ObjectiveTo induce the rat model of ulcerative colitis （UC） with spleen-kidney Yang deficiency and liver depression， and explore the efficacy and mechanism of Sishenwan combined with Tongxie Yaofang （SSW&TXYF） based on the therapeutic principles of tonifying spleen， soothing liver， warming kidney， and astringing intestine. MethodSixty male SD rats were randomized into normal， model， mesalazine， and high-， medium-， and low-dose SSW&TXYF groups. The rats in other groups except the normal group were administrated with Sennae Folium decoction and hydrocortisone and received tail clamping for 14 days. On day 14， rats received enema with TNBS-ethanol solution to induce UC. The rats were administrated with corresponding drugs from day 15 of modeling， and the body weight and mental state were observed and recorded. The sucrose preference test was performed from day 25. On day 28， the rectal temperature was measured， and the rats were administrated with 3% D-xylose solution at a dose of 10 mL·kg^-1 by gavage. Blood was sampled 1 h later， from which the serum was collected for measurement of the D-xylose content. The serum， hippocampus， and colorectum samples of rats were collected on day 29. The levels of gastrin （GAS）， adrenocorticotropic hormone （ACTH）， corticosterone （CORT）， cyclic adenosine monophosphate （cAMP）， cyclic guanosine monophosphate （cGMP）， interleukin （IL）-4， tumor necrosis factor （TNF）-α， and interferon （IFN）-γ in the serum and 5-hydroxytryptamine （5-HT） in the hippocampus were determined by enzyme-linked immunosorbent assay. Hematoxylin-eosin staining was employed to reveal the colonic lesions. The mRNA and protein levels of p38 mitogen-activated protein kinase （MAPK）， extracellular signal-regulated kinase （ERK）， and c-Jun N-terminal kinase （JNK） in the colon tissue were determined by Real-time PCR and Western blot， respectively. ResultCompared with the normal group， the model group showed decreased body weight， anal temperature， and D-xylose content in the serum and increased GAS content （P<0.01）. The modeling led to cAMP/cGMP unbalance and decreased the ACTH and CORT content in the serum （P<0.01）， the preference for sucrose water， and the 5-HT content in the hippocampus （P<0.01）. Moreover， it shortened the colorectal length and caused massive infiltration of inflammatory cells and severe structural damage in the colon tissue. High， medium， and low doses of SSW&TXYF improved above indicators （P<0.05， P<0.01）， reduced inflammatory infiltration， and repaired the pathological damage of the tissue. Compared with the normal group， the model group showed lowered IL-4 level （P<0.01） and elevated TNF-α and IFN-γ levels （P<0.05， P<0.01） in the serum， as well as up-regulated expression of p38 MAPK， ERK， and JNK （P<0.05， P<0.01）. Compared with the model group， SSW&TXYF elevated the IL-4 level （P<0.01）， lowered the TNF-α and IFN-γ levels （P<0.05， P<0.01）， and down-regulated the mRNA and protein levels of p38 MAPK， ERK， and JNK （P<0.05， P<0.01）. ConclusionA rat model of UC with spleen-kidney Yang deficiency and liver depression was successfully established. SSW&TXYF can significantly mitigate this syndrome by reducing the inflammatory response in the colon and inhibiting the MAPK pathway.

Objective To investigate the impact of SARS-CoV-2 infection on male semen quality through meta-analysis and retrospec-tive study.Methods Literature retrieval was conducted in PubMed,CNKI,Wanfang Database and CBM database.Meta-analysis was performed using Stata 15.0.The male patients meeting the inclusion criteria from our hospital were enrolled as study subjects.General demographic data and semen parameters were collected.Single factor analysis of variance and graphing of semen parameters were con-ducted using GraphPad Prism 9.5.1.The test level was set at 0.05.Results A total of 9 studies involving 267 patients were included in the meta-analysis.There were no significant differences in sperm concentration and survival rate before and after SARS-CoV-2 infec-tion(P＞0.05).Semen volume and percentage of normal morphology significantly increased during 1-3 months after infection(P=0.005,P=0.010),with semen volume recovering to pre-infection level＞3 months later(P＞0.05).Sperm motility and progressive mo-tility increased＞3 months after infection(P=0.046,P=0.045),recovering to pre-infection levels(P=0.099,P=0.098).Sperm DNA fragmentation index may be temporarily elevated within 3 months after infection but gradually decreased＞3 months later.In the retrospective study of 8 cases,there were no statistical differences in semen parameters at different stages compared with pre-infection(P＞0.05),but semen parameters showed a negative trend during＜1 month after infection and recovered to pre-infection levels＞3 months later.Conclusion The results of this laboratory study are basically consistent with the meta-analysis.SARS-CoV-2 infection in males only caused short-term negative effects on sperm morphology,vitality and DNA integrity,but generally recovered to pre-infection levels＞3 months after infection.Due to the limitations of study subjects and sample size,the impact of SARS-CoV-2 infection on male semen quality needs to be further confirmed by long-term large-scale prospective studies.

