Objective:To explore the expression and mechanism of lncRNA MALAT1 in negative sputum for tuberculous bac-terium.Methods:Expression of lncRNA MALAT1 in peripheral blood mononuclear cells(PBMC)of patients with positive sputum bacteria(Positive group)and negative sputum bacteria(Negative group)and healthy volunteers(HC group)was detected by RT-PCR.ELISA was used to detect expression levels of TNF-α,IL-1β and IL-6 in plasma.Expression of lncRNA MALAT1 in mice macro-phages RAW264.7 was silenced by siRNA interference,and RAW264.7 cells were infected with mycobacterium tuberculosis(MTB)H37Rv.Cells were divided into four groups:Control group,Control+MTB group,MTB+si-NC group and MTB+si-MALAT1 group.Proliferation activity of RAW264.7 cells in each group was detected by CCK-8 method.The number of MTB in each group was detected by CFU.Expressions of TNF-α,IL-1β and IL-6 in supernatant of RAW264.7 cells were detected by ELISA.Results:Compared with HC group,expressions of lncRNA MALAT1 in PBMC,and TNF-α,IL-1β and IL-6 in plasma were significantly increased in Positive group and Negative group(P<0.01).Compared with Control group,expression level of lncRNA MALAT1,proliferation activity,CFU value,and concentrations of TNF-α,IL-1β and IL-6 in supernatant of Control+MTB group,MTB+si-NC group and MTB+si-MALAT1 group were significantly increased(P<0.05).Compared with MTB+si-NC group,the above detection indexes in MTB+si-MALAT1 group were significantly decreased(P<0.05),while there was no significant difference in Control+MTB group(P>0.05).Conclusion:The significantly increased expression of MALAT1 in patients with negative sputum for tuberculous bacterium is positively correlated with expression of plasma inflammatory factors,while the silence of MALAT1 expression can reduce MTB induced inflammatory response by inhibiting the proliferation and phagocytosis of MTB infected macrophages.

Objective:To explore the expression and mechanism of lncRNA MALAT1 in negative sputum for tuberculous bac-terium.Methods:Expression of lncRNA MALAT1 in peripheral blood mononuclear cells(PBMC)of patients with positive sputum bacteria(Positive group)and negative sputum bacteria(Negative group)and healthy volunteers(HC group)was detected by RT-PCR.ELISA was used to detect expression levels of TNF-α,IL-1β and IL-6 in plasma.Expression of lncRNA MALAT1 in mice macro-phages RAW264.7 was silenced by siRNA interference,and RAW264.7 cells were infected with mycobacterium tuberculosis(MTB)H37Rv.Cells were divided into four groups:Control group,Control+MTB group,MTB+si-NC group and MTB+si-MALAT1 group.Proliferation activity of RAW264.7 cells in each group was detected by CCK-8 method.The number of MTB in each group was detected by CFU.Expressions of TNF-α,IL-1β and IL-6 in supernatant of RAW264.7 cells were detected by ELISA.Results:Compared with HC group,expressions of lncRNA MALAT1 in PBMC,and TNF-α,IL-1β and IL-6 in plasma were significantly increased in Positive group and Negative group(P<0.01).Compared with Control group,expression level of lncRNA MALAT1,proliferation activity,CFU value,and concentrations of TNF-α,IL-1β and IL-6 in supernatant of Control+MTB group,MTB+si-NC group and MTB+si-MALAT1 group were significantly increased(P<0.05).Compared with MTB+si-NC group,the above detection indexes in MTB+si-MALAT1 group were significantly decreased(P<0.05),while there was no significant difference in Control+MTB group(P>0.05).Conclusion:The significantly increased expression of MALAT1 in patients with negative sputum for tuberculous bacterium is positively correlated with expression of plasma inflammatory factors,while the silence of MALAT1 expression can reduce MTB induced inflammatory response by inhibiting the proliferation and phagocytosis of MTB infected macrophages.

