Effect of Qiwei Baizhusan on Cognitive Dysfunction in Rats with Diabetic Encephalopathy Based on PI3K/Akt/GSK-3β Signaling Pathway / 中国实验方剂学杂志

ObjectiveTo observe the therapeutic effect of Qiwei Baizhusan（QWBZS） on diabetic encephalopathy（DE） rat model， and to explore the possible mechanism of QWBZS in the treatment of DE based on phosphatidylinositol 3-kinase（PI3K）/protein kinase B（Akt）/glycogen synthase kinase-3β（GSK-3β） signaling pathway. MethodForty-eight SPF male Wistar rats were randomly divided into blank group（8 rats） and high-fat diet group（40 rats）. After 12 weeks of feeding， rats in the high-fat diet group were intraperitoneally injected with 35 mg·kg-1 of 1% streptozotocin（STZ） for 2 consecutive days to construct a DE model， and rats in the blank group were injected with the same amount of sodium citrate buffer. After successful modeling， according to blood glucose and body weight， model rats were randomly divided into model group， low， medium and high dose groups of QWBZS（3.15， 6.3， 12.6 g·kg-1）， combined western medicine group（metformin+rosiglitazone， 0.21 g·kg-1）， with 6 rats in each group. The administration group was given the corresponding dose of drug by gavage， and the blank group and the model group were given an equal volume of 0.9% sodium chloride solution by gavage， 1 time/day for 6 weeks. Morris water maze was used to detect the spatial memory ability of DE rats. Fasting insulin （FINS） level was detected by enzyme-linked immunosorbent assay（ELISA） and insulin resistance index（HOMA-IR） was calculated. Hematoxylin-eosin（HE） staining was used to observe the morphological changes of hippocampus in rats， ELISA was used to detect the indexes of oxidative stress in hippocampal tissues， real-time fluorescence quantitative polymerase chain reaction（Real-time PCR） was used to detect mRNA expression levels of PI3K， Akt， nuclear transcription factor-κB（NF-κB）， tumor necrosis factor-α（TNF-α） and interleukin-1β（IL-1β） in hippocampus， and Western blot was used to detect the protein expression of PI3K， Akt， phosphorylated（p）-Akt， GSK-3β and p-GSK-3β in hippocampus of rats. ResultCompared with the blank group， FINS and HOMA-IR values of the model group were significantly increased（P<0.01）， the path of finding the original position of the platform was significantly increased， and the escape latency was significantly prolonged（P<0.01）， the morphology of neuronal cells in hippocampal tissues was disrupted， the levels of reactive oxygen species（ROS） and malondialdehyde（MDA） in hippocampus of rats were increased， and the activity of superoxide dismutase（SOD） was decreased（P<0.05， P<0.01）， mRNA expression levels of PI3K and Akt were decreased（P<0.01）， mRNA expression levels of NF-κB， TNF-α and IL-1β were increased（P<0.05， P<0.01）， the protein expression levels of PI3K， p-Akt and p-GSK-3β were significantly decreased， and the protein expression of GSK-3β was significantly increased（P<0.01）. Compared with the model group， the FINS and HOMA-IR values of the medium dose group of QWBZS and the combined western medicine group were significantly decreased（P<0.01）， the path of finding the original position of the platform and the escape latency were significantly shortened（P<0.01）， the hippocampal tissue structure of rats was gradually recovered， and the morphological damage of nerve cells was significantly improved， the contents of ROS and MDA in hippocampus of rats decreased and the level of SOD increased（P<0.01）， the mRNA expression levels of PI3K and Akt were increased（P<0.01）， and the mRNA expression levels of NF-κB， TNF-α and IL-1β were decreased （P<0.05， P<0.01）， the protein expression levels of PI3K， p-Akt and p-GSK-3β were significantly increased（P<0.01）， and the expression of GSK-3β was significantly decreased（P<0.01）. ConclusionQWBZS can alleviate insulin resistance in DE rats， it may repair hippocampal neuronal damage and improve learning and cognitive ability of DE rats by activating PI3K/Akt/GSK-3β signaling pathway.