Objective To optimize the method of combining azomethane oxide(AOM)and dextran sodium sulfate(DSS)to create a colitis-associated colon cancer(CAC)model,and to explore the pathogenesis of the intestinal flora in CAC.Methods Model groups A and B were established by one and two injections of AOM,respectively,combined with free drinking of DSS,and a normal control group was injected intraperitoneally with normal saline combined with purified water(n=10 mice per group).The better modeling scheme was selected by comprehensive evaluation of the disease activity index score,colon length,tumor rate,and mortality.Serum levels of interleukin-6(IL-6),tumor necrosis factor-α(TNF-α),and tumor markers CA199,CEA,and CA724 were detected by enzyme-linked immunosorbent assay.Colon lesions were evaluated by hematoxylin and eosin(HE)staining.Changes in the intestinal microbiota in CAC mice were detected by 16S rDNA high-throughput gene sequencing analysis of mouse feces.Results Both single and enhanced AOM injections combined with DSS induced CAC mice;however,colon growths were larger,more closely arranged,and their morphological size was more consistent in group B compared with group A,with a tumor-formation rate of 100％.IL-6 levels were increased in the model group compared with the normal group(P＜0.05).TNF-α levels were increased in the model group compared with the normal group(P＞0.05).The CA199 and CEA levels were also significantly increased(P＜0.05),but CA724 levels were not.Infiltration of inflammatory cells in the colon detected by HE pathology was accompanied by high-grade intraepithelial tumor-like changes on the surface of the lumen.The diversity and abundance of intestinal bacteria were decreased in CAC mice compared with normal mice:phyla Verrucomicrobiota and Actinobacteriota were significantly increased(P＜0.05),Bacteroidota and Campilobacterota were significantly decreased(P＜0.05).Akkermansia,Prevotellaceae,Ruminococcus,and Bifidobacterium were significantly increased(P＜0.05),and Roseburia,Rikenellaceae_RC9_gut_group,Anaeroplasma,and Muribaculaceae were significantly decreased(P＜0.05).Conclusions Two injections of AOM combined with 1.5％(1.5 g/100 mL)DSS induced CAC model mice with a high colon-tumorigenesis rate,uniform tumor morphology,and low mortality,and may thus be the preferred modeling scheme for pharmacodynamic experiments.Disorders or dysfunction of the intestinal flora may lead to increased permeability,loss of intestinal mucosal barrier function,and the release of enterogenic endotoxins,Resultsing in a sustained inflammatory response,as an indirect or direct cause of CAC pathogenesis.

Objective:To explore the current situation and influencing factors of quality of life of septic patients in intensive care unit (ICU) after discharge, and to provide theoretical basis for clinical early psychological intervention and continuity of care.Methods:A prospective observational study was conducted. The septic patients who were hospitalized in the department of critical care medicine of the Affiliated Hospital of Jining Medical University and discharged with improvement from January 1 to December 31, 2022 were selected as the research objects. The demographic information, basic diseases, infection site, vital signs at ICU admission, severity scores of the condition within 24 hours after ICU admission, various biochemical indexes, treatment process, and prognostic indexes of all the patients were recorded. All patients were assessed by questionnaire at 3 months of discharge using the 36-item short-form health survey scale (SF-36 scale), the activities of daily living scale (ADL scale), and the Montreal cognitive assessment scale (MoCA scale). Multiple linear regression was used to analyze the factors influencing the quality of life of septic patients after discharge from the hospital.Results:A total of 200 septic patients were discharged with improvement and followed up at 3 months of discharge, of which 150 completed the questionnaire. Of the 150 patients, 57 had sepsis and 93 had septic shock. The total SF-36 scale score of septic patients at 3 months of discharge was 81.4±23.0, and the scores of dimensions were, in descending order, role-emotional (83.4±23.0), mental health (82.9±23.6), bodily pain (82.8±23.3), vitality (81.6±23.2), physical function (81.4±23.5), general health (81.1±23.3), role-physical (79.5±27.0), and social function (78.8±25.2). There was no statistically significant difference in the total SF-36 scale score between the patients with sepsis and septic shock (82.6±22.0 vs. 