Pyran- and furanocoumarins are key representatives of tetrahydropyrans and tetrahydrofurans, respectively, exhibiting diverse physiological and medical bioactivities. However, the biosynthetic mechanisms for their core structures remain poorly understood. Here we combined multiomics analyses of biosynthetic enzymes in Peucedanum praeruptorum and in vitro functional verification and identified two types of key enzymes critical for pyran and furan ring biosynthesis in plants. These included three distinct P. praeruptorum prenyltransferases (PpPT1-3) responsible for the prenylation of the simple coumarin skeleton 7 into linear or angular precursors, and two novel CYP450 cyclases (PpDC and PpOC) crucial for the cyclization of the linear/angular precursors into either tetrahydropyran or tetrahydrofuran scaffolds. Biochemical analyses of cyclases indicated that acid/base-assisted epoxide ring opening contributed to the enzyme-catalyzed tetrahydropyran and tetrahydrofuran ring refactoring. The possible acid/base-assisted catalytic mechanisms of the identified cyclases were theoretically investigated and assessed using site-specific mutagenesis. We identified two possible acidic amino acids Glu303 in PpDC and Asp301 in PpOC as vital in the catalytic process. This study provides new enzymatic tools in the epoxide formation/epoxide-opening mediated cascade reaction and exemplifies how plants become chemically diverse in terms of enzyme function and catalytic process.

Objective:To investigate the application and effect of frontal-orbital reconstruction and distraction osteogenesis in the treatment of Crouzon syndrome using three-dimensiond reconstruction mold (3D-RM).Methods:Retrospective analysis was performed on the clinical data of children with Crouzon syndrome who were treated in the Department of Burning and Plastic Surgery, Children’s Hospital of Nanjing Medical University from January 2018 to December 2021. These children were divided into two groups. The study group performed fronto-orbital reconstruction combined with distraction osteogenesis using 3D-RM, while the control group performed the surgery without using 3D-RM. Intraoperative blood loss, operation time, duration of hospital stay, complication rate, cranial index, frontal and orbital retraction rate and postoperative frontal and orbital contour were compared between the two groups. The independent sample t-test was used to compare the differences in intraoperative blood loss, operation duration, hospital stay, postoperative cranial index and fronto-orbital retraction rate between the two groups. The Chi-square test was used to compare the differences in the incidence of postoperative complications between the two groups. Results:The study group included 10 children, including 6 males and 4 females, ranging in age from 9 to 37 months, with an average age of 19 months. In the control group, 8 children were included, including 5 males and 3 females, ranging in age from 8 to 46 months, with an average age of 25 months. The study group was followed up for 12 to 36 months, with an average of 18 months, and the head shape was better improved. The control group was followed up for 12 to 30 months, with an average of 18 months. No serious complications occurred and the head deformity was improved. There were no significant differences in intraoperative blood loss, operation duration, hospital stay, cranial index, fronto-orbital retraction rate and complication rate between the two groups (all P>0.05). The forehead shape of the study group was more symmetrical than that of the control group, the difference was statistically significant ( P<0.05). Conclusion:The effect of frontal orbital reconstruction and distraction osteogenesis using 3D-RM in the treatment of Crouzon syndrome is positive, the head shape and orbit are significantly improved. There were no serious complications after operation. It can play a good auxiliary role in the operation.