Value of CT radiomic features in differential diagnosis of lung metastases / 国际肿瘤学杂志

Objective:To distinguish lung metastases of different origin by constructing a classification model according to CT radiomics features.Methods:A total of 226 patients with lung metastases of gastric cancer, breast cancer and kidney cancer attending Chongqing Red Cross Hospital from January 2015 to July 2020, with a total of 402 metastases, were randomly divided into a training cohort (training set, 136 patients, 280 metastases) and a validation cohort (validation set, 90 patients, 122 metastases) by the hold-out method. In addition, 68 patients with lung metastases (138 lung metastases in total) attending Chongqing Red Cross Hospital from August 2020 to April 2022 were matched as an external test cohort (test set). Region of interest segmentation was performed by two experienced radiologists independently and manually without clinical information to construct the model by using LASSO screening for the best radiomic features. Support vector machine (SVM) and random forest (RF) were selected to build dichotomous and trichotomous models respectively. The receiver operating characteristic curve was used to evaluate the classification efficiency of both models.Results:There were no statistically significant differences in age ( t=-0.06, P=0.534), gender ( χ2<0.01, P=0.961) and number of lung metastases ( χ2=0.71, P=0.703) between the validation and test sets. A total of 792 radiomic features were extracted, 703 of which had good agreement (intraclass correlation coefficient≥0.75), while 89 features being excluded for having poor agreement (intraclass correlation coefficient<0.75). The dichotomous model (SVM) screened 28 (lung metastases from gastric cancer vs. lung metastases from breast cancer), 25 (lung metastases from gastric cancer vs. lung metastases from kidney cancer) and 34 (lung metastases from kidney cancer vs. lung metastases from breast cancer) features, respectively; the trichotomous model (RF) screened 20 features (three types of lung metastases), in which Short Run Emphasis and Inverse Variance were significantly higher in lung metastases from kidney cancer than in the other two types, correlation was higher in lung metastases from gastric cancer than in the other two types, and there was no significant difference in the sphericity of the three lung metastases. For the dichotomous model, in the validation set, the area under the curve (AUC) of the 28 features selected to distinguish gastric cancer lung metastases from breast cancer lung metastases was 0.81, the AUC of the 25 features distinguishing gastric cancer lung metastases from kidney cancer lung metastases was 0.86, and the AUC of the 34 features distinguishing kidney cancer lung metastases from breast cancer lung metastases was 0.92, and the AUCs of the test set were 0.80, 0.79 and 0.86 respectively. For the trichotomous model, the AUC for predicting lung metastases from gastric cancer, breast cancer and kidney cancer in the validation set were 0.85, 0.82 and 0.91 respectively, and both macroscopic and microscopic AUC were 0.85; In the test set, the AUC for predicting lung metastases from gastric cancer, breast cancer, and kidney cancer were 0.77, 0.86 and 0.84 respectively, and both macroscopic and microscopic AUC were 0.81. Conclusion:The SVM and RF models based on CT radiomic features are helpful in distinguishing lung metastases derived from gastric cancer, breast cancer and kidney cancer.

Impact of lead oxide nanoparticle exposure on the polarization of microglia cells in mouse hippocampus / 中国职业医学

Objective To investigate the effect of exposure to lead oxide nanoparticles (PbO NPs) on the polarization of microglia in mouse hippocampus. Methods i) Specific pathogen-free male C57 mice were randomly divided into control group, low-, medium- and high-dose groups, with 10 mice in each group. Mice in these three dose groups were intraperitoneally injected with PbO NPs suspension at doses of 5, 10 and 20 mg/kg per day, respectively, and mice in the control group were intraperitoneally injected with the same volume of 0.9% sodium chloride solution, five days per week for four weeks. ii) BV-2 cells were treated with PbO NPs at doses of 0.0, 2.5, 5.0 and 10.0 mg/L for 24 hours. iii) BV-2 cells were randomly divided into control group, PbO NPs group and triggering receptor expressed on myeloid cells 2 (TREM2) high expression + PbO NPs group. The cells in the control group received no treatment. The cells in PbO NPs group were exposed to 10.0 mg/L PbO NPs suspension for 24 hours. Cells in TREM2 high expression + PbO NPs group were transfected with Trem2 high expression plasmid, and then exposed to 10.0 mg/L PbO NPs suspension for 24 hours. iv) The mRNA expression of M1 markers ［nitric oxide synthase (iNos), cyclooxygenase 2 (Cox2), chemokine receptor 7 (Ccr7)］, M2 markers ［arginin-1 (Arg-1), transforming growth factor-β (Tgf-β), chemokine receptor 2 (Ccr2)］ and Trem2 of microglia was detected by real-time fluorescent quantitative polymerase chain reaction. The protein expression of iNOS, ARG-1 and TREM2 was detected by Western blotting. Results i) During the experiment, there was no significant difference in body weight of mice among these four groups (P>0.05). The relative expression of Cox2 and Ccr7 mRNA in the hippocampus of the mice increased in the low-dose group and the iNos, Cox2 and Ccr7 mRNA increased in the medium- and high-dose groups, compared with the control group (all P<0.05). The relative mRNA expression of Tgf-β in the hippocampus of the mice of low-dose group and Arg-1, Tgf-β and Ccr2 in the medium- and high-dose groups was decreased compared with the control group (all P<0.05). The mRNA relative expression of iNos, Cox2 and Ccr7 was increased (all P<0.05), while the mRNA relative expression of Arg-1, Tgf-β and Ccr2 was decreased (all P<0.05) in the hippocampus of the mice of high-dose group compared with the low-dose group. The relative expression of Trem2 mRNA and TREM2 protein in the hippocampus of mice of the medium- and high-dose groups was lower than those in the control group (all P<0.05). The relative expression of Trem2 mRNA and TREM2 protein in the hippocampus of mice of the high dose group was lower than those in the low- and the medium-dose groups (all P<0.05). With the increase of PbO NPs exposure dose, the relative expression of iNOS protein in hippocampus tissues of mice increased (P<0.01), and the relative expression of ARG-1 protein decreased (P<0.01). ii) With the increase of PbO NPs exposure dose, the relative expression of iNOS protein increased (P<0.01), and the relative expression of ARG-1 protein decreased (P<0.01) in BV-2 cells. The relative expression of iNOS protein in BV-2 cells of PbO NPs group and TREM2 high expression + PbO NPs group was increased (all P<0.05), and the relative expression of ARG-1 protein decreased (all P<0.05) compared with the control group. The relative expression of iNOS protein decreased (P<0.05), and the relative expression of ARG-1 protein increased (P<0.05) in BV-2 cells of TREM2 high expression + PbO NPs group compared with the PbO NPs group. Conclusion Exposure to PbO NPs could increase the M1 polarization and decrease the M2 polarization of microglia, with a dose-effect relationship. The M1 polarization of microglia decreased and M2 polarization increased after overexpression of Trem2 gene. The regulation of microglia polarization by TREM2 may be involved in the neurotoxic effects of PbO NPs.