80.7±23.6, P > 0.05). Incorporating the statistically significant indicators from linear univariate analysis into multiple linear regression analysis, and the results showed that the factors influencing the quality of life of septic patients at 3 months after discharge included ADL scale score at 3 months after discharge [ β= 0.741, 95% confidence interval (95% CI) was 0.606 to 0.791, P < 0.001], length of ICU stay ( β= -0.209, 95% CI was -0.733 to -0.208, P = 0.001), duration of mechanical ventilation ( β= 0.147, 95% CI was 0.122 to 0.978, P = 0.012), total dosage of norepinephrine ( β= -0.111, 95% CI was -0.044 to -0.002, P = 0.028), mean arterial pressure (MAP) at ICU admission ( β= -0.102, 95% CI was -0.203 to -0.007, P = 0.036) and body weight ( β= 0.097, 95% CI was 0.005 to 0.345, P = 0.044). Conclusions:The quality of life of patients with sepsis at 3 months after discharge is at a moderately high level. The influencing factors of the quality of life of patients with sepsis at 3 months after discharge include the ADL scale score at 3 months after discharge, the length of ICU stay, the duration of mechanical ventilation, the total dosage of norepinephrine, MAP at ICU admission and body weight, and healthcare professionals should enhance the treatment and care of the patients during their hospitalization based on the above influencing factors, and pay attention to early psychological intervention and continued care for such patients.

ObjectiveTo construct a mouse model of inflammation-associated colorectal cancer （CAC） by using azoxymethane （AOM）/dextran sulfate sodium （DSS） and investigate the effect of Huangqintang on the gut microbiota structure of mice during the occurrence and development of CAC by 16S rRNA gene high-throughput sequencing. MethodA total of 225 C57BL/6J mice were randomized into 5 groups （n=45）： Normal， model， positive drug （mesalazine）， and high （18 g·kg^-1） and low （9 g·kg^-1）-dose Huangqintang. Except those in the normal group， each mouse was injected with 10 mg·kg^-1 AOM on day 1 and day 5 within 1 week and then given 1.5% DSS solution for 7 days， which was then changed to sterile water for 14 days. This process referred to as one cycle， and mice were treated for a total of 3 cycles. On the first day of DSS treatment， mice were administrated with corresponding drugs by gavage， and the normal group and the model group were administrated with pure water by gavage， once a day until the end of the third cycle. The progression of CAC was divided into inflammation， proliferation， and tumorigenesis stages. At the end of each cycle， the body weight and colon length were measured for mice in each group， and the number of colon tumors in mice was recorded. Meanwhile， the disease activity index （DAI） was determined. The serum levels of interleukin-6 （IL-6）， tumor necrosis factor-α （TNF-α）， interleukin-1β （IL-1β）， and carbohydrate antigen-199 （CA199）， a tumor marker in the gastrointestinal tract of mice， were measured by ELISA. Hematoxylin-eosin staining was employed to observe colon lesions. At the same time， 3-5 pellets of fresh feces of mice in the normal group， model group， and high-dose Huangqintang group were collected， from which the fecal DNA of mice was extracted for 16S rRNA gene high-throughput sequencing. ResultCompared with the normal group， the model group showed decreased body weight （P<0.01）， increased DAI， and shortened colon length （P<0.05） at the three stages. Compared with the normal group， the model group showed elevated levels of IL-1β， IL-6， and TNF-α （P<0.05） at the proliferation stage and elevated levels of CA199 at the inflammation， proliferation， and tumorigenesis （P<0.01） stages. Compared with the normal group， the model group presented obvious infiltration of inflammatory cells at the inflammation stage， thickening of the muscle layer and abnormal proliferation of mucosal layer cells at the proliferation and tumorigenesis stages， and final formation of advanced intraepithelial tumor lesions. Compared with the model group， the Huangqintang groups showed no significant improvement in the body weight， decreased DAI score， and increased colon length at the three stages， and the increase of colon length in the tumorigenesis stage was significant （P<0.01）. At the tumorigenesis stage， the administration of Huangqintang inhibited tumor formation and growth， reduced the number of tumors （P<0.01）， lowered the levels of IL-6 （P<0.05， P<0.01）， TNF-α （P<0.05， P<0.01）， and IL-1β at the three stages， and decreased CA199 at the inflammation stage as well as