Objective:To investigate the application and effect of frontal-orbital reconstruction and distraction osteogenesis in the treatment of Crouzon syndrome using three-dimensiond reconstruction mold (3D-RM).Methods:Retrospective analysis was performed on the clinical data of children with Crouzon syndrome who were treated in the Department of Burning and Plastic Surgery, Children’s Hospital of Nanjing Medical University from January 2018 to December 2021. These children were divided into two groups. The study group performed fronto-orbital reconstruction combined with distraction osteogenesis using 3D-RM, while the control group performed the surgery without using 3D-RM. Intraoperative blood loss, operation time, duration of hospital stay, complication rate, cranial index, frontal and orbital retraction rate and postoperative frontal and orbital contour were compared between the two groups. The independent sample t-test was used to compare the differences in intraoperative blood loss, operation duration, hospital stay, postoperative cranial index and fronto-orbital retraction rate between the two groups. The Chi-square test was used to compare the differences in the incidence of postoperative complications between the two groups. Results:The study group included 10 children, including 6 males and 4 females, ranging in age from 9 to 37 months, with an average age of 19 months. In the control group, 8 children were included, including 5 males and 3 females, ranging in age from 8 to 46 months, with an average age of 25 months. The study group was followed up for 12 to 36 months, with an average of 18 months, and the head shape was better improved. The control group was followed up for 12 to 30 months, with an average of 18 months. No serious complications occurred and the head deformity was improved. There were no significant differences in intraoperative blood loss, operation duration, hospital stay, cranial index, fronto-orbital retraction rate and complication rate between the two groups (all P>0.05). The forehead shape of the study group was more symmetrical than that of the control group, the difference was statistically significant ( P<0.05). Conclusion:The effect of frontal orbital reconstruction and distraction osteogenesis using 3D-RM in the treatment of Crouzon syndrome is positive, the head shape and orbit are significantly improved. There were no serious complications after operation. It can play a good auxiliary role in the operation.

Objective: To investigate the epidemiological characteristics of malaria in Hangzhou City from 2004 to 2021, so as to provide the evidence for formulating the post-elimination control strategy for malaria in Hangzhou City. Methods: The epidemic situation of malaria in Hangzhou City from 2004 to 2021 were collected from the National Information System for Disease Control and Prevention in China, and the temporal, spatial and human distributions of malaria cases and the source of malaria infections were analyzed in Hangzhou City during the pre-elimination stage (2004 to 2009), the elimination stage (2010 to 2015) and the post-elimination stage (2016 to 2021). Results: Totally 602 malaria cases were reported in Hangzhou City from 2004 to 2021,and the annual mean incidence of malaria was 0.22/105, 0.20/105 and 0.18/105 during the pre-elimination, elimination and post-elimination stages, appearing a tendency towards a decline. Men accounted for 63.96%, 85.07% and 93.75% of all malaria cases and there were 67.86%, 82.84% and 80.00% of cases at ages of 18 to 50 years during the pre-elimination, elimination and post-elimination stages, both appearing a tendency towards a decline (χ2trend=56.748, P<0.001; χ2trend=39.971, P<0.001). The predominant occupation of malaria cases shifted from farmers or migrant workers to multiple occupations, and the proportion of commercial servants increased from 4.87% during the pre-elimination stage to 24.38% during the post-elimination stage (χ2trend=73.308, P<0.001). The proportion of Plasmodium vivax malaria cases reduced from 96.43% during the pre-elimination stage to 7.50% during the post-elimination stage, and the proportion of P. falciparum malaria cases increased from 3.57% to 71.25%, while P. ovale, P. malariae and mixed infections were identified since 2010. There was a significant season-specific incidence of P. vivax malaria during the pre-elimination stage, and the period between May and October was an epidemic season; however, there was no season-specific incidence of P. vivax malaria during the elimination and post-elimination stages. The regional distribution of malaria cases presented a tendency towards a shift from suburb and rural areas to urban areas (χ2trend=74.229, P<0.001). No local cases were detected in Hangzhou City since 2010, and 94.22% of malaria cases were overseas imported cases after malaria elimination, including 90.61% from Africa. Conclusions oung and middle-aged men were high-risk populations for malaria in Hangzhou City from 2004 to 2021, and overseas commercial servants gradually became the predominant source of malaria infections, with malaria parasite species tending to be diverse. Improving the management of overseas imported cases and timely identification and treatment of cases are major interventions to consolidate malaria elimination achievements in Hangzhou City.