Detection of ATP2C1 gene mutations in a family with generalized familial benign chronic pemphigus / 中华皮肤科杂志

A 60-year-old female proband presented with recurrent erythema, blisters and erosions all over the body for 30 years, which had been aggravated 10 days prior to the presentation. Skin examination showed erythematous swelling of the bilateral eyelids with scattered dark red crusts, scattered erythema and erosions on the nasolabial folds and chin, large areas of erythema and erosions on the neck, bilateral axillae, left cubital fossa, perineum and perianal area, accompanied by bright red granulation tissues and positive Nikolsky′s sign. The proband had two sons, both of whom occasionally presented with erythema and erosions on the axillae and groin, and had not been diagnosed or treated. Blood samples were collected from the proband and her two sons, and genomic DNA was extracted and subjected to whole-exome sequencing. A heterozygous deletion mutation c.955_957del (p.A319del) was identified in the ATP2C1 gene in the proband and her two sons, which had not been previously reported. The patient was finally diagnosed with generalized familial benign chronic pemphigus.

Gene mutation analysis in a Chinese pedigree with autosomal dominant Waardenburg syndrome / 中华皮肤科杂志

Objective:To report a Chinese pedigree with autosomal dominant Waardenburg syndrome, and to identify causative gene mutations.Methods:Clinical data and peripheral blood samples were collected from the proband and her parents. Genomic DNA was extracted, gene mutations were detected through a next-generation skin-targeted sequencing panel, and Sanger sequencing was performed to verify causative mutations.Results:The proband clinically presented with irregular white patches on the abdomen and lower limbs, moderate to severe sensorineural deafness in the right ear, and iris heterochromia in both eyes. The proband′s mother presented with iris heterochromia in both eyes, epicanthus, early canities and thick eyebrows. In the family, both the proband and her mother were diagnosed with Waardenburg syndrome. A causative frameshift mutation c.976-977delinsT (p.Thr327Profs*54) was identified in both the proband and her mother, which caused the AG to TT base substitution at positions 976 - 977 in the coding region of exon 7 of the PAX3 gene, resulted in a frameshift from the amino acid position 327 to 54 in the PAX3 protein (threonine was substituted by proline at amino acid position 327). The proband′s father showed a normal phenotype, and his genetic test results were negative.Conclusion:The novel frameshift mutation c.976-977delinsT (p.Thr327Profs*54) in the PAX3 gene may contribute to the clinical phenotype of the patients with Waardenburg syndrome in the family.

Variation of COL7A1 gene in dystrophic epidermolysis bullosa pruriginosa / 中华医学遗传学杂志

OBJECTIVE@#To perform gene mutation analysis in a Chinese pedigree with dystrophic epidermolysis bullosa pruriginosa (DEB-Pr), and explore phetotype, genotype, and genotypes-phenotypes relationship of DEB-Pr.@*METHODS@#Potential variants of the COL7A1 gene were detected by skin targeted sequencing panel and verified by Sanger sequencing. The pathogenicity of the variation was analyzed.@*RESULTS@#Compound heterozygous variants, c.4128delT and c.8234G>A, were detected in the COL7A1 gene of the two patients. The c.4128delT(p.Pro1376fs) variant was derived from their mother and unreported previously. According to the American College of Medical Genetics and Genomics Standards and Guidelines, it was suggested to be a pathogenic mutation. The c.8234G>A(p.Arg2745Gln) variant was derived from their father, and possibly is a pathogenic variation.@*CONCLUSION@#In this study, the compound heterozygous variants of c.4128delT(p.Pro1376fs) and c.8234G>A(p.Arg2745Gln) of the COL7A1 gene probably underlies the disease in this patient and his sister. And our study expands the database on mutations of DEB-Pr.

