ObjectiveTo investigate the mechanism of Naoxintong capsules on ischemia-reperfusion （I/R） injury in rats based on the changes of pericytes mediated by Ras homolog family member A （RHOA）/Rho-associated coiled-coil containing protein kinase 1 （ROCK1） pathway. MethodsNinety rats （15 rats for each group） were randomly divided into a sham operation group， a model group， a positive control group receiving Ginkgo biloba extract （21.6 mg·kg^-1）， and groups receiving Naoxintong capsules at low， medium， and high doses of 55， 110， and 220 mg·kg^-1 （NXT-L， NXT-M， and NXT-H groups）， respectively. Except for those in the sham operation group， all rats were subjected to transient middle cerebral artery occlusion （tMCAO） to establish the experiment model. Nerve function was assessed using a neurological function score. Cerebral blood flow was detected using a laser speckle contrast imager， and the cerebral infarction rate was calculated using 2，3，5-Triphenyl tetrazolium chloride （TTC） staining. Pathological changes were observed by hematoxylin-eosin （HE） staining and Nissl staining， while pericyte morphology was observed via transmission electron microscopy. Blood-brain barrier destruction was observed by Evans blue staining. Albumin and ischemia-modified albumin levels were measured using assay kits. Real-time fluorescence quantitative polymerase chain reaction （Real-time PCR） and Western blot were used to detect the mRNA and protein expression levels of RHOA， ROCK1， platelet-derived growth factor receptor β （PDGFRB）， α-smooth muscle actin （α-SMA）， tight junction protein （ZO-1）， matrix metalloproteinase-2 （MMP-2）， and matrix metalloproteinase-9 （MMP-9）. ResultsCompared with the sham operation group， the model group exhibited decreased neurological function scores， higher percentage reduction in blood flow， and increased cerebral infarction rates （P<0.01）. Additionally， cortical neuronal nucleus shrinkage， edema， a decreased number of Nissl bodies， reduced pericyte area， elevated albumin content in the cortex （P<0.05）， and increased ischemic modified albumin levels （P<0.01） were observed. The mRNA and protein expression levels of RHOA， ROCK1， PDGFRB， α-SMA， MMP-2， and MMP-9 were increased （P<0.01）， while those of ZO-1 were decreased. Compared with the model group， all treatment groups showed improved neurological function scores， lower percentage reduction in blood flow， reduced cerebral infarction rates （P<0.01）， alleviated cortical histological changes， increased number of Nissl bodies， expanded pericyte area， decreased albumin content in the cortex， and reduced ischemia-modified albumin levels （P<0.01）. The mRNA and protein expression levels of RHOA， ROCK1， PDGFRB， α-SMA， MMP-2， and MMP-9 were decreased （P<0.01）， while those of ZO-1 were increased. Among the treatment groups， the NXT-M group showed the most pronounced improvement in cerebral I/R injury. ConclusionNaoxintong capsules can restore cerebral blood supply， reduce microcirculation disturbance， and protect blood-brain barrier in rats with I/R injury. Its mechanism of action may be related to the inhibition of the RHOA/ROCK1 signaling pathway and reduced pericyte contraction.

