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 possible mechanism of Xiebaisan in protecting against allergic asthma in rats from the perspective of host intestinal flora metabolism.Methods SPF SD rats were divided into normal group(NC group),model group(M group),and Xiebaisan group.The allergic asthma rat model was established by ovalbumin.Changes in lung histopathology were observed by HE staining.Colon contents were harvested for 16S rDNA high-throughput sequencing to assess changes in the intestinal flora structure and function.Serum and lung tissue samples were collected for non-targeted metabolomics by Ultra-high performance liquid-time-of-flight mass spectrometer.Results HE staining showed some improvement of lung histomorphology in asthmatic rats in the Xiebaisan group compared with that in the M group.16S rDNA high-throughput sequencing showed that the diversity of intestinal flora was decreased in the M group and increased in the Xiebaisan group compared with the M group,the microecosystem of intestinal was improved.Non-targeted metabolomics of serum showed regulation of amino acid metabolism and the mTOR pathway in the Xiebaisan group,and partially reversed differential metabolite expression in the M group.Non-targeted metabonomics of lung tissue samples showed regulation of carbon metabolism,vascular smooth muscle and cAMP signaling pathways in the Xiebaisan group,and partially reversed differential metabolite expression in the M group.Conclusions The protective effects of Xiebaisan on allergic asthma in rats may be related to improvement of the morphological structure of lung tissue,the diversity of intestinal flora,and regulation of mTOR,vascular smooth muscle contraction,and cAMP pathways,which affect amino acid and carbon metabolism.

Objective:To investigate the application effect of the compound flap transfer in tubularized incised plate (TIP) urethroplasty.Methods:Children with hypospadias who received TIP in the Department of Pediatric Urology, the Affiliated Xuzhou Children’s Hospital of Xuzhou Medical University from January 2018 to March 2022 were prospectively divided into compound flap transfer group (group A) and traditional TIP group (group B) by random number method. Group A was treated with TIP modified by compound flap transfer, and group B was treated with traditional TIP covered with fascia.The improvement of the compound flap transfer for TIP operation is mainly reflected in the multi-layer cover of the new urethra and the free skin embedded in the incision of the penile head. According to the intention-to-treat (ITT) principle, the final full analysis set (FAS) includes both group A and group B. Some patients were excluded from the analysis of the primary endpoint events due to reasons such as loss to follow-up or treatment group switch. Additionally, the final per-protocol set (PPS) consisting of group A and group B, which adheres to the study protocol, is subjected to statistical analysis.Independent sample t-test or Wilcoxon rank sum test was used for average age, the width of the penile head, degree of chordee, length of neourethra, and operative time. The classification of hypospadias, proportion of dorsal tunica albuginea plication, and incidence of postoperative complications were compared using the Chi-square test or Fisher’s exact test, P < 0.05 was considered statistically significant. Results:According to ITT principles, 50 children were included in FAS group A, ranging in age from 8 months to 15 years and 2 months, with an average age of 4 years. Group B included 50 children, ranging in age from 10 months to 14 years and 9 months, with an average age of 4 years and 1 month. Thirty-seven children in PPS group A were included, ranging in age from 1 year 2 months to 12 years 1 month, with an average age of 4 years. Group B consisted of 41 children, ranging in age from 1 year 2 months to 11 years 9 months, with an average age of 4 years 2 months. Statistical analysis showed that no matter FAS set or PPS, there were no statistically significant differences in the mean age, hypospadias type, average penile head width, average penile subcurvature number, number of dorsal tunica albuginea plication, and the average length of plastic neourethra between group A and group B ( P > 0.05). The average operative time of group A was higher than that of group B. The difference was statistically significant ( P < 0.05). The postoperative follow-up was 3 to 48 months, with an average follow-up of 2 years and 3 months. The complication rate of group A was lower than that of group B [10.81% (4/37) vs 29.27% (12/41)], and the difference was statistically significant ( P< 0.05). Urethral fistula occurred in 3 cases (8.11%), respectively and 6 cases (14.63%) in the two groups, the difference was not statistically significant ( P>0.05); urethral stricture occurred in 1 case (2.70%) and 5 cases (12.20%) respectively, the difference was not statistically significant ( P>0.05), There were 0 case and 1 case (2.44%) of urethral orifice descending or urethral dehiscence respectively, and the difference was not statistically significant ( P> 0.05). Conclusion:TIP with an improved compound flap transfer can reduce the overall postoperative complication rate and is worthy of promotion.

