Metastatic vascular calcification and calcinosis universalis, as severe complications of parathyroid hyperfunction and hyperparathyroidism, have attracted more attention in patients with renal secondary hyperparathyroidism and primary hyperparathyroidism. But, they are of little concern in patients with long-term negative calcium balance related parathyroid hyperfunction or hyperparathyroidism caused by calcium and/or vitamin D insufficiency (CVI). CVI is common in the population. Relatively low level of serum calcium and negative calcium balance caused by long-term CVI result in parathyroid hyperfunction or hyperparathyroidism, which may cause secretion of PTH beyond the physiological level, leading to bone absorption and release of a large amount of bone calcium into the blood. It may not only cause bone loss and osteoporosis, but also form metastatic vascular calcification or calcinosis universalis presented by cardiovascular diseases and other multi-organ lesions. Early calcium deposition can gradually fade after reasonable treatment, but middle arterial calcification is not easy to fade once it occurs. Therefore, vascular calcification and calcium deposition should be actively prevented and early screened and diagnosed. The early prevention, diagnosis and treatment of parathyroid hyperfunction or hyperparathyroidism can prevent, delay, or even reverse the occurrence and development of metastatic vascular calcification and calcinosis universalis, which is significant for disease prevention and protecting the patients' health influenced by these diseases.

The lifetime prevalence of nephrolithiasis is 15% for men, with a 5-year recurrence rate of 35% to 50% after an initial event. Although it was initially recommended to limit calcium intake in these patients, a number of studies have reported association between lower total dietary calcium intake and increased risk of incident kidney stones, and that increased calcium intake might reduce the risk of incident kidney stones. We report a 35-year-old male who presented recurrent 8-years of nephrolithiasis and urine crystal with calcium intake restriction, and had no recurrence after 5 years of follow-up after intensive calcium and vitamin D supplementation.

Objective:To explore the association of DNA methylation with immune response to hepatitis B (HepB) vaccine in Han nationality children from Guangxi province.Methods:A total of 263 children aged 8-9 months who had completed HepB immunization program were recruited from three hospitals in Guangxi province by using unmatched case-control method. Children with the HepB surface antibody concentration(Anti -HBs)<100 mIU/ml was set as the case group and ≥100 mIU/ml as the control group. Multiplex PCR and heavy sulfite sequencing were used to treat the samples. Illumina platform was used for high-throughput DNA methylation sequencing of IFNG gene target regions and CpG sites. Unconditional logistic regression was used to analyze the association between cytosine-phospho-guanosine DNA methylation at 18 loci of IFNG gene and HepB immune response level. Results:There were 104 children in the case group and 159 in the control group. The median ( Q1, Q3) level of anti -HBs in two groups were 62.34 (30.06, 98.88) mIU/ml and 1 089.10 (710.35, 1 233.45) mIU/ml. The methylation levels of IFNG_1 gene 44 and 93 locus in the case group were higher than those in the control group ( P<0.05). The unconditional logistic regression model showed that the DNA methylation level of IFNG_1 gene at 44 ( OR=1.18, 95% CI: 1.03-1.35) and 93 ( OR=1.21, 95% CI: 1.07-1.38) locus was associated with the HepB response level. Conclusion:The changes of DNA methylation at locus 44 and 93 of IFNG_1 gene may be relevant factors affecting the response level of HepB in Han nationality children from Guangxi province.

Objective:To explore the association of DNA methylation with immune response to hepatitis B (HepB) vaccine in Han nationality children from Guangxi province.Methods:A total of 263 children aged 8-9 months who had completed HepB immunization program were recruited from three hospitals in Guangxi province by using unmatched case-control method. Children with the HepB surface antibody concentration(Anti -HBs)<100 mIU/ml was set as the case group and ≥100 mIU/ml as the control group. Multiplex PCR and heavy sulfite sequencing were used to treat the samples. Illumina platform was used for high-throughput DNA methylation sequencing of IFNG gene target regions and CpG sites. Unconditional logistic regression was used to analyze the association between cytosine-phospho-guanosine DNA methylation at 18 loci of IFNG gene and HepB immune response level. Results:There were 104 children in the case group and 159 in the control group. The median ( Q1, Q3) level of anti -HBs in two groups were 62.34 (30.06, 98.88) mIU/ml and 1 089.10 (710.35, 1 233.45) mIU/ml. The methylation levels of IFNG_1 gene 44 and 93 locus in the case group were higher than those in the control group ( P<0.05). The unconditional logistic regression model showed that the DNA methylation level of IFNG_1 gene at 44 ( OR=1.18, 95% CI: 1.03-1.35) and 93 ( OR=1.21, 95% CI: 1.07-1.38) locus was associated with the HepB response level. Conclusion:The changes of DNA methylation at locus 44 and 93 of IFNG_1 gene may be relevant factors affecting the response level of HepB in Han nationality children from Guangxi province.