Objective:To investigate the clinical effect of mandible distraction combined with concentrated growth factor (CGF) for the treatment of Pierre Robin sequence (PRS).Methods:This retrospective research recruited clinical data of PRS neonates from January 2018 to December 2019 in the Department of Burns and Plastic Surgery, Children’s Hospital of Nanjing Medical University. All cases were divided into CGF group (osteotomy gap filled with CGF gel during distraction osteotomy), and NO-CGF group (no CGF gel used during distraction osteotomy). Bilateral internal distractors, which could be extended by 2 cm, were fixed on mandible after mandible osteotomy. Immediate distraction for 5 mm was performed in all children during the operation. After a latency period of 5 days, the jaw was extended 0.8 to 1.2 mm once a day, and the extension period was 10 to 15 days. The distractors were surgically removed after 3 months of application. Both postoperative extubation time and complications were analyzed.Results:CGF group: the male∶female ratio was 32∶13, with age ranged 3 to 28 days and weight ranged 2.5 to 3.2 kg. NO-CGF group: the male∶female ratio was 40∶15, with age ranged 3 to 28 days and weight ranged 2.6 to 3.3 kg. The total mandibular distraction length was 15 to 20 mm, and no serious complications (such as death or osteonecrosis) were observed during 3 to 6 months postoperative follow-up. 68 cases (30 in CGF group and 38 in NO-CGF group) experienced immediate extubation after surgery, and 32 cases (15 in CGF group and 17 in NO-CGF group) were extubated 4 to 6 days after the operation. For postoperative complications, CGF group (1 case with infection) and NO-CGF group (4 cases with infection, 2 cases of screw loss, 8 cases of delayed bone union, and 4 cases of mandibular retraction) were compared respectively. Statistical significance in the complications of delayed bone union was found between two groups ( P<0.05). Conclusions:Mandible distraction combined with CGF is a promising technique for the management of PRS as evidenced by the low incidence of complications and improved bone union.

Objective:To learn about the knowledge and practice of iodine deficiency disorders (IDD) prevention and control among main caregivers of pregnant women in trimester and its related influencing factors, so as to provide a scientific basis for effectively improving the iodine nutrition level of pregnant women.Methods:In April 2020, using the stratified sampling method, 2 districts and counties were selected in the urban, suburban and rural areas of Hangzhou City, respectively, and 5 maternal and child health care institutions were selected as the survey sites in each district and county according to the five directions of East, West, South, North and Middle. A face-to-face questionnaire investigation was used to investigate the knowledge and practice of IDD prevention and control among main caregivers of pregnant women.Results:Totally 601 caregivers of pregnant women were investigated. Among them, 68.9% (414/601) knew that eating iodized salt was the best way to prevent IDD. However, 15.0% (90/601) thought that eating seafood was the best way to prevent IDD. And 41.9% (252/601) caregivers of pregnant women would actively purchase iodine-rich food for cooking, and 63.6% (382/601) would choose iodized salt. There were statistically significant differences in the knowledge and practice of IDD prevention and control among caregivers of pregnant women in urban, suburban and rural areas ( P < 0.05). And 50.3% (87/173) male caregivers would actively purchase iodine-rich food for cooking, which was higher than that of female caregivers [38.6% (165/428)], the difference was statistically significant (χ 2 = 6.97, P = 0.008). There was a statistically significant difference between the caregivers of pregnant women of different ages in actively purchasing iodine-rich food for cooking (χ 2 = 12.61, P = 0.013). And there were statistically significant differences between the caregivers of pregnant women with different educational levels in actively purchasing iodine-rich food for cooking and choosing iodized salt (χ 2 = 29.34, 39.42, P < 0.001). Conclusions:The awareness of knowledge about IDD prevention and control and the formation of prevention and control behavior among caregivers of pregnant women in Hangzhou City are poor. Active education of caregivers of pregnant women should be strengthened to promote maternal and infant health.

Objective:To investigate the clinical effect of mandible distraction combined with concentrated growth factor (CGF) for the treatment of Pierre Robin sequence (PRS).Methods:This retrospective research recruited clinical data of PRS neonates from January 2018 to December 2019 in the Department of Burns and Plastic Surgery, Children’s Hospital of Nanjing Medical University. All cases were divided into CGF group (osteotomy gap filled with CGF gel during distraction osteotomy), and NO-CGF group (no CGF gel used during distraction osteotomy). Bilateral internal distractors, which could be extended by 2 cm, were fixed on mandible after mandible osteotomy. Immediate distraction for 5 mm was performed in all children during the operation. After a latency period of 5 days, the jaw was extended 0.8 to 1.2 mm once a day, and the extension period was 10 to 15 days. The distractors were surgically removed after 3 months of application. Both postoperative extubation time and complications were analyzed.Results:CGF group: the male∶female ratio was 32∶13, with age ranged 3 to 28 days and weight ranged 2.5 to 3.2 kg. NO-CGF group: the male∶female ratio was 40∶15, with age ranged 3 to 28 days and weight ranged 2.6 to 3.3 kg. The total mandibular distraction length was 15 to 20 mm, and no serious complications (such as death or osteonecrosis) were observed during 3 to 6 months postoperative follow-up. 68 cases (30 in CGF group and 38 in NO-CGF group) experienced immediate extubation after surgery, and 32 cases (15 in CGF group and 17 in NO-CGF group) were extubated 4 to 6 days after the operation. For postoperative complications, CGF group (1 case with infection) and NO-CGF group (4 cases with infection, 2 cases of screw loss, 8 cases of delayed bone union, and 4 cases of mandibular retraction) were compared respectively. Statistical significance in the complications of delayed bone union was found between two groups ( P<0.05). Conclusions:Mandible distraction combined with CGF is a promising technique for the management of PRS as evidenced by the low incidence of complications and improved bone union.