Objective:To assess the diagnostic efficiency of long-read sequencing (LRS) for the determination of CYP21A1P/ CYP21A2 and TNXA/ TNXB fusion genotypes among children with 21-hydroxylase deficiency (21-OHD) and explore their clinical characteristics. Methods:LRS sequencing was carried out on 30 children diagnosed with 21-OHD at the Department of Endocrinology, Fujian Children′s Hospital between November 2022 and September 2023 by clinical symptoms or conventional Sanger sequencing combined with multiple ligation-dependent probe amplification (MLPA). The results of the two methods were compared. Clinical data of the children were collected and analyzed. This study has been approved by the Medical Ethic Committee of the Fujian Children Hospital(Ethic No. 2022ETKLR10024).Results:Of the 30 children with 21-OHD, 11 (36.7%) were found to carry CYP21A1P/ CYP21A2 and TNXA/ TNXB fusion genes by LRS. The most common type of fused CYP21A1P/ CYP21A2 gene was CH-1 (61.5%), and 1 (3.3%) was found to harbor TNXA/ TNXB CH-1. 11 cases (36.7%) were found to carry large deletions by Sanger sequencing combined with MLPA, with the most common one being CYP21A2 exons 1-3 del (61.5%), which was followed by CYP21A2 exons 1-7 del (23.1%). Follow up of 11 patients carrying a fusion gene revealed that 6 were sale wasting (SW) types, 5 were simple virilizing (SV) types, whilst no non-classical (NC) type was found. Four girls had presented with central precocious puberty (CPP). One child carrying TNXA/ TNXB CH-1 had presented with CAH-X syndrome. Conclusion:Compared with Sanger sequencing combined with MLPA detection method, LRS sequencing was able to differentiate the subtypes of CYP21A1P/ CYP21A2 and TNXA/ TNXB fusion genes, pinpoint the breakpoints of the deletions, and directly determine the cis-trans position without the need to analyze the genotype of the pedigree members, which has provided a reliable method for the typing of 21-OHD. As some fusion genes may retain 21-hydroxylase activity, female carriers may have a higher incidence of CPP.

OBJECTIVE: To assess the diagnostic efficiency of long-read sequencing (LRS) for the determination of CYP21A1P/CYP21A2 and TNXA/TNXB fusion genotypes among children with 21-hydroxylase deficiency (21-OHD) and explore their clinical characteristics. METHODS: LRS sequencing was carried out on 30 children diagnosed with 21-OHD at the Department of Endocrinology, Fujian Children's Hospital between November 2022 and September 2023 by clinical symptoms or conventional Sanger sequencing combined with multiple ligation-dependent probe amplification (MLPA). The results of the two methods were compared. Clinical data of the children were collected and analyzed. This study has been approved by the Medical Ethics Committee of the Fujian Children's Hospital (Ethic No. 2022ETKLR10024). RESULTS: Of the 30 children with 21-OHD, 11 (36.7%) were found to carry CYP21A1P/CYP21A2 and TNXA/TNXB fusion genes by LRS. The most common type of fused CYP21A1P/CYP21A2 gene was CH-1 (72.7%), and 1 (3.3%) was found to harbor TNXA/TNXB CH-1. Eleven cases (36.7%) were found to carry large deletions by Sanger sequencing combined with MLPA, with the most common one being CYP21A2 exons 1-3 del (72.7%), which was followed by CYP21A2 exons 1-7 del (18.2%). Follow up of 11 patients carrying a fusion gene revealed that 6 were sale wasting (SW) types, 5 were simple virilizing (SV) types, whilst no non-classical (NC) type was found. Four girls had presented with central precocious puberty (CPP). One child carrying TNXA/TNXB CH-1 had presented with CAH-X syndrome. CONCLUSION Compared with Sanger sequencing combined with MLPA detection method, LRS sequencing was able to differentiate the subtypes of CYP21A1P/CYP21A2 and TNXA/TNXB fusion genes, pinpoint the breakpoints of the deletions, and directly determine the cis-trans position without the need to analyze the genotype of the pedigree members, which has provided a reliable method for the typing of 21-OHD. As some fusion genes may retain 21-hydroxylase activity, female carriers may have a higher incidence of CPP.
Humans ; Steroid 21-Hydroxylase/genetics* ; Adrenal Hyperplasia, Congenital/genetics* ; Child ; Female ; Male ; Child, Preschool ; Tenascin/genetics* ; Infant ; Genotype ; Sequence Analysis, DNA/methods* ; Pseudogenes

OBJECTIVE To study the extraction technology of Sophora flavescens-Phellodendron chinense drug pair and provide a reference for the development of new drugs for the treatment of anorectal diseases. METHODS Using the contents of total alkaloids of S. flavescens （matrine+oxymatrine）， berberine hydrochloride and total flavonoid， and extract yield as evaluation indicators， analytic hierarchy process-entropy weight method was used to calculate the weight coefficient of each indicator， and was combined with Box-Behnken design-response surface method to study the extraction technology of S. flavescens-P. chinense drug pair and verify it. RESULTS The optimal extraction technology of S. flavescens-P. chinense drug pair was immersed in 12-fold amount of 58% ethanol for 30 minutes and extracted twice， each time for 120 minutes. The relative error between the verification experimental results and the predicted value was 1.88%. CONCLUSIONS The obtained extraction technology is stable and feasible and can provide reference for the application of S. flavescens-P. chinense drug pair and development of new drugs.