at the proliferation and tumorigenesis stages （P<0.01， P<0.05）. Compared with the model group， the administration of Huangqintang reduced inflammation and abnormal cell proliferation， delaying the occurrence of tumors. Compared with the normal group， the model group showcased decreased alpha and beta diversity and altered structure of gut microbiota at the inflammation， proliferation， and tumorigenesis stages. The administration of Huangqintang adjusted the abundance and diversity of gut microbiota to the normal levels. At the inflammation stage， Huangqintang positively regulated two differential phyla （Firmicutes and Bacteroidetes） and three differential genera （Muribaculaceae， Rikenellaceae_RC9_gut_group， and Flavonifractor） in mice. At the proliferation stage， Huangqintang positively regulated two differential phyla （Bacteroidetes and Patescibacteria） and five differential genera （Muribaculaceae， Rikenellaceae_RC9_gut_group， Candidatus_Saccharimonas， norank_f__UCG-010， and Allobaculum）. At the tumorigenesis stage， Huangqintang positively regulated two differential phyla （Proteobacteria and Patescibacteria） and eight differential genera （Muribaculaceae， Candidatus_Saccharimonas， norank_f_UCG-010， Lachnospiraceae_UCG-006， Allobaculum， Bacteroides， Lachnospiraceae_NK4A136_group， and Flavonifractor） in mice. ConclusionHuangqintang can intervene in the AOM/DSS-induced transformation of inflammation to CAC in mice by correcting inflammation and short-chain fatty acid-related microbiota disorders.

Objective:To investigate the safety and efficacy of posterior apical total intervertebral release (IVR) combined with posterior column osteotomy (PCO) in the treatment of rigid scoliosis.Methods:This study retrospectively analyzed the clinical and radiographic data of 27 patients with rigid scoliosis who underwent posterior total IVR combined with PCO in the apical region from July 2017 to September 2023. There were 10 males and 17 females with an age of 19.3±8.8 years (range 11-48 years). Among them, there were 16 cases of idiopathic scoliosis, 7 cases of neuromuscular scoliosis, 1 case of congenital scoliosis, 1 case of Marfan syndrome with scoliosis, 1 case of neurofibromatosis with scoliosis, and 1 case of osteogenesis imperfecta with scoliosis. The mean Cobb angle of the main curve was 75.4°±13.7° (range 58.7°-110.2°) preoperatively. The mean flexibility of the main curvature is 15.7%±4.7% (range 2.5%-24.3%). Preoperative computer tomography showed that the area of the IVR channel in the convex and concave side of the apical region was 128.1±23.3 mm 2 and 89.5±18.6 mm 2, respectively. The area of the convex IVR was significantly higher than that of the concave IVR. Results:All 27 patients underwent surgery successfully. Total IVR was performed at an average of 3.4±0.7 levels in the apical region. SPO and Ponte osteotomy were performed at 2.7±0.7 and 4.9±1.1 levels, respectively. The mean fusion segment is 11.2±2.0. The operation time, estimated blood loss, and follow-up time were 7.5±0.9 hours (range 6.0-9.8 hours), 1 103.7±845.1 ml (range 300-4 500 ml), and 20.0±14.2 months (range 5-56 months), respectively. The preoperative, postoperative, and final follow-up's mean coronal Cobb angles of the main curve were 75.4°±13.7°, 18.2°±6.5° and 18.6°±6.5°, respectively. The mean correction rate was 75.7%±5.3%. In cases of thoracolumbar kyphosis, the preoperative, postoperative, and final follow-up mean sagittal Cobb angles were 47.2°±4.7°, 22.8°±9.1° and 23.8°±8.9°, respectively. The mean correction rate was 49.5%±18.9%. The mean axial vertebral rotation (AVR) in the IVR region was 24.6°±7.6° preoperatively and was corrected to 11.6°±5.6° postoperatively. The mean correction rate for AVR was 54.0%±11.3%. The coronal, sagittal Cobb angles and AVR postoperatively were significantly lower than those preoperatively ( P<0.001). This case series reported 4 cases of postoperative pleural effusion and 1 case of pulmonary infection, and all of them were cured through conservative treatment. One patient developed incision infection 2 months postoperatively and recovered through debridement surgery. Two patients had proximal junctional kyphosis, one of them underwent revision surgery, and another case was treated with braces. Conclusion:Posterior multi-segment total IVR combined with PCO is a safe and effective surgical procedure for the treatment of rigid scoliosis. The procedure of total IVR was recommended as a supplement for better release of the rigid spine when traditional release methods are not effective.