Objective:To investigate the safety and therapeutic effect of surgical resection combined with radiofrequency ablation on facial plexiform neurofibroma.Methods:The data of children with neurofibromatosis type Ⅰ treated by surgical excision combined with radiofrequency ablation in the Department of Burn and Plastic Surgery of Children’s Hospital of Nanjing Medical University from April 1, 2016 to April 1, 2020 was retrospectively analyzed. All patients were treated with partial tumor resection and radiofrequency ablation under general anesthesia. During the operation, the tumor body is gradually exposed through the hairline concealed incision. The tumor in the field of vision was removed to the maximum extent. Radiofrequency ablation was performed on distant tumor parts that was visible but inoperable, so as to achieve no residual tumor. Radiofrequency ablation was abandoned if intraoperative exploration revealed tumor invasion of facial nerve, deep fascia and orbit. The maximum output power of the radiofrequency ablation machine used during the operation was 200 W, and the exposed length of the electrode was 3 cm. The radio frequency machine could automatically adjust the power during the ablation process according to the impedance of the tissue to output the optimal energy, and monitor the temperature of the surrounding tissue in real time during the ablation process. The distal tumor body was located by B-ultrasound, penetrated, and ablated by radiofrequency needle. The amount of intraoperative bleeding was estimated, and the postoperative drainage volume, drainage tube placement time and complications were observed and analyzed. The facial nerve injury was evaluated at the time of discharge and the first month after operation. Six months after operation, the scar was evaluated with the patient scar assessment scale (PSAS) in the patient and observer scar assessment scale (POSAS). The measurement data of normal distribution was expressed as Mean±SD.Results:A total of 5 children were included, including 3 males and 2 females, aged from 1 year and 2 months to 6 years and 10 months. The tumors were all located in the facial area. The range of invasion was superficial muscle layer, and the postoperative pathology indicated plexiform neurofibroma. The incision length of 5 patients was (5.4±0.4) cm, the intraoperative bleeding volume was (36.0±4.9) ml, the postoperative drainage volume was (4.2±0.7) ml, and the drainage tube was placed for (2.4±0.5) days. No local infection or skin ulceration occurred after operation, and the body temperature did not fluctuate significantly. Follow-up time ranged from 12 months to 18 months, the median follow-up time was 12 months. At the time of discharge and one month after operation, there was no difficulty in eyebrow lifting in all the patients. In the early postoperative period, the swelling of the operation site was obvious, and the swelling gradually decreased after 40 days at the latest. One child developed local skin cyanosis, which gradually subsided one month after the operation after cold compress. It was considered that it was caused by local subcutaneous slight bleeding. The local scar after operation was slight and concealed, and the PSAS score of the children 6 months after operation was 27.2±4.4. During the follow-up period, the original tumor body was stable, and no progressive enlargement of tumor body was found.Conclusions:Surgical resection combined with radiofrequency ablation for facial plexiform neurofibroma is safe and effective. Compared with traditional surgery, it can reduce local injury and facial changes, and can be used as an auxiliary treatment, but it cannot completely replace the traditional surgery.