Objective:To investigate the application effect of the compound flap transfer in tubularized incised plate (TIP) urethroplasty.Methods:Children with hypospadias who received TIP in the Department of Pediatric Urology, the Affiliated Xuzhou Children’s Hospital of Xuzhou Medical University from January 2018 to March 2022 were prospectively divided into compound flap transfer group (group A) and traditional TIP group (group B) by random number method. Group A was treated with TIP modified by compound flap transfer, and group B was treated with traditional TIP covered with fascia.The improvement of the compound flap transfer for TIP operation is mainly reflected in the multi-layer cover of the new urethra and the free skin embedded in the incision of the penile head. According to the intention-to-treat (ITT) principle, the final full analysis set (FAS) includes both group A and group B. Some patients were excluded from the analysis of the primary endpoint events due to reasons such as loss to follow-up or treatment group switch. Additionally, the final per-protocol set (PPS) consisting of group A and group B, which adheres to the study protocol, is subjected to statistical analysis.Independent sample t-test or Wilcoxon rank sum test was used for average age, the width of the penile head, degree of chordee, length of neourethra, and operative time. The classification of hypospadias, proportion of dorsal tunica albuginea plication, and incidence of postoperative complications were compared using the Chi-square test or Fisher’s exact test, P < 0.05 was considered statistically significant. Results:According to ITT principles, 50 children were included in FAS group A, ranging in age from 8 months to 15 years and 2 months, with an average age of 4 years. Group B included 50 children, ranging in age from 10 months to 14 years and 9 months, with an average age of 4 years and 1 month. Thirty-seven children in PPS group A were included, ranging in age from 1 year 2 months to 12 years 1 month, with an average age of 4 years. Group B consisted of 41 children, ranging in age from 1 year 2 months to 11 years 9 months, with an average age of 4 years 2 months. Statistical analysis showed that no matter FAS set or PPS, there were no statistically significant differences in the mean age, hypospadias type, average penile head width, average penile subcurvature number, number of dorsal tunica albuginea plication, and the average length of plastic neourethra between group A and group B ( P > 0.05). The average operative time of group A was higher than that of group B. The difference was statistically significant ( P < 0.05). The postoperative follow-up was 3 to 48 months, with an average follow-up of 2 years and 3 months. The complication rate of group A was lower than that of group B [10.81% (4/37) vs 29.27% (12/41)], and the difference was statistically significant ( P< 0.05). Urethral fistula occurred in 3 cases (8.11%), respectively and 6 cases (14.63%) in the two groups, the difference was not statistically significant ( P>0.05); urethral stricture occurred in 1 case (2.70%) and 5 cases (12.20%) respectively, the difference was not statistically significant ( P>0.05), There were 0 case and 1 case (2.44%) of urethral orifice descending or urethral dehiscence respectively, and the difference was not statistically significant ( P> 0.05). Conclusion:TIP with an improved compound flap transfer can reduce the overall postoperative complication rate and is worthy of promotion.

OBJECTIVE To study the absorbed components of Xiebai powder in blood. METHODS UPLC-Q-TOF-MS/MS method was adopted. SD rats were randomly divided into blank group and administration group ，with 10 rats in each group. Blank group was given water intragastrically ，and administration groups were given 2 g/mL（by the amount of crude drug ）Xiebai powder solution intragastrically. Administration volume was 11.3 mL/kg，twice a day for 3 days. One point five hours after last administration，blood was taken from the abdominal aorta of each rat ，the serum was processed to obtain the supernatant for analysis；the relevant data in positive and negative ion mode were collected ，and the absorbed components of Xiebai powder in blood were analyzed and identified by using self-built secondary mass spectrometry database and consulting the relevant literature. RESULTS Totally 17 components from Xiebai powder were identified ，among which 6 components came from sovereign Moru salba，7 from minister Cortex Lycii ，12 from assistant Glycyrrhiza uralensis ，i.e. kukoamine A ，chlorogenic acid ，tachiogroside B，astringin，neoglycyrrhizin，glycyrrhizin，azelaic acid ，isoglycyrrhizin，glycyroside，anthocyanin，sebacic acid ，parthenolide， anthocyanin，18β-glycyrrhetinic acid ，6-gingerol，palmitoamide，erucamide. These compounds were mainly flavonoids ，alkaloids and organic acids. CONCLUSIONS In this study ，17 absorbed components of Xiebai powder in blood are preliminarily determined，which are consistent with the effect of Xiebai powder. They may be the pharmacodynamic substances of Xiebai powder.