Mutation analysis of the ADAR gene in two cases of dyschromatosis symmetrica hereditaria and a survey of their families / 中华皮肤科杂志

Objective:To investigate two Chinese pedigrees with dyschromatosis symmetrica hereditaria (DSH) , and to analyze gene mutations in the pedigrees.Methods:Clinical data were collected from two probands with DSH and other family members in their pedigrees. Peripheral blood samples were obtained from the two probands, their parents and 100 unrelated healthy controls. Gene mutations were detected by using a skin-targeted sequencing panel, and then verified by Sanger sequencing.Results:Case 1, an 18-year-old male patient, presented with millet-sized hyperpigmented and hypopigmented macules scattered on the dorsum of both hands and feet at the age of 5 years, and his mother had similar manifestations. A novel heterozygous frameshift mutation c.1970dupT (p.F657fs) was identified in exon 5 of the ADAR gene in case 1 and his mother, but not found in his father. Case 2, an 8-year-old male patient, presented with mottled rice- to soybean-sized brown hyperpigmented macules and hypopigmented macules on the face and neck, lower back, buttocks, lower limbs, as well as hands and feet, and his father presented with similar manifestations. A known heterozygous frameshift mutation c.2433_2434delAG (p.T811fs) was identified in exon 7 of the ADAR gene in case 2 and his father, but not found in his mother. Neither of the two mutations was identified in the 100 unrelated healthy controls.Conclusion:In this study, a novel mutation c.1970dupT (p.F657fs) in the ADAR gene was identified in a patient with DSH.

Rare subtypes of epidermolysis bullosa: three case reports and their pedigree analysis / 中华皮肤科杂志

Objective:To report 3 cases of rare subtypes of hereditary epidermolysis bullosa.Methods:Clinical data were collected from the probands and their relatives, whole-exome sequencing was performed to screen disease-causing mutations in the probands, and Sanger sequencing or qPCR was conducted to verify the mutations in patients and their relatives.Results:Case 1 mainly presented with linear red scars on the back, and the proband, her mother with similar clinical manifestations and her asymptomatic daughter all carried a mutation c.4573G>A (p.Gly1525Arg) in the COL7A1 gene. Case 2 presented with generalized reticular pigmentation all over the body and occasional blisters restricted to the hand and foot, and carried a de novo mutation c.74C>T (p.Pro25Leu) in the KRT5 gene. Case 3 presented with pigmentation abnormalities mainly located at the sun-exposed sites and incomplete syndactyly of the left hand, and carried homozygous deletion mutations in exons 2-6 of the FERMT1 gene, which were inherited from her asymptomatic parents. Case 1 was diagnosed with dominant dystrophic epidermolysis bullosa pruriginosa, case 2 was diagnosed with epidermolysis bullosa simplex with mottled pigmentation, and case 3 was diagnosed with Kindler epidermolysis bullosa. Conclusion:The clinical manifestations of epidermolysis bullosa vary greatly, and gene detection is very important for confirmation of diagnosis of its rare types.

Effect of HaCaT cells with CRISPR-Cas9-induced KRT5 mutation on co-cultured human melanocytes / 中华皮肤科杂志