Objective With the assistance of 3D visualization and real-time navigation technologies,the tumors in the parapharyngeal and lateral skull base should be removed through oral the approach with endoscopy.Methods The preoperative CT data of eight patients with parapharyngeal or lateral skull base soft tissue tumors were modeled,and the anatomical position relationship between the tumor and surrounding blood vessels and other important structures was re-constructed using 3D visualization technology,and preop-erative design was performed.The intraoperative oral ap-proach and real-time navigation guidance were adopted in the endoscopic resection of soft tissue tumors in the parapharyngeal and lateral skull base,and the clinical ap-plication value of this method was evaluated.Results The blood loss during the operation was controlled within 150 mL,and the average blood loss was approximately 125 mL.The incidence of postoperative complications was low,and patients could recover well through functional training.The oral approach did not leave any wounds nor scars on the patient's facial skin after the operation and had no effect on the patient's appearance.Conclusion The combination of 3D visualization technology,intraoperative real-time navigation,and endoscopy provides a beautiful,safe,and minimally invasive surgical method for patients with parapharyngeal or lateral skull base tumors.

Objective:To assess the diagnostic efficiency of long-read sequencing (LRS) for the determination of CYP21A1P/ CYP21A2 and TNXA/ TNXB fusion genotypes among children with 21-hydroxylase deficiency (21-OHD) and explore their clinical characteristics. Methods:LRS sequencing was carried out on 30 children diagnosed with 21-OHD at the Department of Endocrinology, Fujian Children′s Hospital between November 2022 and September 2023 by clinical symptoms or conventional Sanger sequencing combined with multiple ligation-dependent probe amplification (MLPA). The results of the two methods were compared. Clinical data of the children were collected and analyzed. This study has been approved by the Medical Ethic Committee of the Fujian Children Hospital(Ethic No. 2022ETKLR10024).Results:Of the 30 children with 21-OHD, 11 (36.7%) were found to carry CYP21A1P/ CYP21A2 and TNXA/ TNXB fusion genes by LRS. The most common type of fused CYP21A1P/ CYP21A2 gene was CH-1 (61.5%), and 1 (3.3%) was found to harbor TNXA/ TNXB CH-1. 11 cases (36.7%) were found to carry large deletions by Sanger sequencing combined with MLPA, with the most common one being CYP21A2 exons 1-3 del (61.5%), which was followed by CYP21A2 exons 1-7 del (23.1%). Follow up of 11 patients carrying a fusion gene revealed that 6 were sale wasting (SW) types, 5 were simple virilizing (SV) types, whilst no non-classical (NC) type was found. Four girls had presented with central precocious puberty (CPP). One child carrying TNXA/ TNXB CH-1 had presented with CAH-X syndrome. Conclusion:Compared with Sanger sequencing combined with MLPA detection method, LRS sequencing was able to differentiate the subtypes of CYP21A1P/ CYP21A2 and TNXA/ TNXB fusion genes, pinpoint the breakpoints of the deletions, and directly determine the cis-trans position without the need to analyze the genotype of the pedigree members, which has provided a reliable method for the typing of 21-OHD. As some fusion genes may retain 21-hydroxylase activity, female carriers may have a higher incidence of CPP.