Objective:To compare the effectiveness of modified Sugita and modified Shiraki in the treatment of severely concealed penis in children.Methods:A retrospective study was carried out on 72 children with congenital severely concealed penis, who were treated in the Department of Urology, Xuzhou Children′s Hospital Affiliated to Xuzhou Medical University from September 2016 to June 2021.Among them, 32 cases were treated with modified Sugita (modified Sugita group) and 40 cases were treated with modified Shiraki (modified Shiraki group). There was no significant difference in the age, body mass and body mass index between the 2 groups (all P>0.05). The operation time, increased length of the exposed part of the penis after operation, score of parental satisfaction half a year after operation (obtained by using the Likert scale) and postoperative complications (classified according to the modified Clavien-Dindo classification) were compared between the 2 groups.Measurement data comparison between groups were performed by t test, counting data were analyzed by Chi- square test. Results:The operative time of the modified Sugita group and the modified Shiraki group were (50.00±8.03) min and (40.30±9.27) min, respectively.The operative time was significantly different between 2 groups ( t=4.107, P<0.05). The increased length of the exposed part of the penis after operation was (1.80±0.30) cm in the modified Sugita group and (1.90±0.33) cm in the modified Shiraki group, and no significant difference was found between the 2 groups ( P>0.05). Six months after operation, the parental satisfaction score was (4.60±0.56) points in the modified Sugita group and (4.60±0.59) points in the modified Shiraki group.There was no significant difference in the parental satisfaction score between 2 groups ( P>0.05). In 72 cases, only 5 cases had grade Ⅰ complications.Postoperative complications were also not significantly different between the 2 groups ( P>0.05). Conclusions:After treating severely concealed penis in children with modified Sugita and modified Shiraki, the penis has a good appearance.These two surgeries have high parental satisfaction and low complications.They are easy to implement in clinical practice.The operation time of modified Shiraki is shorter than that of modified Sugita.

Objective:To introduce the application of a modified scrotoplasty for children with webbed penis.Methods:Retrospective study was carried out.The treatment results of 58 children with webbed penis in Department of Urology, the Affiliated Xuzhou Children′s Hospital of Xuzhou Medical University from June 2012 to April 2018 were analyzed.A total of 26 children with webbed penis and phimosis were treated with circumcision combined with modified scrotoplasty as modified scrotoplasty group, and 32 children with webbed penis and phimosis were treated with V-Y scrotoplasty as V-Y scrotoplasty group.The curative effect of webbed penis was compared between the two groups.Results:The operation time of V-Y scrotoplasty group and modified scrotoplasty group were (26.0±2.4) min and (28.0±3.2) min; the increased penis length in the horizontal position in the two groups were (0.30±0.06) cm and (0.40±0.06) cm; the score of parents′ satisfaction were (3.80±0.47) scores and (4.70±0.56) scores, there were significant differences in the average operation time, the increased penis length in the horizontal position and the score of parents′ satisfaction between the two groups ( t=2.703, 6.061, 6.652; all P<0.05). There was no significant difference in postoperative complications between the two groups ( P>0.05). Conclusions:Circumcision combined with modified scrotoplasty is used to repair webbed penis, which results in a good postoperative appearance, high parent satisfaction and easy clinical implementation.