Aberrant de novo lipid synthesis is involved in the progression and treatment resistance of many types of cancers, including lung cancer; however, targeting the lipogenetic pathways for cancer therapy remains an unmet clinical need. In this study, we tested the anticancer activity of orlistat, an FDA-approved anti-obesity drug, in human and mouse cancer cells in vitro and in vivo, and we found that orlistat, as a single agent, inhibited the proliferation and viabilities of lung cancer cells and induced ferroptosis-like cell death in vitro. Mechanistically, we found that orlistat reduced the expression of GPX4, a central ferroptosis regulator, and induced lipid peroxidation. In addition, we systemically analyzed the genome-wide gene expression changes affected by orlistat treatment using RNA-seq and identified FAF2, a molecule regulating the lipid droplet homeostasis, as a novel target of orlistat. Moreover, in a mouse xenograft model, orlistat significantly inhibited tumor growth and reduced the tumor volumes compared with vehicle control (P < 0.05). Our study showed a novel mechanism of the anticancer activity of orlistat and provided the rationale for repurposing this drug for the treatment of lung cancer and other types of cancer.
Animals ; Cell Death ; Cell Line, Tumor ; Ferroptosis ; Lung Neoplasms/drug therapy* ; Mice ; Orlistat

Objective:To establish the minimum inhibitory concentration (MIC) detection method of Yersinia pestis by determining MIC of 11 kinds of antibiotics against Yersinia pestis, to master the inhibition range of common antibiotics on Yersinia pestis, and provide baseline data for clinical treatment of plague. Methods:According to Clinical Labor Standard Institution (CLSI), the agar plate dilution method was used to determine the MIC of 11 kinds of antibiotics against 118 strains of Yersinia pestis, including ofloxacin, ciprofloxacin, trimethoprim-sulfamethoxazole, kanamycin, streptomycin, ceftriaxone, ampicillin, chloramphenicol, spectinomycin, cefuroxime and tetracycline. MIC 50 and MIC 90 (the minimum concentration of drug which could inhibit 50% and 90% of bacterial growth) were calculated. The consistency was observed by comparing the results with those of the disk diffusion method. One hundred and eighteen strains of Yersinia pestis were isolated from natural plague foci of Qinghai Province and preserved by Qinghai Institute for Endemic Disease Prevention and Control. Results:Among 118 strains of Yersinia pestis tested, no single or multiple strains of Yersinia pestis resistant to 11 kinds of antibiotics were found, which was consistent with the results of the disk diffusion method. The MIC 50 and MIC 90 of 11 kinds of antibiotics against 118 strains of Yersinia pestis were obtained. Conclusions:The MIC detection method of Yersinia pestis is successfully established. This method can be used to measure the MIC of antibiotics against Yersinia pestis in high throughput and evaluate the sensitivity of Yersinia pestis to antibiotics. It is an efficient, economical and practical experimental method.

Objective To establishment a method for detection of multiple drug resistance gene of Yersina pestis using polymerase chain reaction(PCR), to provide a guidance for treatment of plague. Methods According to National Center for Biotechnology Information (NCBI) released sequences of aminoglycoside resistant genes of streptomycin resistant,strB,strA,beta lactam antibiotics resistant genes tem,shv,and ctx-m,sulfamilamide resistant genes sul1, sul2, and sul3, a pair of primers of each gene was designed. DNAs of 282 strains isolated from plague natural foci in Qinghai Province were amplified by PCR using every pair of primers. The products were separated using gel electrophoresis, and the results were visualized through a gel imaging system. The susceptibility of 282 Yersina pestis to streptomycin, sulfamethoxazole and ceftriaxone was tested by drug sensitivity test. Results The PCR amplification results of all samples were negative,and strains with streptomycin,sulfamilamide and beta lactam antimicrobial drug resistance genes were not found. Drug sensitivity test showed that 282 strains were highly sensitive to streptomycin,sulfamethoxazole and ceftriaxone sodium.The diameter of bacteriostasis ring>19,17,21 mm, respectively. Conclusions It is a feasible method to use PCR technology to detect the multiple drug resistance genes of Yersinia pestis. Using this method to systematically monitor the resistance gene of Yersinia pestis is an efficient, economical and practical experimental method, which can provide guidance for the treatment of plague disease.