Objective:To investigate the effect of KRT5 knockdown in keratinocytes on melanin content in co-cultured melanocytes, and to explain mechanisms underlying formation of hyperpigmented lesions in reticulate pigmented anomaly of the flexures (Dowling-Degos disease, DDD) .Methods:HaCaT cells with heterozygous mutations in the KRT5 gene were obtained by using clustered regularly interspaced short palindromic repeats (CRISPR) -CRISPR-associated protein 9 (Cas9) technology (experimental group) , and HaCaT cells transfected with non-targeting single guide RNA:Cas9 protein complex served as control group, both of which were in vitro co-cultured with primary human melanocyte cells (HEMn) separately. Immunofluorescence study was conducted to determine the expression of cytokeratin and melanosomes in co-cultured cells; melanin content was detected in melanocytes in different co-culture groups, which were obtained by differential trypsinization. Immunohistochemical study was performed to determine the expression of melanocyte-specific premelanosome protein 17 (Pmel17) in skin lesions in a patient with DDD carrying a KRT5 mutation and normal skin tissues in a healthy control. Results:Sanger sequencing showed a heterozygous mutation (c.1delA) at the initiation codon of exon 1 of the KRT5 gene in HaCaT cells in the experimental group, but no mutation in the KRT5 gene in the control group. Western blot analysis showed that the KRT5 protein expression was significantly lower in the experimental group (0.60 ± 0.05) than in the control group (1.00 ± 0.00, t = 32.38, P = 0.001) . Compared with the co-culture system in the control group, the number of Pmel17-labeled melanosomes markedly increased with the melanin content elevated by 52.5% ( t = -3.48, P = 0.025) in the HEMn cells co-cultured with HaCaT cells in the experimental group. Immunohistochemical study showed that the Pmel17 expression increased in the skin lesions in the DDD patient with KRT5 mutation compared with the normal skin tissues in the healthy control. Conclusion:The effect of HaCaT cells with CRISPR-Cas9-induced KRT5 mutation on the co-cultured HEMn melanocytes was verified by the successfully established in vitro co-culture system, which provides a primary cell model for further studies on interaction mechanisms between keratinocytes and melanocytes, and on pathogenesis of skin pigmentation abnormalities.

Gene mutation analysis in a Chinese pedigree with autosomal recessive woolly hair / 中华皮肤科杂志

Objective:To identify causative genes for autosomal recessive woolly hair (ARWH) in a family.Methods:Clinical data were collected from two patients and other family members in a Chinese pedigree of Han nationality with ARWH. Peripheral blood samples were obtained from the two patients, their unaffected parents and 100 unrelated healthy individuals, and DNA was extracted from the blood samples. A next-generation skin-targeted sequencing panel was used to detect gene mutations in the patients, and Sanger sequencing was performed to verify the sequencing results. The function of protein encoded by the mutant gene was predicted.Results:Two missense mutations c.530T>G (p.Leu177Arg) and c.736T>A (p.Cys246Ser) were both identified in the LIPH gene of the two patients, which were inherited from their father and mother respectively. Neither of the two mutations was identified in the 100 unrelated healthy controls. Interspecies sequence alignment showed that leucine at amino acid position 177 and cysteine at amino acid position 246 of the protein encoded by the LIPH gene were highly evolutionarily conserved. As SIFT and Polyphen-2 softwares showed, the mutations c.530T>G (p.Leu177Arg) and c.736T>A (p.Cys246Ser) were both predicted to be detrimental variations.Conclusion:Two missense mutations c.530T>G (p.Leu177Arg) and c.736T>A (p.Cys246Ser) in the LIPH gene may contribute to the clinical phenotype of the two patients with ARWH in this family.

Sequential treatment of initially diagnosed systemic multiple metastases advanced prostate cancer / 中华泌尿外科杂志

sequential treatment of advanced prostate cancer is a current research hotspot. One patient with advanced prostate cancer with multiple metastases at initial diagnosis in our institution was sequentially treated with endocrine, abiraterone, polymerase inhibitor, and enzalutamide with an overall survival of 35 months.

Genetic testing and genotype-phenotype analysis for a child with X-linked hypohidrotic ectodermal dysplasia / 中华医学遗传学杂志

OBJECTIVE@#To carry out genetic testing for a Chinese patient with X-linked hypohidrotic ectodermal dysplasia (XLHED) and explore its genotype-phenotype correlation.@*METHODS@#Clinical data of the patient was collected. Peripheral blood samples were taken from the patient, his parents and 100 unrelated healthy controls. Genetic variants were detected by using next-generation sequencing using a skin-disease panel through targeted capture and next generation sequencing. Candidate variant was verified by Sanger sequencing. All literature related to genetic testing of XLHED patients in China was searched in the database, and the genotypes and phenotypes of patients in the literature and the correlation between them were statistically analyzed.@*RESULTS@#A novel splice site variant c.655_689del was detected in the patient but not among his parents and the 100 unrelated healthy controls. So far 61 variants of the EDA gene have been identified among Chinese patients with XLHED, which suggested certain degree of genotype-phenotype correlation.@*CONCLUSION@#A novel c.655_689del variant has been identified in the EDA gene, which has expanded the spectrum of EDA gene variant and facilitated delineation of the genotype-phenotype correlation of XLHED.

