AIM: To investigate the expression of TNF-?, TGF-? in patients with oral lichen planus (OLP) and normal oral mucosa (NOM). METHODS: An immunohistochemical technique was performed to detect TNF-?, TGF-?_1 and TGF-?_2 expression in 22 cases with OLP and 10 normal controls. RESULTS: In lamina propria of OLP, the expression of TNF-? and TGF-?_1 were increased, whereas TGF-?_2 did changed significantly compared with control group. TNF-? positive signal were mostly found in macrophages, lymphocytes. TGF-?_1 positive cell was present in macrophages, endothelial cells and fibrocytes. CONCLUSION: TNF-? and TGF-?_1 play an important role in the development and maintenance of OLP local inflammation.

The effects of over-expression of ANXA10 gene on proliferation and apoptosis of hepato-cellular carcinoma cell line HepG2 were elucidated. The human ANXA10 gene was subcloned into the lentiviral vector, PGC-FU, to generate the lentiviral expression vector, PGC-FU-ANXA10. The corrected ANXA10 was confirmed by endoenzyme digestion, and sequencing. Recombinant lentiviruses were produced by 293T cells following the co-transfection of PGC-FU-ANXA10 with the packaging plasmids pHelper1.0 and pHelper2.0. The resulting recombinant lentiviruses carrying ANXA10 were then used to infect human embryonic kidney epithelial cells, and lentiviral particles were produced. The ANXA10 expression in 293T cells was detected by using fluorescent microscope and Western blotting. HepG2 cells were infected, and divided into PGC-Fu-ANXA10 group, PGC-Fu group and HepG2 cell group. The changes of ANXA10 mRNA and protein expression were detected by using RT-PCR and Western blotting respectively. Flow cytometry and MTT assay were performed to examine the changes in cell apoptosis and proliferation respectively. The recombinant PGC-FU-ANXA10 vector was successfully constructed, the ANXA10 protein was detected by using Western blotting, and virus titer was 2×10(8) TU/mL. The recombinant lentiviruses were effectively infected into HepG2 cells in vitro and the infection efficiency was 70%. At 72 h after infection, the ANXA10 mRNA and protein expression levels in PGC-Fu-ANXA10 group were significantly higher than in PGC-Fu group and HepG2 cell group (P<0.05); the in vitro growth inhibition rate of HepG2 cells in PGC-Fu-ANXA10 group was 24.65%, significantly higher than that in PGC-Fu group and HepG2 cell group (P<0.05), but there was no significant difference between PGC-Fu group and HepG2 cell group; the apoptosis rate in PGC-Fu-ANXA10 group, PGC-Fu group and HepG2 cell group was (51.92±1.41)%, (19.00±1.12)% and (3.59±0.89)% respectively. The apoptosis rate in PGC-Fu-ANXA10 group was significantly higher than in PGC-Fu group and HepG2 cell group (P<0.05). The recombinant lentiviruses PGC-FU-ANXA10 were constructed successfully and infected into HepG2 cells. The overexpression of ANXA10 gene can significantly inhibit proliferation and promote apoptosis of HepG2 cells in vitro.

Objective To analyze the relationship between biochemical level and severity levels and clinical,duration of disease in patients with Parkinson Disease (PD).Methods 69 patients with PD and 69 healthy persons of similar sex and age were selected in the research.Serum uric acid and lipids levels were examined and compared.Results The serum uric acid,triglycerides,total cholesterol and low-density lipoprotein cholesterol (LDL-C) were (322.48 ± 66.18) μmol/L,(1.22 ± 0.86) mmol/L,(4.70 ± 0.92) mmol/L and (3.00 ± 0.85) mmol/L in control group,and (384.23 ± 88.28) μmol/L、(1.64 ± 0.94) mmol/L、(5.37 ± 1.31) mmol/L、(3.53 ± 1.03) mmol/L in control group.The differences are significant (t =-4.68,P =0.000;t =-2.74,P =0.007;t =-2.74,P =0.007;t =-3.49,P =0.001;t =-3.27,P =0.001).Serum UA concentration and high-density lipoprotein cholesterol (HDL-C),LDL-C were lower in patients with Parkinson's disease in duration of disease more than 3 years than those in duration of disease less than 3 years (t =3.373,P =0.001;t =2.440,P =0.017).The serum UA levels of any stages of PD patients were lower than the control group (P ＜ 0.05) according to Hoehn-Yahr staging.All lipid levels in early and middle stage PD disease patients were lower than those in control group (P ＜ 0.05).Serum UA,total cholesterol and HDL-C in female PD patients were (305.69 ± 54.25) μmol/L,(4.99 ± 0.95) mmol/L,(1.25 ± 0.27) mmol/L,and (339.76 ± 73.40) μmol/L,(4.41 ± 0.81) mmol/L,(1.06 ± 0.19) mmol/L in male patients.The difference is significant (t =2.198,P =0.031;t =-2.721,P =0.008;t =-3.266,P =0.002).Multivariate logistic regression models assessed lower uric acid concentrations is the risk of PD (OR =1.01,95％ CI 1.004 ～ 1.015,P =0.001).Conclusion Biochemical level changed differently in Parkinson disease and uric acid reduction could be a risk factor for PD.