Objective:To study the clinical effect of dynamic reconstruction of lower lip defect after squamous cell cancer resection by using unilateral Gillies fan flap and perioral muscle restoration.Methods:From Feburary 2017 to January 2020, 8 patients (6 female and 2 male, aged 55-81, 62.9 in average)diagnosed lower lip squamous cell carcinoma were accepted treatment in the Department of Otolaryngology Head & Neck Surgery, the First Affiliated Hospital of Xiamen University. All the moderate lower lip defect after cancer resection were repaired with unilateral Gillies fan flap with perioral muscle restoration under general anesthesia. The length of the vermilion of the upper lip in the Gillies fan flap was designed to be 1/3 of the length of the vermilion of the lower lip defect. The composite axial flap was prepared with the blood supply from contralateral upper lip artery-ipsilateral upper lip artery-mouth corner part of facial artery-ipsilateral lower lip artery.The end of all the perioral muscles were also prepared. After reconstruction of the continuity of the orbicularis oris, the ends of these perioral radiation muscles were restored in the position where they should be in the reconstructed lip.These perioral muscles training were undergone for 6 months after operation. After the reconstructed lip tend to be stable, measurement and calculation were carried out to evaluate the appearance, opening and closing functions of the mouth.Results:In our series of 8 cases, the length of the vermilion of the upper/lower lip was 52.4 mm(46.0-60.0 mm) in average; the circumference of the vermilion was 104.8 mm(92-120 mm)in average; the length of the defect of lower lip was 27.6 mm (21.0-35.0 mm)in average, 52.5% (42.0-67.3%) of the vermilion of lower lip. Six cases were using the right-side flap, and the other 2 cases were left. The length of the vermilion of the upper lip in the Gillies fan flap was 9.5 mm(7.0-12.0 mm)in average.All cases healed well with inconspicuous scar. No infection, hematoma or other surgical complications.No local or distant recurrence of cancer was observed during the follow-up period. Numbness of the lower lip happened posto-peration, and gradually disappeared 6 months later.Evaluation was conducted according to the measurement result preoperatively and 6 months postoperatively. (1) Appearance: the reconstructed lips were naturally symmetrical. Both side of the mouth corner were in the same horizontal level. The upper lip was lengthened with average increase of 6.8 mm and average increase rate of 15.9%. The lower lip was lengthened with average increase of 15.4 mm and average increase rate of 45.3%. In the rest situation, the circumference of the lips decreased by 5.5 mm in average compared with that preoperatively, which was 94.9% of the lip preoperatively, basically rehabilitate to the shape preoperatively.(2) Mouth-opening function: 1 week postoperation, the opening shape of mouth in all the cases was skewed with an about 1.5 fingers mouth-opening. Six months later, the opening shape of mouth was basically symmetrical round with a 3 fingers mouth-opening; in maximum mouth opening situation, the circumference of the lips decreased by 46.5 mm in average compared with that preoperatively, which was 73.1% of the lip in maximum mouth opening situation preoperatively, basically meeting physiological needs. (3) Mouth-closing function: All patient shave complete mouth-closing function and speech function.No leakage when making closed lip sound and whistling. All patients were satisfied with the oral appearance and the function of opening and closing mouth.Conclusions:Unilateral Gillies fan flap with perioral muscle restoration is a reliable method to repair the moderate full-thickness defect of the lower lip after cancer resection. After dynamic reconstruction of the perioral muscle group, the insufficient tissue of lips can be extended to obtain sufficient mouth opening, natural corner of the mouth and symmetrical lip appearance.

Objective:To study the clinical effect of dynamic reconstruction of lower lip defect after squamous cell cancer resection by using unilateral Gillies fan flap and perioral muscle restoration.Methods:From Feburary 2017 to January 2020, 8 patients (6 female and 2 male, aged 55-81, 62.9 in average)diagnosed lower lip squamous cell carcinoma were accepted treatment in the Department of Otolaryngology Head & Neck Surgery, the First Affiliated Hospital of Xiamen University. All the moderate lower lip defect after cancer resection were repaired with unilateral Gillies fan flap with perioral muscle restoration under general anesthesia. The length of the vermilion of the upper lip in the Gillies fan flap was designed to be 1/3 of the length of the vermilion of the lower lip defect. The composite axial flap was prepared with the blood supply from contralateral upper lip artery-ipsilateral upper lip artery-mouth corner part of facial artery-ipsilateral lower lip artery.The end of all the perioral muscles were also prepared. After reconstruction of the continuity of the orbicularis oris, the ends of these perioral radiation muscles were restored in the position where they should be in the reconstructed lip.These perioral muscles training were undergone for 6 months after operation. After the reconstructed lip tend to be stable, measurement and calculation were carried out to evaluate the appearance, opening and closing functions of the mouth.Results:In our series of 8 cases, the length of the vermilion of the upper/lower lip was 52.4 mm(46.0-60.0 mm) in average; the circumference of the vermilion was 104.8 mm(92-120 mm)in average; the length of the defect of lower lip was 27.6 mm (21.0-35.0 mm)in average, 52.5% (42.0-67.3%) of the vermilion of lower lip. Six cases were using the right-side flap, and the other 2 cases were left. The length of the vermilion of the upper lip in the Gillies fan flap was 9.5 mm(7.0-12.0 mm)in average.All cases healed well with inconspicuous scar. No infection, hematoma or other surgical complications.No local or distant recurrence of cancer was observed during the follow-up period. Numbness of the lower lip happened posto-peration, and gradually disappeared 6 months later.Evaluation was conducted according to the measurement result preoperatively and 6 months postoperatively. (1) Appearance: the reconstructed lips were naturally symmetrical. Both side of the mouth corner were in the same horizontal level. The upper lip was lengthened with average increase of 6.8 mm and average increase rate of 15.9%. The lower lip was lengthened with average increase of 15.4 mm and average increase rate of 45.3%. In the rest situation, the circumference of the lips decreased by 5.5 mm in average compared with that preoperatively, which was 94.9% of the lip preoperatively, basically rehabilitate to the shape preoperatively.(2) Mouth-opening function: 1 week postoperation, the opening shape of mouth in all the cases was skewed with an about 1.5 fingers mouth-opening. Six months later, the opening shape of mouth was basically symmetrical round with a 3 fingers mouth-opening; in maximum mouth opening situation, the circumference of the lips decreased by 46.5 mm in average compared with that preoperatively, which was 73.1% of the lip in maximum mouth opening situation preoperatively, basically meeting physiological needs. (3) Mouth-closing function: All patient shave complete mouth-closing function and speech function.No leakage when making closed lip sound and whistling. All patients were satisfied with the oral appearance and the function of opening and closing mouth.Conclusions:Unilateral Gillies fan flap with perioral muscle restoration is a reliable method to repair the moderate full-thickness defect of the lower lip after cancer resection. After dynamic reconstruction of the perioral muscle group, the insufficient tissue of lips can be extended to obtain sufficient mouth opening, natural corner of the mouth and symmetrical lip appearance.

