Objective In China primary hyperparathyroidism is not a kind of common disease as in the wesyrn countries.This article reports the current status in the diagnosis and treatment of primary hyperparathyroidism in the mainland of China. Methods We collected 730 cages of primary hyperparathyroidism diagnosed and treated in 7 top hospitals for endocrine surgery from 1965 to 2005.Results In this study.652(89.3%)cases were clinically symptomatic while 78(10.7%)cases were asymptomatic:442 cases were positive on 99mTc-MIBI scanning.Bilateral explorations were undertaken in 377 patients and unilateral or uni-gland exploration through the conventional incision in 204 cases.Minimally invasive parathyroidectomy in 143 cases.Endoscopically assisted 2 cm incision was taken in 6 cases for unilateral gland exploration.Pathologically 632(86.6%)cases were identified as adenoma,58(8.3%)cases were of hyperplasia and 40(5.5%)cases were of carcinoma.There were no major postoperative complications.While 20 patients suffering from recurrence or persistent postoperative hyperparathyroidism,the others are of normal or depressed serum level of calcium. Conclusions Preoperative localization is very helpful: Unilateral exploration for parathyroid adenoma is feasible; minimally invasive parathyroidectomy throush minimal incision is a kind of improving procedure for the localized parathyroid adenoma.

Objective·To study the effect of phosphatase and tensin homolog on chromosome 10(PTEN)on alternative splicing of forkhead box Ml(FoxM1),and its impact on tumor cell migration.Methods·PTEN was knocked down by short hairpin RNA(shRNA)in human embryonic kidney 293T cells,human prostate cancer DU145 cells,human colorectal adenocarcinoma RKO cells,and human colon cancer SW480 and SW620 cells.Specific primers were designed for FoxM1 and its subtypes FoxM1B and FoxM1C,and the mRNA expression levels of FoxM1B and FoxM1C were detected by quantitative real-time PCR(qRT-PCR).FoxM1B and FoxM1C were overexpressed in DU 145 cells,and their effects on tumor cell migration were tested by Transwell assay and wound healing assay.Immunofluorescence and dual luciferase reporter gene assay were used to explore the potential mechanism of differential regulation of tumor cell migration by FoxM1B and FoxM1C.Results·① PTEN was knocked down in 293T,DU 145,RKO,SW480,and SW620 cell lines.qRT-PCR results showed that compared with the control cells,the mRNA expression level of FoxM1B significantly increased in PTEN knockdown cells,while the mRNA expression level of FoxM1C decreased or remained unchanged.Knockdown of PTEN did not affect the transcription level of FoxM1,but caused the variable splicing of FoxM1 and promoted the generation of FoxM1B.② Compared with the control cells,the number of DU 145 cells migrating to the below chamber increased in the FoxM1B overexpression group(P=0.024),while the number of migrating DU 145 cells in the FoxM1C overexpression group was lower(P=0.000).The healing ability of DU 145 cells was significantly enhanced in the FoxM1B overexpression group(P=0.001),while the healing ability of DU145 cells was weakened in the FoxM1C overexpression group(P=0.021).Overall,FoxM1B and FoxM1C had opposite effects on tumor cell migration.FoxM1B promoted tumor cell migration,while FoxM1C inhibited tumor cell migration.③ Neither FoxM1B nor FoxM1C overexpression could induce β-catenin to enter the nucleus.Dual luciferase reporter gene assay showed no difference in the transcriptional activity of FoxM1B and FoxM1C.The difference between FoxM1B and FoxM1C in the regulation of tumor metastasis was also not mediated by β-catenin translocation.Conclusion·Knockdown of PTEN regulates the alternative splicing of FoxM1,leading to increasing expression of transcript FoxM 1B,which plays a positive role in tumor cell migration.