The effects of over-expression of ANXA10 gene on proliferation and apoptosis of hepato-cellular carcinoma cell line HepG2 were elucidated. The human ANXA10 gene was subcloned into the lentiviral vector, PGC-FU, to generate the lentiviral expression vector, PGC-FU-ANXA10. The corrected ANXA10 was confirmed by endoenzyme digestion, and sequencing. Recombinant lentiviruses were produced by 293T cells following the co-transfection of PGC-FU-ANXA10 with the packaging plasmids pHelper1.0 and pHelper2.0. The resulting recombinant lentiviruses carrying ANXA10 were then used to infect human embryonic kidney epithelial cells, and lentiviral particles were produced. The ANXA10 expression in 293T cells was detected by using fluorescent microscope and Western blotting. HepG2 cells were infected, and divided into PGC-Fu-ANXA10 group, PGC-Fu group and HepG2 cell group. The changes of ANXA10 mRNA and protein expression were detected by using RT-PCR and Western blotting respectively. Flow cytometry and MTT assay were performed to examine the changes in cell apoptosis and proliferation respectively. The recombinant PGC-FU-ANXA10 vector was successfully constructed, the ANXA10 protein was detected by using Western blotting, and virus titer was 2×10(8) TU/mL. The recombinant lentiviruses were effectively infected into HepG2 cells in vitro and the infection efficiency was 70%. At 72 h after infection, the ANXA10 mRNA and protein expression levels in PGC-Fu-ANXA10 group were significantly higher than in PGC-Fu group and HepG2 cell group (P<0.05); the in vitro growth inhibition rate of HepG2 cells in PGC-Fu-ANXA10 group was 24.65%, significantly higher than that in PGC-Fu group and HepG2 cell group (P<0.05), but there was no significant difference between PGC-Fu group and HepG2 cell group; the apoptosis rate in PGC-Fu-ANXA10 group, PGC-Fu group and HepG2 cell group was (51.92±1.41)%, (19.00±1.12)% and (3.59±0.89)% respectively. The apoptosis rate in PGC-Fu-ANXA10 group was significantly higher than in PGC-Fu group and HepG2 cell group (P<0.05). The recombinant lentiviruses PGC-FU-ANXA10 were constructed successfully and infected into HepG2 cells. The overexpression of ANXA10 gene can significantly inhibit proliferation and promote apoptosis of HepG2 cells in vitro.
Annexins ; genetics ; Apoptosis ; genetics ; Carcinoma, Hepatocellular ; genetics ; Cell Line, Tumor ; Cell Proliferation ; Hep G2 Cells ; Humans ; Liver Neoplasms ; genetics

Objective To investigate the dynamic changes ofprocalcitonin (PCT) within 72 h of acute stroke without infection and explore the value of PCT in diagnosis of bacterial infection in the early stage of acute stroke.Methods Forty-one patients with acute stroke within 24 hours of symptom onset,admitted to our hospital from July 2012 to January 2013,were enrolled in our study.The concentrations of PCT and C-reactive protein (CRP) in the serum were measured,respectively,at 24,48 and 72 h after symptom onset.At each time point,the PCT and CRP values were compared with the upper value of normal ranges of PCT and CRP,respectively.Results The median (quartiles) PCT concentrations at 24,48 and 72 h after stroke onset were,respectively,(0.050 [0.040,0.080]) ng/mL,(0.060 [0.036,0.095]) ng/mL and [0.051 (0.040,0.079)] ng/mL,which were significantly different as compared with that of the upper value of normal range (0.05 ng/mL,P＜0.05).The median (quartiles) CRP concentrations at 24 and 48 h after stroke onset were,respectively,[3.200 (1.100,5.000)] mg/L and [4.300(1.700,9.900)] mg/L,showing no significant difference with the upper value of normal range (5.0 mg/L,P＞0.05); however,the mean CRP concentration at 72 after stroke onset was [5.300 (2.500,15.550) mg/L],enjoying significant difference as compared with the upper value of normal range (P＜0.05).Most of the patients (22 patients,53.67％) had a peak level of PCT at 24 h,while most of them (26,63.41％) had a peak level of CRP at 72 h.The concentration of PCT increased within 24 h after symptom onset,but declined in the following 72 h; in contrast,the concentration of CRP continuously increased in the first 72 h of symptom onset.Conclusions PCT concentrations may increase in the first 72 h after acute stroke,therefore,when using PCT in diagnosis of bacterial infection in the early stage of acute stroke,the influence of elevating PCT concentrations by stroke itself should be considered.But PCT usually reaches its peak level earlier than CRP and returns to normal range faster than CRP,which may be more valuable than CRP in diagnosis of bacterial infection in the early stage of acute stroke.

