Objective To investigate the clinical characteristics,treatment effect,long-term follow up results,guanosine triphosphate (GTP) cyrclohydrolase Ⅰ (GCH Ⅰ)gene and tyrosine hydroxylase(TH) gene mutations in patients with dopa-responsive dystonia (DRD).Methods The clinical features of 3 families with 4 affected members were analyzed and all of 4 patients were screened for mutations of the GCH Ⅰ gene and TH gene with DNA sequences.Results Four patients were females,average age at onset was (15.3 ± 5.6) years (range:from 9 to 20 years).The initial symptoms were a gait disorder,stiffness or tremor of the lower limbs in all patients presented with diurnal fluctuation.As the increase of disease duration,bilateral hand tremor was found in three patients,systemic torsion was found in one patient and torticollis was found in one patient.All patients' symptoms were in complete remission after administration of low dose of levodopa.Four patients were followed up for 0.5 to 10.0 years,and all were still responsive to the levodopa treatment and effective dosage was decreased as the increase of the disease duration.No longterm side effects of levodopa had occurred after long-term treatment.One patient was found to have c.607G ＞A(p,Gly203Arg) heterogenetic mutation in GCH I gene.Molecular analysis revealed a compound heterozygous mutation in the TH gene (p.Y447Ter and p.V468M) in one patient.No point mutations in both genes were found in other patients.Conclusions DRD patients have dramatic and sustained response to levodopa and no long-term side effects of levodopa after long-term treatment.The detection of GCH Ⅰ and TH gene mutations is helpful in early diagnosis but the negative results could not exclude the diagnosis of DRD.

Objective To explore the surgical treatment of Crohn's disease(CD).Methods Clinical data of 11 patients with Crohn's disease undergoing surgery were retrospectively analyzed.Results 9 cases were diagnosed before operation,with symptoms including abdomen pain,diarrhea or constipation,weight loss,and segmental lesions.Abdominal mass was the most common cause,accounting for 54.5%(6/11)in surgery,and intestinal obstruction was secondary.accounting for 36.4%(4/11),and perianal abscess,9.1%(1/11).Partial enterectomy and anastomosis was the main procedure.3 cases were suspected malignance and underging radical cure.The pathology results showed there was moderate atypical hyperplasia in 2 of 3.Most of the patients had a good recovery and their nutritional conditions were improved obviously(P＜0.05).Conclusion Abdominal mass and intestinal obstruction are the main causes of surgical management in patients with Crohn's disease.The possibility of cancerization is higher in patients with longer medical history.The length of intestine reseeted would be enough with visitable lesions resected,and the operative effects are as good as those underwent radical cure.

We reported a case of polyglandular syndrome induced by programmed death-1(PD-1) inhibitors. The patient was a 51-years-old male with non-small cell lung cancer, treated with PD-1 inhibitor nivolumab/pembrolizumab because of postoperative subcarinal lymph node metastasis indicated by PET-CT. During 14 cycles of PD-1 inhibitor treatment, the patient successively developed primary hypothyroidism, and type 1 diabetes mellitus(T1DM). More than five months after the withdrawal of pembrolizumab, the patient experienced recurrentce. Laboratory examinations showed mild hyponatremia and hypopituitarism including ACTH and growth hormone(GH)/insulin-like growth factor-1(IGF-1) insufficiency. This is the first report of a patient diagnosed as polyglandular syndrome caused by PD-1 inhibitor. In particularly, the hypothyroidism and T1DM did not improve after drug withdrawal, while hypopituitarism was further aggravated. This case reminds us that we should pay more attention to the changes of endocrine function during and after the treatment of PD-1 inhibitor, so that we can make the correct diagnosis and take proper medical measures timely, to avoide missed diagnosis, and improper treatment.