Free thin sensate superficial circumflex iliac artery perforator flap based on lateral cutaneous branch of the subcostal nerve for repair of soft tissue defect in the foot and ankle / 中华创伤杂志

Objective:To investigate the efficacy of free thin sensate superficial circumflex iliac artery perforator flap based on lateral cutaneous branch of the subcostal nerve for repair of soft tissue defect in the foot and ankle.Methods:A retrospective case series study was made on clinical data of 9 patients with soft tissue defect in the foot and ankle admitted to 80th Group Military Hospital from December 2017 to December 2019. There were 8 males and 1 females, with the age of 28-63 years [(47.3±12.3)years] and the body mass index (BMI) of 16.7-27.8 kg/m 2 [(23.9±3.9)kg/m 2]. The size of soft tissue defect ranged from 10 cm×6 cm to 20 cm×12 cm. All patients were treated with the free thin sensate superficial circumflex iliac artery perforator flap based on lateral cutaneous branch of the subcostal nerve. After debridement or tumor resection, a thin sensate flap was harvested by intra-adipose tissue dissection between the superficial and deep fat layers. The size of flap ranged from 13 cm×8 cm and 13 cm×10 cm. The thickness of the defatted flap ranged from 3-6 mm. The distance from the anterosuperior iliac spine to the point where the lateral cutaneous branch of the subcostal nerve crossed the iliac crest ranged from 7.5-10.0 cm. The flap survival, complications, and reoperation were observed after operation. The sensory recovery of the flap was evaluated using Tinel sign and nine-grid method including monofilament touch perception, vibration perception, pinprick perception, temperature perception, and static two-point discrimination test. The joint range of motion, and shoewear and walking problems were recorded. At the last follow-up, the American Orthopedic Foot and Ankle Society (AOFAS) ankle-hindfoot score was used to assess the affected foot and ankle. The injury at the donor site was detected as well. Results:All patients were followed up for 6-35 months [(21.1±10.1)months]. All flaps survived without infection or tumor reoccurrence. One patient developed ulceration, then surplus skin on the reconstructed heel was resected. One patient underwent flap debulking and removal of internal fixation. One or more sensory modalities within the nine areas in each flap could be detected at postoperative 3-6 months. The monofilament touch, vibration, pinprick, and temperature perception were presented in almost all regions of each flap at postoperative 12 months. However, only one patient in one region was noted with the static two-point discrimination, in which the distance of the two points was set as 25 mm. The range of ankle motion was slightly limited in 2 patients who underwent osseoligamentous complex reconstruction. All patients were able to wear normal shoes and walk without pain. At the last follow-up, the AOFAS ankle-hindfoot score ranged from 78 to 97 points [(86.4±7.4)points], significantly improved from preoperative 10-70 points [(44.2±18.4)points] ( P<0.01). No patients complained of pain at the donor site, but the widening linear scar was noted. Conclusion:For medium-sized soft tissue defect of the foot and ankle, the free thin sensate superficial circumflex iliac artery perforator flap based on lateral cutaneous branch of the subcostal nerve can be defatted with the requirement and has advantages in defect site appearance, sensory restoration, wearing ordinary shoes, painless walking, good functional recovery, and minimal donor site morbidity.

Two cases of Darier′s disease: ATP2A2 gene mutation analysis and a family survey / 中华皮肤科杂志

Case 1, a 16-year-old male patient, presented with local verrucous hyperplasia and clustered brown follicular keratotic papules on the face, neck and bilateral axillae, some of which merged into plaques; his mother had similar medical history and clinical manifestations. Case 2, a 21-year-old male patient, presented with local verrucous hyperplasia and diffuse follicular keratotic papules on the head, face, neck, trunk, bilateral axillae and buttocks, some of which merged into patchy lesions; none of his family members had similar symptoms. Histopathological examination of the neck skin lesion of the case 2 showed epidermal hyperkeratosis with focal parakeratosis, suprabasal cleft with acantholysis, "villi" , corps ronds and grains in the spinous layer, and inflammatory cells infiltrating the superficial dermis. Genetic testing showed a missense mutation c.2300A>G in exon 15 of the ATP2A2 gene in both the case 1 and his mother, and a splice-region mutation c.2097+5G>A at the junction between exon 15 and intron 15 of the ATP2A2 gene in the case 2. Neither of these mutations were identified in the other family members of the 2 patients.

Seven cases of vitiligo caused by eyebrow tattooing / 中华皮肤科杂志

Objective:To report 7 cases of vitiligo caused by eyebrow tattooing.Methods:Seven cases of vitiligo caused by eyebrow tattooing were collected from Department of Dermatology, Henan Provincial People′s Hospital from December 2017 to May 2019, and their clinical features were retrospectively analyzed.Results:One month to 1 year after eyebrow tattooing, several eyebrows became white in the 7 patients. In the early stage, only several eyebrows became white, and the surrounding skin was normal, but white patches with unclear boundaries gradually appeared around the eyebrows in the later stage. Reflectance confocal scanning microscopy of skin lesions on the eyebrow showed depigmentation in the basal layer and around hair follicles, and highly refractive amorphous substances (colorants) in the superficial and middle dermis. The 7 patients all showed negative patch test reactions to eyebrow colorants but positive reactions to sodium dodecyl sulfate.Conclusion:No depigmentation was observed on the eyebrow skin in the early stage of vitiligo caused by eyebrow tattooing, and reflectance confocal scanning microscopy of eyebrow lesions may be beneficial in reducing its misdiagnosis.