Objective·To investigate the biological roles and associated mechanisms of the hypoxia-induced long non-coding RNA 68(HILRNA68)in hepatocellular carcinoma(HCC)cell lines.Methods·Long non-coding RNA(lncRNA)microarray analysis was conducted to study the differential expression of lncRNAs in the HCC cell lines cultured under hypoxia treatment and normoxia treatment separately for 12 h,and DEseq2 R package was used for the analysis of differentially expressed lncRNAs.Quantitative real-time polymerase chain reaction(qRT-PCR)was used to determine the differential lncRNAs.Short hairpin RNAs(shRNAs)were used to knock down hypoxia-inducible factors(HIFs)to investigate whether HILRNA68 transcription was regulated by HIFs under hypoxia.Nucleus-cytoplasmic isolation combined with qRT-PCR and RNA fluorescence in situ hybridization(RNA-FISH)experiments were used to investigate the subcellular localization of HILRNA68.HILRNA68 was knocked down in SMMC-7721 and MHCC-97H cells by small interfering RNA(siRNA)to investigate its cellular function under hypoxia.The impact of HILRNA68 on the cell proliferation and invasion capabilities of HCC cells under hypoxia was examined by cell counting and Transwell assays.Dual-luciferase reporter assay was employed to identify how HILRNA68 regulated the transcriptional activity of HIFs under hypoxia.Results·By differential expression analysis of lncRNAs,a total of 247 and 17 significantly(defined as fold change≥4,FDR≤0.05)up-and down-regulated IncRNAs,respectively,were identified.Among these differentially expressed genes,IncRNA HILRNA68 was up-regulated about 10-fold in multiple HCC cell lines when cultured under hypoxia for 12 h.Knockdown of HIF1α,HIF2α,and HIF1β significantly suppressed(all P＜0.05)the upregulation of HILRNA68 under hypoxia.Luciferase reporter assay suggested that the transcription of HILRNA68 was regulated by HIFs.Subcellular localization studies revealed that HILRNA68 was mainly localized in the nucleus.Biological function experiments showed that silencing of HILRNA68 significantly inhibited the proliferation and invasion of HCC cells under hypoxia(all P＜0.05).Mechanistic studies demonstrated that knock-down of HILRNA68 significantly suppressed the transcriptional activity of HIF1α under hypoxia(P＜0.05)and the up-regulation of these canonical HIFs targets under hypoxia was also significantly inhibited after HILRNA68 knockdown(P＜0.05).Conclusion·The current study identifies a series of differential hypoxia-regulated lncRNAs and functionally annotates the upregulated HILRNA68.HILRNA68 is directly up-regulated by HIFs which promotes cell proliferation and invasion under hypoxia.Mechanistically,the upregulation of HILRNA68 under hypoxia enhances the transcriptional activity of HIF1α.

AIM: To investigate the implication of respiratory microbiology on the efficacy of PD-1 inhibitors monotherapy for patients with NSCLC. METHODS: This study was designed as a retrospective study, fifty-eight patients with previously-treated advanced NSCLC who were received PD-1 monotherapy from October 2018 to October 2021 were included. The PD-1 inhibitors were consisted of camrelizumab, sintilimab and pembrolizumab. Additionally, the basic demographic data, therapeutic efficacy data, survival prognosis and adverse reactions during the PD-1 inhibitors treatment were collected and analyzed through the patients' medical records of the department and the electronic medical record system of the hospital. Furthermore, deep induced sputum specimens of the patients before treatment with PD-1 inhibitor were collected. And the respiratory microbiology of 58 samples were detected using 16S rRNA gene sequencing method. The index of respiratory microbiology α diversity was analyzed, and the correlation analysis was performed with the efficacy and prognosis of patients. RESULTS: A total of 58 patients with advanced NSCLC met the study's screening criteria and were evaluable for efficacy and safety profile. Efficacy data suggested that the objective response rate (ORR) and disease control rate (DCR) of the patients who received PD-1 inhibitors was 19.0%(95% CI: 9.9%-31.4%) and 55.2% (95% CI: 41.5%-68.3%). Furthermore, prognostic data obtained from follow-up indicated that the median PFS of the 58 patients with advanced NSCLC was 3.2 months (95% CI: 2.29-4.11) and the median OS was 10.5 months (95% CI: 5.58-15.43). Regarding the exploratory analysis between efficacy and respiratory microbiology, the 58 patients with NSCLC were divided into high α diversity group (group H) and low α diversity group (group L) according to Shannon diversity index of respiratory microecology detection. And the association analysis suggested that the ORR of patients with group H and group L was 23.3% and 17.9%(P c 0.380), respectively. Furthermore, prognostic analysis indicated that the median PFS of patients with group H and group L was 3.8 and 2.8 months, respectively, which was statistically significant (P c 0.034). CONCLUSION: PD-1 inhibitors monotherapy demonstrated preliminary efficacy and prognosis as subsequent line treatment for patients with advanced NSCLC. Patients with higher α -diversity of respiratory microbiology might confer a superior prognosis. And the conclusion should be validated in large sample prospective clinical trials subsequently.