Anthocyanidin reductase (ANR) is one of the key enzyme in the flavonoid biosynthetic pathway, and its catalytic activity is important for the synthesis of plant anthocyanin. In this study, specific primers were designed according to the transcriptome data of Lonicera japonica Thunb., and the CDS, gDNA and promoter sequences of ANR genes from Lonicera japonica Thunb. and Lonicera japonica Thunb. var. chinensis (Wats.) Bak. were cloned. The results showed that the CDS sequences of LjANR and rLjANR were 1 002 bp, the gDNA sequences were 2 017 and 2 026 bp respectively, and the promoter sequences were 1 170 and 1 164 bp respectively. LjANR and rLjANR both contain 6 exons and 5 introns, which have the same length of exons and large differences in introns. The promoter sequences both contain a large number of light response, hormone response and abiotic stress response elements. Bioinformatics analysis showed that both LjANR and rLjANR encoded 333 amino acids and were predicted to be stable hydrophobic proteins without transmembrane segments and signal peptides. The secondary structures of LjANR and rLjANR were predicted to be mainly consisted of α-helix and random coil. Sequence alignment and phylogenetic analysis showed that LjANR and rLjANR had high homology with Actinidia chinensis var. chinensis, Camellia sinensis and Camellia oleifera, and were closely related to them. The expression levels of LjANR and rLjANR were the highest in flower buds and the lowest in roots. The expression patterns at different flowering stages were similar, with higher expression levels in S1 and S2 stages and then gradually decreased until reaching the lowest level in S4 stage, after a slow increase in S5 stage, the expression levels decreased again. The expression levels of ANR genes in the two varieties showed significant differences in roots, S2 and S5 stages, while the differences in stems, flower buds, S1, S3 and S6 stages were extremely significant. The prokaryotic expression vector pET-32a-LjANR was constructed for protein expression. The target protein was successfully expressed of about 59 kD. This study lays a foundation for further study on the function of ANR gene and provides theoretical guidance for breeding new varieties of Lonicera japonica Thunb.

Abstract: Objective　To analyze the emergency response and long-term intervention effects after the detection of infectious snails epidemic by loop-mediated isothermal amplification (LAMP) assays in Hannan District, Wuhan City, and to explore the application of LAMP in early surveillance and early-warning of schistosomiasis transmission. Methods Snails picked up by the risk monitoring system in Hannan District were examined by anatomical microscopy and LAMP technology to identify the schistosomiasis infection. Emergency response and intensive intervention were initiated in the environment where positive snails appeared, and the long-term effects were evaluated. Results In May 2018, the infectious snails were detected by LAMP technology in Hannan District, and the positive snails were located in Zhujiacha, Dongzhuang Village, Obstacles and weeds were removed and buried by machine in Zhujiacha. 12 700 m2 of snails were killed by drugs, and the mortality rate of snails was more than 80%; no new seropositive persons were found in the emergency examination within 500 m of the positive snail sites. 506 people were examined in Dong Zhuang Village at the end of the year, and 30 positive IHA cases were detected with a blood positive rate of 5.93%, no positive fecal test was found, and all positive blood test patients took preventive medication. The monitoring results of sentinel rats and wild feces were all negative. Health education was carried out, 7 warning signs were deployed and refreshed, and 500 publicity brochures were distributed. After nearly three years of intensified intervention and monitoring in the villages where the positive environment is located, and the density of snails on the stubborn snail has dropped from 0.094/frame to 0.027/frame, and the positive rate of blood test in Dongzhuang Village has steadily dropped from 5.93% to 3.74%. Conclusions The infected snails missed by microscopy were detected by LAMP in Hannan District, which created conditions for the rapid emergency treatment of environment and elimination of positive snail and improved the sensitivity of the surveillance and early warning system in transmission-interrupted areas.