Overexpression of miR-125 inhibits proliferation and invasion of nasopharyngeal carcinoma cells by down-regulating FGF-2 / 国际肿瘤学杂志

Objective:To explore the expression of miR-125 in nasopharyngeal carcinoma tissues and the possible regulatory mechanism of biological characteristics of tumor cells.Methods:Thirty cases of carcinoma and paracancerous tissues from patients with nasopharyngeal carcinoma admitted to Southern Hospital of Sixth People′s Hospital Affiliated to Shanghai Jiaotong University from June 2018 to June 2019 were collected. The expressions of miR-125 and fibroblast growth factor 2 (FGF-2) mRNA were detected by fluorescent quantitative PCR. Nasopharyngeal carcinoma CNE-2 cells were transfected by miR-125 mimic (miR-125 mimic group), and negative control group was set (NC group). Transwell chamber assay was used to determine cell invasion ability, scratch healing assay was used to determine cell migration ability, WST-1 assay was used to assess cell viability, flow cytometry and electron microscopy were used respectively to detect apoptosis and autophagy, dual luciferase reporter assay was used to analyze the target of miR-125, and Western blotting was used to detect the expressions of related proteins.Results:The relative expression of miR-125 mRNA in nasopharyngeal carcinoma tissues was 0.692±0.316, which was significantly lower than 1.501±0.748 in the adjacent tissues ( t=5.242, P<0.001). The relative expression of FGF-2 mRNA in nasopharyngeal carcinoma tissues was 1.317±0.552, which was significantly higher than 0.783±0.241 in the adjacent tissues ( t=7.360, P<0.001). The miR-125 mRNA expression of CNE-2 cells in the miR-125 mimic group was 4.091±0.145, which was significantly higher than 0.993±0.137 in the NC group ( t=85.062, P<0.001). The proliferative activities of CNE-2 cells in the miR-125 mimic group at 48 and 96 h after transfection were significantly lower than those in the NC group (0.891±0.214 vs. 1.295±0.245, t=6.802, P<0.001; 0.934±0.208 vs. 1.488±0.269, t=8.924, P<0.001). The number of transmembrane cells and cell migration rate of the miR-125 mimic group were 36 000±3 820 and (39.4±6.5)%, which were significantly lower than 74 000±7 500 and (102.7±10.6)% of the NC group ( t=24.728, P<0.001; t=27.883, P<0.001). The apoptosis rate of CNE-2 cells in the miR-125 mimic group was (22.5±1.4)%, which was significantly higher than that in the NC group (1.4±0.5)% ( t=77.740, P<0.001). The relative expression of the apoptotic protein Bax in the miR-125 mimic group was 0.983±0.158, which was significantly higher than that in the NC group (0.418±0.122; t=15.503, P<0.001), and the relative expression of Bcl-2 was 0.688±0.174, which was significantly lower than that of the NC group (1.013±0.109; t=8.670, P<0.001). Autophagy was observed in miR-125 overexpressing CNE-2 cells by electron microscopy, and the relative expression ratio of autophagy protein LC3Ⅱ/LC3Ⅰ in the miR-125 mimic group was 2.517±0.209, which was significantly higher than 1.238±0.135 in the NC group ( t=28.156, P<0.001). The expression of FGF-2 protein in the miR-125 mimic group was 0.504±0.118, which was significantly lower than 1.228±0.134 in the NC group ( t=22.210, P<0.001). The double luciferase report confirmed FGF-2 as the target gene of miR-125. At 12, 24, 48 and 96 h after the transfection, the cell proliferative activities of CNE-2 cells co-transfected by FGF-2 gene plasmid and miR-125 mimic were significantly higher than those of CNE-2 cells transfected by miR-125 mimic (all P<0.05), and the apoptosis rate was significantly lower than that of CNE-2 cells transfected by miR-125 mimic [(6.2±1.5)% vs. (17.6±2.4)%, t=22.062, P<0.001]. Conclusion:The expression of miR-125 is down-regulated in nasopharyngeal carcinoma tissues. Overexpression of miR-125 may inhibit the proliferation, migration and invasion of nasopharyngeal carcinoma CNE-2 cells and promote the apoptosis and autophagy of CNE-2 cells by down-regulating FGF-2 expression.