Objective:To study the clinical effect of dynamic reconstruction of lower lip defect after squamous cell cancer resection by using unilateral Gillies fan flap and perioral muscle restoration.Methods:From Feburary 2017 to January 2020, 8 patients (6 female and 2 male, aged 55-81, 62.9 in average)diagnosed lower lip squamous cell carcinoma were accepted treatment in the Department of Otolaryngology Head & Neck Surgery, the First Affiliated Hospital of Xiamen University. All the moderate lower lip defect after cancer resection were repaired with unilateral Gillies fan flap with perioral muscle restoration under general anesthesia. The length of the vermilion of the upper lip in the Gillies fan flap was designed to be 1/3 of the length of the vermilion of the lower lip defect. The composite axial flap was prepared with the blood supply from contralateral upper lip artery-ipsilateral upper lip artery-mouth corner part of facial artery-ipsilateral lower lip artery.The end of all the perioral muscles were also prepared. After reconstruction of the continuity of the orbicularis oris, the ends of these perioral radiation muscles were restored in the position where they should be in the reconstructed lip.These perioral muscles training were undergone for 6 months after operation. After the reconstructed lip tend to be stable, measurement and calculation were carried out to evaluate the appearance, opening and closing functions of the mouth.Results:In our series of 8 cases, the length of the vermilion of the upper/lower lip was 52.4 mm(46.0-60.0 mm) in average; the circumference of the vermilion was 104.8 mm(92-120 mm)in average; the length of the defect of lower lip was 27.6 mm (21.0-35.0 mm)in average, 52.5% (42.0-67.3%) of the vermilion of lower lip. Six cases were using the right-side flap, and the other 2 cases were left. The length of the vermilion of the upper lip in the Gillies fan flap was 9.5 mm(7.0-12.0 mm)in average.All cases healed well with inconspicuous scar. No infection, hematoma or other surgical complications.No local or distant recurrence of cancer was observed during the follow-up period. Numbness of the lower lip happened posto-peration, and gradually disappeared 6 months later.Evaluation was conducted according to the measurement result preoperatively and 6 months postoperatively. (1) Appearance: the reconstructed lips were naturally symmetrical. Both side of the mouth corner were in the same horizontal level. The upper lip was lengthened with average increase of 6.8 mm and average increase rate of 15.9%. The lower lip was lengthened with average increase of 15.4 mm and average increase rate of 45.3%. In the rest situation, the circumference of the lips decreased by 5.5 mm in average compared with that preoperatively, which was 94.9% of the lip preoperatively, basically rehabilitate to the shape preoperatively.(2) Mouth-opening function: 1 week postoperation, the opening shape of mouth in all the cases was skewed with an about 1.5 fingers mouth-opening. Six months later, the opening shape of mouth was basically symmetrical round with a 3 fingers mouth-opening; in maximum mouth opening situation, the circumference of the lips decreased by 46.5 mm in average compared with that preoperatively, which was 73.1% of the lip in maximum mouth opening situation preoperatively, basically meeting physiological needs. (3) Mouth-closing function: All patient shave complete mouth-closing function and speech function.No leakage when making closed lip sound and whistling. All patients were satisfied with the oral appearance and the function of opening and closing mouth.Conclusions:Unilateral Gillies fan flap with perioral muscle restoration is a reliable method to repair the moderate full-thickness defect of the lower lip after cancer resection. After dynamic reconstruction of the perioral muscle group, the insufficient tissue of lips can be extended to obtain sufficient mouth opening, natural corner of the mouth and symmetrical lip appearance.