End-stage liver diseases,such as cirrhosis and liver cancer caused by hepatitis B,are often combined with hepatic encephalopathy(HE);ammonia poisoning is posited as one of its main pathogenesis mechanisms.Ammonia is closely related to autophagy,but the molecular mechanism of ammonia's regulatory effect on autophagy in HE remains unclear.Sialylation is an essential form of glycosylation.In the nervous system,abnormal sialylation affects various physiological processes,such as neural development and synapse formation.ST3 β-galactoside α2,3-sialyltransferase 6(ST3GAL6)is one of the significant glycosyltransferases responsible for adding α2,3-linked sialic acid to substrates and generating glycan structures.We found that the expression of ST3GAL6 was upregulated in the brains of mice with HE and in astrocytes after ammonia induction,and the expression levels of α2,3-sialylated glycans and autophagy-related proteins microtubule-associated protein light chain 3(LC3)and Beclin-1 were upregulated in ammonia-induced astrocytes.These findings suggest that ST3GAL6 is related to autophagy in HE.Therefore,we aimed to determine the regulatory relationship between ST3GAL6 and autophagy.We found that silencing ST3GAL6 and blocking or degrading α2,3-sialylated glycans by way of Maackia amurensis lectin-Ⅱ(MAL-Ⅱ)and neuraminidase can inhibit autophagy.In addition,silencing the expression of ST3GAL6 can downregulate the expression of heat shock protein β8(HSPB8)and Bcl2-associated athanogene 3(BAG3).Notably,the overexpression of HSPB8 partially restored the reduced autophagy levels caused by silencing ST3GAL6 expression.Our results indicate that ST3GAL6 regulates autophagy through the HSPB8-BAG3 complex.

ObjectiveTo investigate the efficacy of Huangqintang on mouse models of colitis-associated colon cancer （CAC） and explore the mechanism of Huangqintang in regulating immune function and inflammatory response， inhibiting abnormal cell proliferation， and delaying or inhibiting CAC formation in CAC. MethodC57BL/6J mice were randomly divided into a normal group， model group， mesalazine group， and high- and low-dose Huangqintang groups according to body weight， with 12 mice in each group. Except for the normal group， the rest of the mice were given two intraperitoneal injections of 10 mg·kg^-1 azomethane （AOM） and allowed to drink 1.5% dextran sodium sulfate （DSS） freely for seven days and water normally for two weeks. Then， two cycles of ''DSS-drinking water'' were repeated. During the administration of DSS， mice in the normal group and model group were given gavage in equal doses of pure water. Mice in the mesalazine group were given 150 mg·kg^-1·d^-1 mesalamine suspension for gavage， and mice in the high- and low-dose Huangqintang groups were given 18 and 9 g·kg^-1·d^-1 Huangqintang for gavage， respectively. Each group was given one dose daily until the end of three cycles. After the intervention， the body weight， colon length， and number of colon tumors in each group were measured， and disease activity index （DAI） scores were performed. The serum contents of interleukin-6 （IL-6）， tumor necrosis factor-α （TNF-α）， interleukin-1β （IL-1β）， interleukin-4 （IL-4）， interleukin-10 （IL-10）， and gastrointestinal tumor marker carbohydrate antigen-199 （CA199） were detected by enzyme linked immunosorbent assay （ELISA）. The colonic lesions were observed by hematoxylin-eosin （HE） staining. The expression of proliferative cell-associated antigen （Ki67） was observed by immunohistochemistry. The expression of T lymphocyte subsets （CD3⁺， CD4⁺， CD8⁺， and CD49b⁺） in mouse plasma was detected by flow cytometry. Fluorescein isothiocyanate-D （FITC-D） content in mouse serum was detected by fluorescent labeling method. The Western blot method was used to detect the expression of Cyclin D₁， cyclin-dependent kinase 2 （CDK2）， cyclin-dependent kinase 4 （CDK4）， and tightly junction-related Occludin and Claudin-1. ResultCompared with the normal group， the body weight of mice in the model group decreased. DAI score increased significantly， and the colon became shorter. Pro-inflammatory factors such as IL-6， TNF-α， and IL-1β increased， and IL-6 and TNF-α were significantly increased （P<0.05）. The inflammatory factor IL-4 （P<0.05） and IL-10 were significantly reduced， and the tumor marker CA199 was significantly increased （P<0.01）. HE staining showed that colon lesions， intestinal mucosal epithelial defects with a large number of inflammatory infiltrates， serious crypt structure damage， and glandular arrangement disorder were observed in the model group. Ki67 positive granules were expressed in large areas of colonic tissue. The serum CD4⁺ and CD4⁺/CD8⁺ of mice