Objective:To explore the clinical efficacy of adult donor dual kidney transplantation.Methods:Retrospective analysis of case data of 13 adult donor kidney dual kidney transplantation (DKT) performed in the The Second Xiangya Hospital of Central South University from September 2016 to December 2020. For 13 donors, the average age and BMI were (53.5±12.4)years and (24.3±2.8) kg/m 2, respectively. Their mean Serum creatinine (SCr) at admission and before procurement was (132.9±54.1)and (228.7±112.4)μmol/L, respectively. 3 of them had diabetes mellitus history, and 8 had hypertension history. 11 met the United Network for Organ Sharing (UNOS) DKT criteria and 6 met Remuzzi score DKT criteria. For 13 recipients, the average age and BMI were (39.3±8.9)years and (20.2±2.4)kg/m 2, respectively. All of them received ABO blood type-matched kidney transplants. 2 of them had their grafts transplanted in the bilateral iliac. In 12 cases, the grafts filled rapidly and urinated immediately when opening blood flow. In 1 case, the grafts were dark in color and vascular showed weak pulsation after opening blood flow. The time to recovery of perioperative graft function (from the day of surgery to the natural reduction of SCr to the normal range 44-133μmol/L), the occurrence of delayed graft function (DGF), acute rejection (AR), ureteral and surgical incision complications, as well as the recipients’ final follow-up SCr, eGFR, urinary protein, and grafts outcome were observed. Risk factors affecting outcomes were assessed by univariate logistic regression analysis. Results:The SCr dropped to the normal range at discharge in 10 recipients, and the average recovery time was (13.8±13.0) days. In other 3 cases SCr at discharge were 300.0, 149.0, 152.5μmol/L. 4 cases had DGF, 4 had AR, 1 experienced urinary fistula, and 1 experienced incisional dehiscence, which were treated with anti-rejection, J-tube implantation, continuous catheterization to maintain bladder void, secondary suturing, respectively. The follow-up time ranged from 4 to 54 months, with a median of 28(15.5, 31.0) months. At the final follow-up time, 10 cases had good graft function, 2 suffered impaired kidney function, and 1 experienced graft failure. The average SCr and eGFR except for graft failure patient were (144.2±101.3)μmol/L and (52.9±21.2)ml/min, respectively. 4 had positive urine protein. Univariate logistic regression analysis showed that donor age, BMI, history of diabetes mellitus and hypertension, and SCr were not significantly correlated with recipients’ DGF and graft impairment ( P>0.05), and due to the small sample size, multifactorial logistic regression analysis was not performed. Conclusion:The short to medium-term effects of adult donor DKT coule be safe and feasible.

Objective To understand and master the infection status of key parasitic diseases in Baise City from 2006 to 2015, so as to provide evidence for the development and adjustment of the control strategy for parasitic diseases. Methods The relevant information of the final evaluation of the key parasitic diseases in 12 counties (cities and districts) of Baise City from 2006 to 2015 was collected, and analyzed. Results During the period of 2006-2015, a total of 20 654 person-times were investigated for parasitic diseases in Baise City and 1 147 persons were infected, with an average infection rate of 5.55%. In 2006, the infection rate was the highest (28.67%, 362/1 254), while the lowest was in 2013 (2.08%, 44/2 113). The infection rate of Clonorchis sinensis was the highest, and the infection rate of Ascaris lumbricoides was the second. The infection rates of the males and females were 5.35% (631/11 795) and 5.82% (516/8 859) respectively, with no significant difference between them (χ² = 2.175, P > 0.05). The highest infection rate existed in the 40-49 years group, the rural residents, the farmers and the crowd with the educational level of senior high middle school or above. Conclusions The infection rate of parasitic diseases presents a decline trend overall in Baise City from 2006 to 2013, however it rebounded in 2014 and 2015. The next step is to strengthen the prevention and control of parasitic diseases by promoting health education, regular insect repellent, and fecundity management.

