and were cultured in vitro; an electron microscope was used to measure the change in ultrastructure, and Kupffer c ells were measured in terms of the levels of inflammatory mediators, the expression level of intercellular adhesion molecule-1 (ICAM-1), the chemotactic function of neutrophils, and phagocytic function; liver abscess formation was also observed. The Kruskal-Wallis H test was used for comparison of continuous data between multiple groups, and the Mann-Whitney U test was used for further comparison between two groups; the chi-square test was used for comparison of categorical data between groups. Results Compared with the non-diabetic mice, the diabetic mice had significant reductions in mitochondria and rough endoplasmic reticulum, endoplasmic reticulum dilation, mitochondrial swelling, and an increase in lipid droplets in Kupffer cells. Compared with the non-diabetes control group, the diabetes control group had significant increases in the levels of nitric oxide (NO) (4.95±0.06 μmol/L vs 1.34±0.13 μmol/L, P < 0.05), interleukin-6 (IL-6) (740.04±8.58 pg/mL vs 515.77±4.62 pg/mL, P < 0.05), tumor necrosis factor-α (TNFα) (774.23±7.98 pg/mL vs 461.51±1.76 pg/mL, P < 0.05), interferon gamma (IFNγ) (842.33±14.79 pg/mL vs 542.47±6.75 pg/mL, P < 0.05), and ICAM-1 (2.40±0.02 vs 1.33±0.01, P < 0.05) in Kupffer cells, a significant increase in neutrophil chemotaxis (100.80±10.18 vs 13.80±3.70, P < 0.05), and a significant reduction in phagocytic capacity (9.86±1.82 vs 60.00±3.54, P < 0.05), with no effect on liver abscess formation (40% vs 0, P > 0.05). Compared with the diabetes control group, the diabetes+lidocaine group had significant reductions in the levels of NO (3.35±0.28 μmol/L vs 4.95±0.06 μmol/L, P < 0.05), IL-6 (688.42±36.34 pg/mL vs 740.04±8.58 pg/mL, P < 0.05), TNFα (631.15±4.30 pg/mL vs 774.23±7.98 pg/mL, P < 0.05), IFNγ (704.56±3.64 pg/mL vs 842.33±14.79 pg/mL, P < 0.05), and ICAM-1 (1.50±0.02 vs 2.40±0.02, P < 0.05) in Kupffer cells, a significant reduction in neutrophil chemotaxis (33.40±5.60 vs 100.80±10.18, P < 0.05), and a significant increase in phagocytic capacity (49.20±2.59 vs 9.86±1.82, P < 0.05), with no effect on liver abscess formation (0 vs 40%, P > 0.05). Conclusion Lidocaine can inhibit Kupffer cell inflammatory response and improve the phagocytic function of Kupffer cells in diabetic mice, thereby exerting a protective effect on Kupffer cells, but it had no effect on liver abscess formation.

Objective To analyze the loss coefficient of bivalent live attenuated oral polio vaccine（bOPV) in Hongshan District, and understand the causes and influencing factors, and to provide a basis for scientifically formulating vaccine use plans and regulating vaccine management. Methods Using the data of Hubei Province Immunization Planning Information System and Hongshan District Vaccination Storage and Management System, a special questionnaire was designed to understand the vaccination and use of bOPV in various immunization units in Hongshan District. A descriptive statistical analysis method was used to calculate the loss coefficient. Results The bOPV loss coefficient of 25 vaccination units in Hongshan District was 1.69. The difference of the loss coefficient between the out-patient clinics of the District Health and Family Planning Committee（1.35）and the community clinics undertaken by large hospitals (2.01）was statistically significant. The difference of the loss coefficient between the inoculation once a week（1.57）and 2-6 times per week（1.94）was statistically significant. In terms of vaccine-disabled time, the two groups of <4 hours and >4 hours had different calculated loss coefficients 1.86 and 1.61, respectively, and the difference was statistically significant. Multivariate regression analysis of the factors related to the loss coefficients found that different outpatient attributes had a greater impact on the loss coefficient than the inoculation cycle and the number of inoculation stations. The loss coefficient for 2016 and 2017 was 2.09 and 1.75, respectively. The difference was statistically significant. Conclusion The management and use of live attenuated bivalent polio vaccine in Hongshan District was relatively standardized. The vaccine loss can by further reduced by strengthening supervision and assessment, setting up centralized vaccination and standardizing publicity and training.