Objective:To investigate the mechanism of oxidative stress injury and possible pathways in glutaryl CoA dehydrogenase deficient (GCDH -/-) rats with high lysine diet (Lys). Methods:Four-week-old rats were randomly divided into 6 groups: wild type+standard diet group (WT, n=6), GCDH -/-+standard diet group (GCDH -/-, n=11), WT+Lys group ( n=8), GCDH -/-+Lys group ( n=13), WT+Lys+vitamin (V) E group ( n=7), and GCDH -/-+Lys+VE group ( n=12); rats in the WT group and GCDH -/- group were given standard diet, and rats in the WT+Lys group, GCDH -/-+Lys group, WT+Lys+VE group and GCDH -/-+Lys+VE group were given high lysine diet (4.7% Lys); rats in the WT+Lys+VE and GCDH -/-+Lys+VE group were given VE (100 mg/[kg·d]) by intragastric administration once per d, and rats in other groups were given normal saline by intragastric administration once per d. The body mass and survival of rats in each group were observed. Twenty-eight d after intervention, rats were injected intraperitoneally with 10% chloral hydrate and anesthetized; their brains were severed to obtain hippocampal tissues; and pathomorphological changes were observed by HE staining; the content/activity of glutathione peroxidase (GPx), malondialdehyde (MDA), superoxide dismutase (SOD), catalase (CAT) and reduced glutathione (GSH) in the hippocampus were detected by ELISA; the protein expressions of P38, c-Jun N-terminal kinase (JNK) and extra-celluar regulated protein kinase (ERK) in the hippocampus were detected by Western blotting. Results:(1) The survival ratio of rats in the GCDH -/-+Lys group was 9/13, and that in the GCDH -/-+Lys+VE group was 11/12. From the 7 th d of intervention, the body mass of rats in the GCDH -/-+Lys group and GCDH -/-+Lys+VE group was significantly lower than that in the WT group ( P<0.05). (2) As compared with that in the WT group, MDA content in hippocampal tissues of rats in the GCDH -/-+Lys+VE group and GCDH -/-+Lys+VE group was significantly increased ( P<0.05). As compared with WT group, GCDH -/-+Lys group had significantly decreased GPx activity, CAT activity and SOD activity, and statistically decreased GSH content ( P<0.05). As compared with those in the GCDH -/-+Lys+VE group, the GPx activity, CAT activity, SOD activity, and GSH content in the GCDH -/-+Lys+VE group were significantly increased ( P<0.05). (3) Western blotting showed that as compared with that in the WT group, the P38 protein expression in the hippocampus of rats in GCDH -/-+Lys group and GCDH -/-+Lys+VE group was significantly increased ( P<0.05); as compared with GCDH -/-+Lys+VE group, the P38 protein expression in the GCDH -/-+Lys+VE group was statistically decreased ( P<0.05). Conclusion:There is oxidative stress injury in the hippocampus of GCDH -/- rats with Lys, whose possible mechanism is to activate P38 and initiate MAPK signaling pathway; VE protects GCDH -/- hippocampal cells from oxidative stress by decreasing P38 expression.

Accurate collection and preservation of vitreous and retina-related tissue specimens is the basis for clinical diagnosis and rigorous basic research. The clinical uses of vitreous specimens include microbial culture, cytological detection, detection of degenerative diseases, PCR analysis, and cytological detection of cell morphology. The experimental research uses include DNA gene analysis, protein quantitative analysis, metabolite examination, RNA content quantitative analysis, cytokine determination and so on. Retinal specimens collecting was mainly used for PCR analysis of retinal proliferative membrane, immunohistochemical staining, immunofluorescence examination, microvascular density evaluation, cell isolation and culture, etc. Understanding the collection of vitreoretinal surgical specimens and the application of relevant detection techniques and materials can provide a more comprehensive idea for the diagnosis of vitreoretinal diseases and a broader reference for the related basic research.

Objective To predict and verify the upstream regulatory microRNA (miRNA)of protein kinase D1 (PKD1),and to investigate its role in cerulein induced acute pancreatitis (AP)in rats. Methods Potential up-stream regulatory miRNA of PKD1 was predicted by using bioinformatics software. Dual luciferase reporter gene system and Western blot were applied to verify the regulation of PKD1 by the selected miRNA. Experimental AP was induced by 6 intraperitoneal injection of cerulein (20 μg/ kg)at hourly intervals after administration of the CY5 - labeled notar-get control (AP group,n = 20)or selected miRNA (treatment group,n = 20),respectively by intraperitoneal injection into rats. Other rats were divided randomly into a normal control group (n = 10)without any treatment. Besides 10 rats in either AP or treatment group were sacrificed 6 hours after the first injection of cerulein,and the rats were all sacri-ficed 24 hours after the first injection. The blood samples and pancreatic tissues of each rat were collected to test serum amylase and lipase activities,or to make hematoxylin - eosin stain for AP pathological scores as well as PKD1 immuno-histochemical staining,respectively. Results TargetScan 7. 1 software analysis showed that miR - 128 - 3p was the po-tential upstream regulatory miRNA of PKD1,which was verified by dual luciferase reporter gene system and Western blot detection. Compared to the normal control group,serum amylase and lipase activities after 6 h exposure to cerulein increased in both AP group and the treatment group[13313. 00(9424. 00 - 15995. 00)U/ L,13552. 00(10399. 50 -18408. 25)U/ L vs. 1430. 50(1214. 25 - 1543. 25)U/ L;547. 00 (515. 00 - 627. 00)U/ L,857. 50(522. 00 -1222. 25)U/ L vs. 34. 00(32. 50 - 34. 75)U/ L],and the differences were significant(χ2 = 8. 715,P < 0. 05;χ2 =9. 115,P < 0. 05),which indicated that the rat models of AP were successfully established. The immunohistochemical scores of PKD1 after 24 h exposure to cerulein decreased in the treatment group[0. 50(0 - 2. 75)scores],compared with the normal control group [4. 00(4. 00 - 8. 00)scores]and the AP group [4. 00(3. 75 - 8. 00)scores],and difference was significant(χ2 = 18. 302,P < 0. 05). Accordingly,the total pathological scores of HE staining decreased significantly in the treatment group,as compared to the AP group (3. 80 ± 0. 85 vs. 6. 90 ± 1. 15,t = 4. 481,P < 0. 01). The results showed that the inflammatory cell infiltration and tissue necrosis were significantly improved after miR -128 - 3p treatment. Conclusions miR - 128 - 3p is the upstream regulatory microRNA of PKD1 which protects pan-creata from necrotic injury and inflammatory cell infiltration in PKD1 - mediated acute pancreatitis.