Near infrared spectroscopy combined with chemometrics methods was used to distinguish Ganoderma lucidum samples collected from different origins, and a prediction model was established for rapid determine polysaccharides contents in these samples. The classification accuracy for training dataset was 96.87%, while for independent dataset was 93.33%; as for the prediction model, 5-fold cross-validation was used to optimize the parameters, and different signal processing methods were also optimized to improve the prediction ability of the model. The best square of correlation coefficients for training dataset was 0.965 4, and 0.851 6 for validation dataset; while the root-mean-square deviation values for training dataset and validation dataset were 0.018 5 and 0.023 6, respectively. These results showed that combining near infrared spectroscopy with suitable chemometrics approaches could accuracy distinguish different origins of G. lucidum samples; the established prediction model could precious predict polysaccharides contents, the proposed method can help determine the activity compounds and quality evaluation of G. lucidum.
Fungal Polysaccharides ; analysis ; Geography ; Least-Squares Analysis ; Reishi ; chemistry ; Spectroscopy, Near-Infrared

Objective To evaluate the monitoring results of medium-and long-term program of schistosomiasis control in Baise City,so as to provide the reference for the elimination of schistosomiasis.Methods The data of schistosomiasis control in Baise City from 2004 to 2015 were collected and analyzed.Results By the end of 2015,the total number of regular screening serum tests for schistosomiasis in the planning period was 10 244 person-times,with 649 positive cases.The number of feces tests was 2 158 person-times in the permanent resident population,and the number was 2 683 person-times in the floating popu-lation.The Oncomelania hupensis snail survey area was 150.04 hm2,and the accumulated snail control area was 2.03 hm2.No schistosomiasis patients or schistosome-infected snails were found.Conclusion The effect of medium-and long-term program of schistosomiasis control is effective in Baise City,and the criterion of schistosomiasis elimination has been achieved.

Objective To investigate the epidemiology,diagnosis and treatment of an imported falciparum malaria patient in Xilin County,Baise City,so as to provide the reference for improving the diagnosis and treatment of falciparum malaria pa-tients in the future. Methods The epidemiological and clinical data were collected and analyzed. Results The patient had lived in Africa where the malaria was epidemic. The disease attacked him after his coming back home from abroad. The blood test for Plasmodium falciparum was positive. In Xilin County,no local Plasmodium infection was found from 2004 to 2016,and therefore,we concluded that this case was overseas imported. Conclusion The monitoring of overseas returnees in Xilin Coun-ty should be strengthened to timely diagnose and treat the imported cases of malaria.

AIM:To investigate the value of 3D-optical coherence tomography (OCT) in the diagnosis of macular disease before phacoemulsification.METHODS:Clinical records of 423 cataract patients (512 eyes) who underwent phacoemulsification combined with intraocular lens implantation in our hospital from June to December in 2015 were retrospectively analyzed.In addition to preoperative routine examination of fundus, Topcon 3D-OCT 2000 was used to examine the macula, the detection rate of macular disease was compared, risk factors of cataract combined with macular disease were analyzed.RESULTS:The OCT image results of 305 cases (384 eyes, 72.1%) were successfully obtained, 133 cases showed macular disease (146 eyes), the detection rate was 28.5% (95%CI:27.64%-29.40%);the macular disease of 35 cases (37 eyes) were detected by routine examination of fundus before operation, the detection rate was 7.2% (95%CI:6.72%-7.74%);the detection rate of 3D-OCT was significantly higher than routine examination of fundus for macular disease (χ2=79.05, P<0.01).Female, over 65 years old, surgical history of diseased eye, and high myopia were risk factors of cataract combined with macular disease, the relative risk was 1.705 (95%CI:1.091,2.664), 1.893 (95%CI:1.219,2.939), 6.593 (95%CI:2.027,21.447) and 95%CI:5.130 (2.841,9.263) respectively, the risk of cataract combined with macular disease showed an increasing trend with rising age.CONCLUSION:In preoperative examination of cataract patients, 3D-OCT has higher sensitivity in the detection of macular disease, especially for women, over 65 years old, high myopia and surgical history of diseased eye, 3D-OCT can be used as a routine preoperative examination.