Objective To explore the relationship between sudomotor function and peripheral artery disease (PAD),and to evaluate the effectiveness of sudomotor function testing to screen diabetic patients at high risk of PAD comparing to ankle-brachial index (ABI) and toe-brachial index (TBI).Methods A total of 263 diabetes mellitus (DM) outpatients in the 306th Hospital of PLA from August 2014 to April 2015 were enrolled in the study.ABI and TBI were measured by the Doppler method.Sudomotor function was evaluated by measuring the electrochemical skin conductance (ESC) of the hands and feet using the Sudoscan instrument.Cardiovascular autonomic neuropathy (CAN) was assessed and recorded as cardiac autonomic neuropathy risk-score (CAN-RS) by Sudoscan.Results ESC values of the feet and hands were positively correlated with ABI and TBI.The diabetes patients with abnornal ABI and TBI had significantly lower hand ESC [(45.63±12.87) μS vs.(68.10±17.40) μS,(59.17±19.58) μS vs.(68.57±17.11) μS;P＜ 0.05] and feet ESC [(44.54±25.48) μS vs.(70.92±19.46) μS,(59.21±24.52) μS vs.(71.71±19.02) μS;P＜ 0.05],and higher CAN-RS[(49.17± 15.41)％ vs.(36.33±16.25)％,(44.90±16.09)％ vs.(35.39±16.05)％;P＜ 0.05],than diabetes patients with normal ABI and TBI.Using ABI as the gold standard,the areas under the receiver operating characteristics (ROC) curve of the diagnostic performance of hands ESC,feet ESC and CAN-RS to identify PAD were 0.87,0.84 and 0.74,respectively (P＜0.001).Conclusion Sudomotor function testing can be helpful and beneficial to identify PAD in patients with diabetes.

Objective To study the compliance of medical care at public hospitals in China with corresponding clinical pathway for inpatients of community acquired pneumonia ( CAP) . Methods Thirty medical records of inpatients with CAP admitted in 2014 were randomly sampled from 18 public hospitals in Shanghai, Hubei province and Gansu province, for assessment on their compliance with corresponding clinical pathways. Results The average compliance rate of inpatient care of CAP at public hospitals was 65. 1%. The study the lowest compliance rates in both timely examinations/diagnosis and severity assessment within three days after admission (0. 7% and 26. 6%, respectively). The compliance rate of reasonable initial antibiotic use was 48. 5%. Conclusions The compliance rate of inpatient care of CAP is low in China. Thus admission examinations, patient severity assessment and antibiotic utilization should be enhanced accordingly for their compliance.

Objective To understand the virus-carrying status and infection condition of Culicoides in Yunnan province.Methods Culicoides, cattle serum samples and pig serum samples were collected in Qujing City in Yunnan province from Sep.to Nov.in 2011; the supernatant of Culicoides was inoculated in C6/36 cells to isolate virus.Suspected isolates were identified by molecular biology techniques and titers of virus antibodies in pigs and cattles serum samples were tested by VN.Results 8 300 short tarsal Culicoides (8 300/18 160) and 7 100 Ryukyu Culicoides (7 100/18 160) were identified among 18 160 Culicoides specimens.Two suspected virus strains were isolated and designated as SC093 and SC233.The nucleotide sequence homology of VP2, VP4 and VP8 sequences with Banna virus Chinese strain ( Accession number:AF134526) is up to 99%.1 out of 200 (1/200) cattle serum samples was antibody positive against Banna virus with 0.5%positive rate.And the neutralizing antibody titers of SC093 and SC233 with above Banna virus antibody positive cattle serum were 160 and 40.10 out of 535 ( 10/535 ) pig serum samples were antibody positive against Banna virus with 1.7%positive rate.And the neutralizing antibody titers of SC093 and SC233 with above Banna virus antibody positive pig serum samples were 80-320 and 20-80 respectively.Conclusions The SC093 and SC233 virus strains isolated from Shizong were identified to be Banna virus and it could cause infection in pigs and cattles.This is the first report that Banna virus was isolated from Culicoides.