Objective:To investigate the effect of intrafrontal-orbital distraction osteogenesis in the treatment for school-aged children with severe proptosis in Crouzon syndrome.Methods:We retrospectively analyzed the clinical data of school-aged children with Crouzon syndrome treated with intrafrontal orbital distraction osteotomy at the Children’s Hospital of Nanjing Medical University from January 2016 to February 2021, and counted and analyzed the clinical results, complications (cerebrospinal fluid leakage, infection, etc.), and surgical safety.Results:A total of 12 children, 9 males and 3 females, aged 6-12.1 years, with an average age of 7.1 years, were included; 4 were cranial, 6 were craniofacial, and 2 were other types. All were shown with obvious proptosis. The surgical procedure was successful. Follow-up ranged from 3 to 20 months, with a mean of 14 months. The postoperative distraction distance was (19.1±3.3) mm, ranging from 15.2 to 25.6 mm. And the mean cranial index (CI) before and after surgery were 97.5% and 80.0%, respectively, with good craniofacial appearance and better improvement of proptosis and posterior cranial flattening. There were no other complications but for one child with infection at the external site of the distraction rod.Conclusions:The effect of frontal-orbital distraction osteotomy in the treatment of Crouzon syndrome with severe proptosis in school-aged children was positive, with significant improvement in proptosis and low incidence of postoperative cerebrospinal fluid leakage and infection. This procedure is a good choice for older children with severe proptosis.

Objective:To investigate the safety and therapeutic effect of surgical resection combined with radiofrequency ablation on facial plexiform neurofibroma.Methods:The data of children with neurofibromatosis type Ⅰ treated by surgical excision combined with radiofrequency ablation in the Department of Burn and Plastic Surgery of Children’s Hospital of Nanjing Medical University from April 1, 2016 to April 1, 2020 was retrospectively analyzed. All patients were treated with partial tumor resection and radiofrequency ablation under general anesthesia. During the operation, the tumor body is gradually exposed through the hairline concealed incision. The tumor in the field of vision was removed to the maximum extent. Radiofrequency ablation was performed on distant tumor parts that was visible but inoperable, so as to achieve no residual tumor. Radiofrequency ablation was abandoned if intraoperative exploration revealed tumor invasion of facial nerve, deep fascia and orbit. The maximum output power of the radiofrequency ablation machine used during the operation was 200 W, and the exposed length of the electrode was 3 cm. The radio frequency machine could automatically adjust the power during the ablation process according to the impedance of the tissue to output the optimal energy, and monitor the temperature of the surrounding tissue in real time during the ablation process. The distal tumor body was located by B-ultrasound, penetrated, and ablated by radiofrequency needle. The amount of intraoperative bleeding was estimated, and the postoperative drainage volume, drainage tube placement time and complications were observed and analyzed. The facial nerve injury was evaluated at the time of discharge and the first month after operation. Six months after operation, the scar was evaluated with the patient scar assessment scale (PSAS) in the patient and observer scar assessment scale (POSAS). The measurement data of normal distribution was expressed as Mean±SD.Results:A total of 5 children were included, including 3 males and 2 females, aged from 1 year and 2 months to 6 years and 10 months. The tumors were all located in the facial area. The range of invasion was superficial muscle layer, and the postoperative pathology indicated plexiform neurofibroma. The incision length of 5 patients was (5.4±0.4) cm, the intraoperative bleeding volume was (36.0±4.9) ml, the postoperative drainage volume was (4.2±0.7) ml, and the drainage tube was placed for (2.4±0.5) days. No local infection or skin ulceration occurred after operation, and the body temperature did not fluctuate significantly. Follow-up time ranged from 12 months to 18 months, the median follow-up time was 12 months. At the time of discharge and one month after operation, there was no difficulty in eyebrow lifting in all the patients. In the early postoperative period, the swelling of the operation site was obvious, and the swelling gradually decreased after 40 days at the latest. One child developed local skin cyanosis, which gradually subsided one month after the operation after cold compress. It was considered that it was caused by local subcutaneous slight bleeding. The local scar after operation was slight and concealed, and the PSAS score of the children 6 months after operation was 27.2±4.4. During the follow-up period, the original tumor body was stable, and no progressive enlargement of tumor body was found.Conclusions:Surgical resection combined with radiofrequency ablation for facial plexiform neurofibroma is safe and effective. Compared with traditional surgery, it can reduce local injury and facial changes, and can be used as an auxiliary treatment, but it cannot completely replace the traditional surgery.