ObjectiveTo investigate the mechanism of Naoxintong capsules on ischemia-reperfusion （I/R） injury in rats based on the changes of pericytes mediated by Ras homolog family member A （RHOA）/Rho-associated coiled-coil containing protein kinase 1 （ROCK1） pathway. MethodsNinety rats （15 rats for each group） were randomly divided into a sham operation group， a model group， a positive control group receiving Ginkgo biloba extract （21.6 mg·kg^-1）， and groups receiving Naoxintong capsules at low， medium， and high doses of 55， 110， and 220 mg·kg^-1 （NXT-L， NXT-M， and NXT-H groups）， respectively. Except for those in the sham operation group， all rats were subjected to transient middle cerebral artery occlusion （tMCAO） to establish the experiment model. Nerve function was assessed using a neurological function score. Cerebral blood flow was detected using a laser speckle contrast imager， and the cerebral infarction rate was calculated using 2，3，5-Triphenyl tetrazolium chloride （TTC） staining. Pathological changes were observed by hematoxylin-eosin （HE） staining and Nissl staining， while pericyte morphology was observed via transmission electron microscopy. Blood-brain barrier destruction was observed by Evans blue staining. Albumin and ischemia-modified albumin levels were measured using assay kits. Real-time fluorescence quantitative polymerase chain reaction （Real-time PCR） and Western blot were used to detect the mRNA and protein expression levels of RHOA， ROCK1， platelet-derived growth factor receptor β （PDGFRB）， α-smooth muscle actin （α-SMA）， tight junction protein （ZO-1）， matrix metalloproteinase-2 （MMP-2）， and matrix metalloproteinase-9 （MMP-9）. ResultsCompared with the sham operation group， the model group exhibited decreased neurological function scores， higher percentage reduction in blood flow， and increased cerebral infarction rates （P<0.01）. Additionally， cortical neuronal nucleus shrinkage， edema， a decreased number of Nissl bodies， reduced pericyte area， elevated albumin content in the cortex （P<0.05）， and increased ischemic modified albumin levels （P<0.01） were observed. The mRNA and protein expression levels of RHOA， ROCK1， PDGFRB， α-SMA， MMP-2， and MMP-9 were increased （P<0.01）， while those of ZO-1 were decreased. Compared with the model group， all treatment groups showed improved neurological function scores， lower percentage reduction in blood flow， reduced cerebral infarction rates （P<0.01）， alleviated cortical histological changes， increased number of Nissl bodies， expanded pericyte area， decreased albumin content in the cortex， and reduced ischemia-modified albumin levels （P<0.01）. The mRNA and protein expression levels of RHOA， ROCK1， PDGFRB， α-SMA， MMP-2， and MMP-9 were decreased （P<0.01）， while those of ZO-1 were increased. Among the treatment groups， the NXT-M group showed the most pronounced improvement in cerebral I/R injury. ConclusionNaoxintong capsules can restore cerebral blood supply， reduce microcirculation disturbance， and protect blood-brain barrier in rats with I/R injury. Its mechanism of action may be related to the inhibition of the RHOA/ROCK1 signaling pathway and reduced pericyte contraction.

Objective To explore the detection rate and distribution characteristics of gene mutations in pa-tients with non-small cell lung cancer,and to analyze their relationship with clinicopathological characteristics.Methods The genetic testing results and clinicopathological data of 213 patients with non-small cell lung cancer who underwent high-throughput genetic testing at the Gene Detection Center,Changchun Cancer Hos-pital from April 2020 to December 2023 were collected.High-throughput sequencing technology was used to detect mutations in 26 genes,and the relationship between the mutation frequency and its distribution and the clinicopathological characteristics of patients was analyzed.Results Among 213 patients with non-small cell lung cancer,192 cases(90.14％)had at least one gene mutation detected.Among them,the genes with rela-tively high mutation frequencies were TP53(60.56％),EGFR(46.48％),KRAS(14.55％),ALK fusion(11.74％),and PIK3CA(8.92％).There were 132 cases(30.28％)of evidence level for Class 1 drugs and 11 cases(2.52％)of evidence level for Class 2 drugs.The incidence of EGFR gene mutations was higher in women,non-smokers and patients with lung adenocarcinoma(P＜0.05).TP53 mutations usually occured in women,smokers and patients with stage Ⅳ.ALK mutations were more common in young patients,while KRAS mutations were more frequently seen in male smokers.Conclusion Analyzing the distribution charac-teristics of gene mutations in non-small cell lung cancer and their relationship with clinicopathological charac-teristics can provide a scientific basis for further optimizing genetic testing for patients with non-small cell lung cancer and offer guidance for clinical treatment.