Community rehabilitation service based on "integrated general-specialty" model / 中华全科医师杂志

In order to provide quality rehabilitation care for patients in the hierarchical medical service system,a joint rehabilitation team of general practitioner and specialists from secondary and tertiary hospital was formed in Shanghai Xujiahui Community Health Service Center,and the community rehabilitation has been implemented with "integrated general-specialty" model.This article introduces the characteristics,structure,service contents and advantages of the "integrated general-specialty" rehabilitation service mode,also presents suggestions for solving the existing problems of the mode.

Mutation analysis in a pedigree with Rothmund-Thomson syndrome / 中华皮肤科杂志

Objective@#To detect gene mutations in a pedigree with Rothmund-Thomson syndrome (RTS) .@*Methods@#Clinical data were collected from two patients (an older sister and a younger brother) and their family members in a Chinese pedigree of Han nationality with RTS. Blood samples were obtained from the two patients, their unaffected older brother, their parents and 100 unrelated healthy controls. DNA was extracted, and all the exons in the encoding area of the RECQL4 gene were amplified by PCR. Gene mutations were detected by a skin-targeted next-generation sequencing panel, and verified by Sanger sequencing.@*Results@#Two heterozygous mutations were identified in the RECQL4 gene of the two patients, including a splice site mutation c.2886-1G>A and an insertion mutation c.1013_1014insC, which were inherited from the father and mother of the patients respectively. Meanwhile, neither of the two mutations was observed in 100 unrelated healthy controls or the older brother of the patients.@*Conclusion@#The splice site mutation c.2886-1G>A and the insertion mutation c.1013_1014insC in the RECQL4 gene may contribute to the clinical phenotype of the patients in this pedigree with RTS.

Improving Effects of Echinacoside on Spatial Cognitive Function in Mice under Hypobaric Hypoxia Environment and Its Mechanism Study / 中国药房

OBJECTIVE： To study the improving effects of echinacoside （ECH） on spatial cognitive function in mice under hypobaric hypoxia environment and its mechanism. METHODS： Totally 60 mice were randomly divided into blank group （normal saline）， model group （normal saline）， positive group （Ginkgo leaf extract tablet，100 mg/kg） and ECH low-dose， medium-dose and high-dose groups （50， 75， 100 mg/kg）， with 10 mice in each group. Except for blank group， other groups were cultured in hypobaric oxygen chamber to simulate hypobaric hypoxia； they were given relevant medicine intragastrically once a day， for consecutive 7 d （Placing into hypobaric oxygen chamber immediately after medication）. Using the times of horizontal and vertical activities of mice in 2 min as index， negative emotions and spatial cognitive function were evaluated. Histopathological changes of hippocampus in mice were observed by microscopy after HE staining. The levels of SOD， CAT， GSH-Px and MDA in hippocampal tissue of mice were detected. RESULTS： Compared with blank group， the times of horizontal activities， MDA level were increased significantly in model group （P＜0.05）， while the times of vertical activities， the levels of SOD， CAT and GSH-Px were decreased significantly （P＜0.05）； the pyramidal cells in the CA1 area of the hippocampal tissue were arranged loosely， and many pyramidal cells were compressed and stained deeply. Compared with model group， the times of horizontal activities and MDA level were decreased significantly in positive group and ECH high-dose group （P＜0.05）， while the times of vertical activities， the levels of SOD， CAT and GSH-Px were increased significantly （P＜0.05）； the pyramidal cells in the CA1 region of the hippocampal tissue were abundant and closely arranged， and a few of them are constricted and deeply stained. CONCLUSIONS： ECH can improve spatial cognitive impairment of mice under hypobaric hypoxia environment， the mechanism of which may be associated with up-regulation of SOD， CAT and GSH-Px， down-regulation of MDA in the hippocampal tissue.

Mutation analysis in a pedigree with Rothmund-Thomson syndrome / 中华皮肤科杂志

Objective To detect gene mutations in a pedigree with Rothmund-Thomson syndrome (RTS).Methods Clinical data were collected from two patients (an older sister and a younger brother)and their family members in a Chinese pedigree of Han nationality with RTS.Blood samples were obtained from the two patients,their unaffected older brother,their parents and 100 unrelated healthy controls.DNA was extracted,and all the exons in the encoding area of the RECQL4 gene were amplified by PCR.Gene mutations were detected by a skin-targeted next-generation sequencing panel,and verified by Sanger sequencing.Results Two heterozygous mutations were identified in the RECQL4 gene of the two patients,including a splice site mutation c.2886-1G＞A and an insertion mutation c.1013_1014insC,which were inherited from the father and mother of the patients respectively.Meanwhile,neither of the two mutations was observed in 100 unrelated healthy controls or the older brother of the patients.Conclusion The splice site mutation c.2886-1G＞A and the insertion mutation c.1013_1014insC in the RECQL4 gene may contribute to the clinical phenotype of the patients in this pedigree with RTS.