in the model group decreased significantly （P<0.05）， and CD8⁺ increased significantly （P<0.05）. The plasma content of FITC-D in the model group was significantly increased （P<0.05）， and the expression of Cyclin D₁， CDK2， and CDK4 proteins in colon tissue was significantly increased （P<0.05， P<0.01）. In addition， the expression of Occludin and Claudin-1 was significantly decreased. Compared with the model group， the body weight of mice in the mesalazine group and the high- and low-dose Huangqintang groups increased. DAI score decreased， and the colon became longer. IL-6， TNF-α， and IL-1β expression decreased （P<0.05， P<0.01）， but there was no significant change in IL-4 and IL-10. The content of CA199 was significantly reduced （P<0.05）， and the colomatoid lesions and inflammatory infiltrates were reduced in the mesalazine group and the Huangqintang group. The crypt structure damage was lighter， and the positive expression of Ki67 was reduced. CD4⁺， CD4⁺/CD8⁺， and CD49b⁺ increased， and the difference was not statistically significant. FITC-D content decreased （P<0.05）. The expression of Cyclin D₁， CDK2， and CDK4 decreased （P<0.05， P<0.01）， and Claudin-1 and Occludin protein expression increased in the high-dose Huangqintang group （P<0.05）. ConclusionHuangqintang has a certain delay and inhibitory effect on AOM/DSS-induced inflammatory cancer transformation， and its mechanism of action may be related to regulating immune function and inflammatory response， inhibiting the release of pro-inflammatory factors， repairing damaged intestinal barriers， inhibiting abnormal proliferation of colon cells， and intervening in the formation and development of CAC colon tumors.

Objective: To study the current situation of sleep quality of senior high school students and analyze the influencing factors, so as to provide a reference for targeted sleep quality improvement of high school students. Methods: A total of 4 793 senior high school students of different grades were recruited from four different regions (Shanghai, Qinghai, Macao, Henan) in 2020. The information of demographic, sleep status, mental factors, living habits and other information were collected by questionnaire. Descriptive statistics and Logistic regression were used to analyze sleep quality and its influencing factors of the senior high school students. Results: About 24.2% of senior high school students reported sleep problems and 71.4% slept less than 7 hours at night. Interpersonal sensitivity, depression, anxiety symptoms, learning, family and interpersonal stress had significant effects on sleep problems in senior high school students( χ 2=1 147.98, 1 228.35, 1 032.54, 371.05, 497.61, 475.39, P <0.01). Drinking coffee and tea, drinking alcohol, smoking (including secondhand smoke), long term use of electronic products, late sleep, bedtime diet, poor sleep environment, parents with sleep problems were the risk factors for sleep disorders of senior high school students, regular exercise was the protective factors for sleep disorders of senior high school students, the influence was significant ( χ 2=160.28, 127.73 , 108.02, 113.27, 470.91, 340.95, 72.02, 155.53, 116.30, 76.96, 28.75, 12.89, P <0.05). Conclusion The sleep quality of senior high school students needs to be improved. Schools and parents should pay attention to the mental health and living habits of senior high school students to help them improve their sleep quality and grow up smoothly.

Objective:To explore the genetic basis for a child featuring tetrahydrobiopterin deficiency and global developmental delay.Methods:Clinical and laboratory examinations were carried out for the child. Genomic DNA of the patient was subjected to high-throughput sequencing to identify genetic variants associated with hyperphenylalaninemia. Candidate variants were verified by Sanger sequencing.Results:The result of blood tandem mass spectrometry showed that the Phenylalanine in the blood was 642.7 μmol/l, and the ratio of Phenylalanine/Tyrosine was 5.42. Analysis of urinary pterin: neopterin 0.09 mmol/mol Cr, biopterin 0.04 mmol/mol Cr, biopterin% 77%, which suggested tetrahydrobiopterin deficiency. The parents of the proband were first cousins. DNA sequencing revealed that the proband has harbored homozygous c. 353A>T variants in exon 2 of the GCH1 gene, for which his great grandmother, grandfather, mother, uncle, father and elder brother were heterozygous carriers with normal phenotype and no clinical symptoms associated with dopa responsive dystonia. Conclusion:The homozygous c. 353A>T variant of the GCH1 gene probably underlay the tetrahydrobiopterin deficiency in this pedigree of consanguineous marriage.