OBJECTIVE To optimize the preparation technology of the volatile oil microcapsules of Cinnamomi Ramulus and Angelicae Sinensis Radix, characterize the microcapsules prepared after the formulation optimization and study pharmacokinetics characteristics in rats. METHODS Spray drying was used to prepare the volatile oil microcapsules of Cinnamomi Ramulus and Angelicae Sinensis Radix. The preparation process was optimized by Box-Behnken response surface method with the drug loading and encapsulation efficiency as indexes. The microcapsules were characterized by Fourier transform infrared spectroscopy(FT-IR) and scanning electron microscopy(SEM). Blood samples were collected after gastric administration at a dose of 100 mg·kg-1. HPLC method was adopted in the plasma concentration determination, and pharmacokinetics behavior in vivo was also compared. RESULTS The optimal formulation: core material-capsules material ratio was 1∶1.7, capsules material concentration was 10.25%, sodium starch octenyl succinate∶maltodextrin was 3.8∶1, high pressure homogenization pressure was 20 MPa and the inlet temperature of spray drying was 185 ℃. Under the optimized conditions, the drug loading was (18.94±1.09)% and the encapsulation rate was (96.03±2.91)%, respectively. The results of FT-IR and SEM showed that the essential oils had been successfully coated in the capsule wall material. The microcapsules were basically spherical in shape, with concave surface but no obvious cracks. The main pharmacokinetic parameters such as tmax, t1/2, CL, AUC0-t, k10,k12, and k21 of microcapsules had significant difference of the volatile oil(P<0.05 or P<0.01). Microcapsules effectively prolonged the circulation time of volatile oil in the body, promoted the absorption of drugs in the body and the oral bioavailability was enhanced to 2.62 times. CONCLUSION The model established by the Box-Behnken design-response surface method can be used to optimize the formulation of volatile oil microcapsules of Cinnamomi Ramulus and Angelicae Sinensis Radix with great prediction effect. The microcapsules prepared by the optimized process has good drug loading properties and the bioavailability of volatile oil is increased.

Objective:To analyze the phenotypes and genetic etiology of microcephaly- seizures-development delay (MCSZ) syndrome.Methods:The patient was diagnosed with MCSZ syndrome in June 2018 at Shenzhen Maternity and Child Healthcare Hospital. She was the couple's first child, and the mother conceived a second child in 2020. The clinical data of the proband were retrospectively analyzed, and the bioinformatics analysis and whole-exome sequencing (WES) were performed on the proband and her parents to identify the pathogenic variants, which were further validated using Sanger sequencing. The prenatal genetic diagnosis of the second fetus was performed following the molecular diagnosis of the proband was confirmed. The clinical manifestations and pathogenesis of MCSZ syndrome were summarized by reviewing related literature.Results:(1) Case report: The patient, an eight-month-old girl, was admitted to our hospital due to microcephaly and repeated seizures. Another clinical characteristic was mental retardation. Auditory evoked potential detected moderate impairment of the left auditory nerve pathway. WES showed a compound heterozygous variation in the PNKP gene of the proband. Moreover, the pathogenic variation, c.199-10_203delinsTCTGAGGGGT, was inherited from the father, and the likely pathogenic variation, c.1505C>T(p.P502>L), was inherited from the mother, which was both de novo mutations. The compound heterozygous variation in the PNKP gene was considered genetic etiology based on the genetic testing and clinical features. Prenatal diagnosis showed that the second fetus did not inherit the PNKP gene variants from the parents and the couples chose to continue the pregnancy. A girl was born, and her psychomotor development and occipitofrontal size circumference were normal at 13 months old. (2) Literature review: 39 MCSZ syndrome cases were retrieved, including the present case and 38 cases from 12 relevant literature. The clinical characteristics were microcephaly (91.7%, 33/36), seizures (88.2%, 30/34), development delay (96.4%, 27/28), hyperactivity (25.6%, 9/39), gastroesophageal reflux (10.3%, 4/39), and hearing loss (7.7%, 3/39). Most patients' first onset of epilepsy was in infancy (96.3%, 26/27). Cranial MRI examination showed brain dysplasia in 31 cases (91.2%, 31/34). Conclusions:When the fetal head circumference is smaller than normal and is progressively reduced combined with postnatal microcephaly, epilepsy, developmental retardation, hyperactivity disorder, gastroesophageal reflux, and hearing loss, MCSZ syndrome should be considered. The prognosis varies widely, and genetic testing facilitates the early diagnosis and genetic counseling of MCSZ syndrome.