Objective To investigate the anti-hyperglycemic effect of the Chinese medicine ingredients such as berberine,baicalin,puerarin and liquiritin on the optimal insulin resistance (IR)-HepG2 cell model by oleic acid.It would provide the theoretical basis for the optimization of Chinese medicine prescription or anti-hyperglycemic components combination.Methods Different concentrations (0.1,0.2,0.5,and 1.0 mmol/L) of oleic acid were used to induce HepG2 cells for different time (24,36 and 48 h),the glucose consumption was measured byglucose oxidase assay,and cell viability was detected by CCK-8 assay to define the optimal inducing concentration and time for IR-HepG2 cell model.Then the cell morphological changes were detected by oil red O staining.Finally,the stability of IR-HepG2 cell model was tested.After the IR-HepG2 model was optimally established,the glucose consumption,glycogen content and cell viability were detected after 24 h administration with different concentrations of berberine,baicalin,puerarin and liquiritin by anthrone method,glycerol phosphate oxidase assay and CCK-8 assay respectively.Results The optimal oleic acid-induced concentration was 1 mmol/L and the optimal induced time was 24 h for the IR-HepG2 cell model that could keep stable more than 36 h.Comparing with IR model group,berberine,puerarin and baicalin significantly increased the glucose consumption,whereas liquiritin did not show significant change in the glucose consumption except for 1 μmol/L.Only 160 μmol/L puerarin and 1 μmol/L baicalin significantly inhibited IR-HepG2 cell viability.Moreover,berberine,puerarin,and baicalin significantly elevated the glycogen content;Liquiritin did not change glycogen content significantly.Conclusion The IR-HepG2 cell model could be stably established with 24 h treatment of 1 mmol/L oleic acid.Berberine,puerarin,and baicalin significantly increased the glucose consumption and glycogen content in the IR-HepG2 cells.The results suggest that berberine,baicalin and puerarin maybe perform different pathways of anti-hyperglycemic effects due to different incentives ofIR.

Objective To investigate the effects of IL-21,IL-23 and IL-4 in the pathogenesis of Guillain-Barré syndrome(GBS) of children,and to analyze the influence of intravenous immunoglobulin(IVIG) on the serum levels of IL-21,IL-23 and IL-4 in GBS patients.Methods Forty-one pediatric patients with GBS hospitalized in our department from Jan.2005 to Sep.2012 were studied.According to the time of IVIG administration,patients were divided into group A and group B,given IVIG respectively within the 7th day and the 8th-12th day.According to Hughes score,patients were divided into mild group and severe group.The serum levels of IL-21,IL-23 and IL-4 were detected by means of ELISA before and after treatment.Results Before treatment,the serum levels of IL-21,IL-23 and IL-4 had no signifiant difference between group A and group B.After treatment,the serum levels of IL-21,IL-23 in group A and group B were significantly decreased than those before treatment (all P ＜ 0.05),with no significant difference in IL-4 levels.There was no signifiant difference in group A and group B in the serum levels of IL-21,IL-23 and IL-4 after treatment (all P ＞0.05).In the severe group,the serum levels of IL-21,IL-23 were significantly higher than those in the mild group before and after treatment (all P ＜ 0.05),with no significant difference in IL-4 levels.After treatment,the serum levels of IL-21,IL-23 were significantly decreased than those before treatment in the mild group and the severe group (all P ＜ 0.05),with no significant difference in IL-4 levels.Conclusions The serum levels of IL-21,IL-23 are significantly increased in GBS children,indicating they play an important role in the pathogenesis of GBS.Levels of IL-21 and IL-23 are significantly decreased after administration of IVIG,which showed that IVIG could inhibit the secretion of IL21 and IL-23,reduce cellular inflammatory response,and eventually prevent the development of GBS.IL-4 secretion is not obviously affected with IVIG,it might have a role in the recovery of GBS.