Objective:To evaluate the effect of penile dorsal extension and bandaging after concealed penis surgery.Methods:In this study, 80 children who underwent concealed penile correction were randomly divided into a dorsal extension bandaging group (experimental group) and a traditional bandaging group (control group) from Xuzhou Children’s Hospital Affiliated to Xuzhou Medical University during September 2016 to September 2019. The control group was applied with traditional sleeve-type bandaging, and the test group was treated with penile dorsal extension bandaging. A total of 75 children was summarized in this study. Among them, there were 38 children in the experimental group, with a mean age of 64 months. Thirty-seven children were in the control group, with a mean age of 70 months. The incidence of complications during hospitalization, the number of calls to medical staff within the first 24 hours after surgery, the pain score of the child during dressing removal, and the time taken for dressing removal were compared and recorded. Measurement data were analyzed by t-tests and enumeration data were analyzed by chi-square tests between groups. All data were analyzed using software SPSS 17.0. Results:The incidence of complications during hospitalization was 5.26%(2/38) and 10.81%(4/37) in the experimental and control groups, respectively. This difference was not statistically significant( χ2=0.784, P>0.05). In the dorsal extension bandaging group and the traditional bandaging group, the average number of calls to medical staff 24 hours after surgery was (0.87 ± 0.91) and (1.54 ± 1.02) times, the difference was statistically significant ( t=2.996, P=0.003); as for the highest pain scores of the children when the dressing was removed were 5.21 ± 1.19 and 7.24 ± 1.20, the difference was statistically significant ( t=5.697, P< 0.001); the time taken to remove the dressing was (3.21 ± 1.24) min, (7.56 ± 1.88) min, and the difference was statistically significant ( t=11.917, P<0.001). Conclusions:The penile dorsal extension bandaging method after concealed penis surgery can reduce the number of calls to medical staff, reduce the time of dressing removal and the degree of pain in children.

Objective:To evaluate the effect of penile dorsal extension and bandaging after concealed penis surgery.Methods:In this study, 80 children who underwent concealed penile correction were randomly divided into a dorsal extension bandaging group (experimental group) and a traditional bandaging group (control group) from Xuzhou Children’s Hospital Affiliated to Xuzhou Medical University during September 2016 to September 2019. The control group was applied with traditional sleeve-type bandaging, and the test group was treated with penile dorsal extension bandaging. A total of 75 children was summarized in this study. Among them, there were 38 children in the experimental group, with a mean age of 64 months. Thirty-seven children were in the control group, with a mean age of 70 months. The incidence of complications during hospitalization, the number of calls to medical staff within the first 24 hours after surgery, the pain score of the child during dressing removal, and the time taken for dressing removal were compared and recorded. Measurement data were analyzed by t-tests and enumeration data were analyzed by chi-square tests between groups. All data were analyzed using software SPSS 17.0. Results:The incidence of complications during hospitalization was 5.26%(2/38) and 10.81%(4/37) in the experimental and control groups, respectively. This difference was not statistically significant( χ2=0.784, P>0.05). In the dorsal extension bandaging group and the traditional bandaging group, the average number of calls to medical staff 24 hours after surgery was (0.87 ± 0.91) and (1.54 ± 1.02) times, the difference was statistically significant ( t=2.996, P=0.003); as for the highest pain scores of the children when the dressing was removed were 5.21 ± 1.19 and 7.24 ± 1.20, the difference was statistically significant ( t=5.697, P< 0.001); the time taken to remove the dressing was (3.21 ± 1.24) min, (7.56 ± 1.88) min, and the difference was statistically significant ( t=11.917, P<0.001). Conclusions:The penile dorsal extension bandaging method after concealed penis surgery can reduce the number of calls to medical staff, reduce the time of dressing removal and the degree of pain in children.

Objective To evaluate the outcomes of penoplasty with penile frenulum lengthening for concealed penis in children.Methods From March 2014 to March 2016,a total of 233 patients with concealed penis who enderwent penoplasty with penile frenulum lengthening were enrolled.The everage age at surgery was 3.7 years (1 year and seven months-12 years).There were 73 cases with obvious small urinary stream,dysuria,or prepuce dilatation when urinating;41 cases with the history of recurrent infections of prepuces;and 9 cases with the history of urinary tract infections.During operation,incise the back side center of the outer plate of the prepuce and fully release the ring-type funicular tissue between the inner and outer plates of the prepuce to make the inner plate fully swell out.After the prepuce is upturned,cut the penile frenulum at the coronary sulcus.Make a V-shaped cut on the left and right sides of the far end of the outer plate cut of the prepuce from the cut of the penile frenulum.Lengthen penile frenulum after the prepuce is pressed off.Cover the dorsal side of the penis with the inner plate of the prepuce and cover its ventral side with its outer plate.Results Mean surgical time for patients was 38 min (30-55 min).All operations were completed successfully and the post-operation follow-up lasts 3 months to 2 years.For all cases,the appearance of the penis is improved.The penis stretches out,the balanus is exposed,the prepuce has no obvious swelling and the scrotum angle of the penis is obvious.No phimosis relapses and there is no obvious scar hyperplasia.Conclusions This surgical procedure is an effective treatment choice for concealed penis.It provides a good cosmetic result.