Two pedigrees are reported here including two siblings and a boy who were diagnosed with Aicardi-Goutières syndrome type 3 (AGS3) caused by compound heterozygous variation of RNASEH2C gene. Prenatal gene diagnosis was performed when their mothers were pregnant again. All three cases presented with epilepsy, microcephaly, muscular hypertonia and severe language, motor and mental retardation. In pedigree 1, genetic analysis showed compound heterozygous variants of c.194G>A (p.Gly65Asp) and c.434G>T (p.Arg145Leu) in the RNASEH2C gene of proband 1 and her younger brother, which were inherited from their mother and father respectively. While in pedigree 2, c.194G>A(p.Gly65Asp) and c.227C>T(p.Pro76Leu) compound heterozygous variants in the RNASEH2C gene were found in proband 2, which were inherited from his father and mother, respectively. Diagnosis of AGS3 was confirmed in these three cases based on their medical history and the testing results. The mothers from the two families underwent prenatal diagnosis in their subsequent pregnancy, and the variation only inherited from the mothers was detected, suggesting that the two fetuses are carriers. Both families chose to continue the pregnancy and delivered at full-term. No growth or development abnormalities were reported in the children during a one-year follow-up.

Objective:To summarize the genetic diagnosis of two fetuses with clinically suspected Bardet-Biedl syndrome (BBS) and to provide information for genetic counseling and prenatal diagnosis of BBS.Methods:Case one had prenatal care on October 2018 in Shenzhen Maternity and Child Healthcare Hospital and was clinically suspected of fetal BBS as bilateral renal parenchyma echo enhancement as well as polydactyly (six toes on each foot) were shown on ultrasonic examination at 18 +1 gestational weeks. Case two was another suspected fetal BBS for enlarged kidneys with echo enhancement as well as polydactyly (six fingers and toes on each hand and foot) on ultrasonic examination at 26 +4 gestational weeks on August 2016 and the parent requested for termination. Parents of both cases requested for genetic analysis. Amniotic fluid sample was obtained in case one at 19 +6 weeks through amniocentesis, and umbilical cord specimen of case two and peripheral blood samples of the parents were collected. Genetic analysis of the fetuses and their parents was performed using exon capture and next-generation sequencing and the results were validated using Sanger sequencing. Results:Case one carried paternally inherited c.718G>A (p.Gly240Ser) (possible pathogenic) mutation and maternally inherited c.497C>A(p.Ala166Asp) (possible pathogenic) mutation in BBS7 gene. While one paternally inherited mutation c.1002delT(p.N335Ifs*47) (pathogenic) and one maternally inherited heterozygous mutation c.728G>A (p.Cys243Tyr) (possible pathogenic) were identified in BBS7 gene of case two. The three unreported missense mutations were predicted to be harmful by bioinformatics software and the mutation sites were conservative after comparing with multiple species-based protein sequences. Conclusions:Enlarged kidneys with echo enhancement and polydactyly may indicated a BBS fetus caused by BBS7 gene mutation. Whole exome sequencing could provide relevant information for prenatal diagnosis and genetic counseling in these cases.

Objective To investigate the method for preparing oridonin-single-walled carbon nanotubes ( ORI-SWCNTs) nanocomposite and study its adsorption kinetics. Methods ORI-SWCNTs nanocomposite was prepared by using the method of solution mixing. The synthesized ORI-SWCNTs nanocomposite was characterized by using Laser particle size analyzer, Fourier transform infrared, DSC analysis, powder X-ray diffraction and electron microscopy techniques. Results The encapsulation efficiency and loading capacity of ORI in SWCNTs-COOH nanocarrier was estimated to be about (70.23±2.1) %and (27.29±1.2) %, respectively. The Zeta potential was (-34.29±1.2) mV, partical size was about (458±18) nm. The absorption of ORI on SWCNTs-COOH could be explained by pseudo-second-order model. Conclusion The established preparation process of ORI-SWCNTs nanocomposite by solution mixing is feasible, with higher loading efficiency and encapsulate efficiency..

Objective To study the effect of fascia parotideomasseterica reconstruction in the prevention of Frey syndrome.Methods 45 patients who suffered parotidectomy with parotid fascia maintained and reconstructed were followed-up,the OccurrenCe rate of Frey syndrome wasobserved.Results The occurrence rate of the subjective symptom8 with Frey syndrome Was 6.7%(3/45)6 montes after surgery;the occurrence rate of Frey syndrome was 11.1%(5/45)by Minor test review 1 year after surgery;the remaining 41 cases were non-induced reactlons.Condusion Fascia parotidcomassetefica could reduce the development of Frey syndrome in the parotid gland surgery in pafients with benign tumors.