Objective In order to acquaint with the prevalence of Tibetan sheep plague in this area, we conducted a serum epidemiological investigation of Tibetan sheep plague in Qinghai Province. Methods Indirect hemagglutination assay (IHA) and colloidal gold immunochromatography (GICA) were applied to test serum samples of Tibetan sheep and whole blood samples from jugular vein of Tibetan sheep were collected in 8 Prefectures of Qinghai Province from 2013 to 2016. Results A total of 86 positive Tibetan sheep serum samples with plague F1 antibody were detected by both methods, and the positive rate was 0.68％ (86/12710), the samples collected in Xinghai County Hainan Prefecture had the highest positive rate, which was 5.20％ (27/519). The Haixi Prefecture and Yushu Prefecture were historical epidemic areas, the positive rates were 0.65％(15/2313) and 0.26％(6/2293), respectively. Hainan Prefecture, Guoluo Prefacture and Huangnan Prefecture were newly confirmed epidemic areas, the positive rates were 1.61％ (28/1741), 1.01％ (15/1481), and 1.44％(19/1316), respectively. The antibody titers were 1:20 to 1:5120, the samples collected in Maqin County Guoluo Prefecture had the highest titer, namely 1 :5120. Conclusions In Qinghai Province, Tibetan sheep plague is endemic, and there are outbreaks in some regions. So we have to enhance the Tibetan sheep plague monitoring especially in Marmot plague epidemic area.

Objective To observe the correlation between protein C activity-dependent clotting time-normalized ratio (PCAT-NR) and the related blood coagulation parameters,e.g.,fibrinogen (Fib),factor Ⅶ coagulant activity (Ⅶ:C),factor Ⅷcoagulant activity (F Ⅷ:C),antithrombin (AT),D-dimer (DD) in patients with acute cerebral infarction.Methods One hundred cases of patients who were diagnosed as acute cerebral infarction according to clinical manifestations and imaging examinations were taken as the test group and 75 healthy subjects were taken as control group.The values of Fib,FⅦ:C,FⅧ:C,AT,PCAT-NR,DD were tested and the difference between the two groups were compared.The differences of Fib,FⅦ:C,FⅧ:C,DD and AT between declined PCAT-NR group and normal PCAT-NR group in the patients with acute cerebral infarction were analyzed.The correlations of PCAT-NR with other coagulation parameters in acute cerebral infarction cases were compared.Results The values of Fib (3.38 ± 1.25) g/L,F Ⅶ:C (130.5 ± 15.9) ％,FⅧ:C (135.8 ± 43.1) ％ and DD (2.12:±:3.01) mg/L in the acute cerebral infarction group were significantly higher than those of control group,while the values of AT (83.94 ± 14.95) ％ and PCAT-NR (0.87 ± 0.23) in test group were significantly lower than those the control group (P＜0.05).The values of Fib (4.03 ± 1.25)g/L,FⅦ:C (138.2 ±6.9)％ and FⅧ:C (151.5 ± 54.9)％ of PCAT-NR declined group in the patients with acute cerebral infarction were significantly higher than those of PCAT-NR normal group (P ＜ 0.05),while the values of DD,AT were not statistically different between two groups (P ＞ 0.05).The values of PCAT-NR were significantly negatively correlated with Fib,FⅧ:C and DD in the patients with acute cerebral infarction (r =-0.484,-0.356 and-0.473,respectively (all P ＜ 0.05).There was no correlation of PCAT-NR with FⅦ:C and AT (P ＞ 0.05).Conclusion The PCAT-NR decline was associated with high coagulation state in patients with acute cerebral infarction.This decline has some correlation with high level of blood clotting factor Ⅷ and Fib.

BACKGROUND:Previous study has observed that the calf acellular dermal membrane exhibits slow repair efficiency, fast degradability speed and other shortcomings in the repair of cartilage defects. OBJECTIVE:To investigate the repair effect of the col agen type Ⅱ-modified acel ular dermal membrane on cartilage defects in rabbits. METHODS:The fetal rabbit chondrocytes were seeded onto the col agen type Ⅱ-modified acel ular dermal membrane, and the composite was then observed under scanning electron microscope at 3, 7 and 14 days. Cartilage defect models were established on the bilateral femoral condyles of 24 New Zealand white rabbits, and these model rabbits were randomly allocated to three groups. The cartilage-acellular dermal membrane and cartilage-collagen type Ⅱ-modified acellular dermal membrane were implanted into the defect regions of control and experimental groups, respectively. Those received no intervention were as blank control group. Collagen type Ⅱ immunohistochemical staining and Wakitani scoring system were performed at 6 and 12 weeks postoperatively. RESULTS AND CONCLUSION:Chondrocytes grew and adhered well in the scaffold. The Wakitani scores in the experimental group were significantly lower than those in the control and blank control groups at postoperative 6 and 12 weeks (P<0.05). At 6 and 12 weeks postoperatively, collagen type Ⅱ immunohistochemical staining was the strongest in the experimental group, with yellow and brown particles in the cytoplasm;the control group was positive for collagen type Ⅱ immunohistochemical staining, while the blank control group was negative for the staining. Our findings suggest that the collagen type Ⅱ-modified acellular dermal membrane is beneficial for the repair of cartilage defects.