Objective:To analyze changes in energy metabolism and oxylipin metabolism in macrophages after stimulation by Mycobacterium marinum ( M. marinum) using targeted metabolomics, and to provide insights into the mechanisms underlying the immune defense by macrophages against M. marinum infections. Methods:Mouse bone marrow-derived macrophages were obtained from the bilateral femurs of mice, and cultured cells were divided into two groups: the active M. marinum group and the inactivated M. marinum group. Bacterial suspensions were prepared using M. marinum clinical isolates; the active M. marinum group was treated with live M. marinum suspensions for 12 hours, while the inactivated M. marinum group with inactivated M. marinum suspensions for 12 hours. Cell morphology was observed through microscopy, and cell length was measured. Cell lysates collected from both groups were subjected to liquid chromatography-tandem mass spectrometry analysis to detect energy and oxylipin metabolites. A t-test was utilized to compare the lengths of macrophages between the two groups, while principal component analysis and orthogonal partial least squares-discriminant analysis were conducted to identify differential metabolites. Results:Under the microscope, macrophages in the active M. marinum group formed more granuloma-like cell aggregates compared with those in the inactivated M. marinum group; the macrophages were significantly thinner and longer in the inactivated M. marinum group (439.52 ± 91.67 μm) than in the active M. marinum group (289.96 ± 70.11 μm, P < 0.001). Principal component analysis and orthogonal partial least squares-discriminant analysis of energy metabolism and oxylipin metabolism in macrophages demonstrated good separation between the two groups. As for the energy metabolism, a total of 12 differential metabolites were identified, with the amino acid metabolism showing the most significant changes. Specifically, there was a significant increase in the content of L-citrulline, while the content of L-leucine and serine decreased. As for the oxylipin metabolism, 20 differential metabolites were identified, with the arachidonic acid metabolism showing the most significant changes. Conclusions:Macrophages stimulated by live M. marinum exhibited altered amino acid metabolism and arachidonic acid metabolism compared with those stimulated by inactivated M. marinum, characterized by an increase in L-citrulline content, a decrease in L-leucine and serine levels, and alterations in arachidonic acid content.

Objective:To explore the functional connection (FC) changes of default mode network (DMN) with the patients who have clinically remitted from major depressed disorder (MDD) and the brain imaging basis of the rehabilitation mechanism of major depression.Methods:Seventeen right-handed outpatients in the medical and psychological ward of Nanjing Brain Hospital who met the inclusion criteria were recruited by psychiatrists.MDD subjects after enrollment were scanned by resting-state functional magnetic resonance (fMRI) in baseline period(rMDD) and a 6-month follow-up period (sMDD). Tweenty-two healthy controls (control group, HCs group) matched with the MDD group in gender, age, and educational level were recruited and given the same resting fMRI scan.The independent component analysis (ICA) was used to extract DMN brain regions of rMDD, sMDD and HCs in resting state separately and to compare the changes of DMN functional connectivity in full remitted MDD patients.Results:The DMN data showed that the functional connectivity of right orbital middle frontal gyrus(MNI: x, y, z=3, 54, -9), right posterior cingulate(MNI: x, y, z=6, -51, 30), and precuneus(MNI: x, y, z=9, -54, 27) in the rMDD was significantly higher than the HCs group, and no functional connectivity in the rMDD was found to be lower than that in HCs group.Furthermore, no significant difference been found between the sMDD and HCs group.Compared with the rMDD, the functional connectivity of orbital middle frontal gyrus(MNI: x, y, z=3, 54, -9) and left medial prefrontal cortex (mPFC) (MNI: x, y, z=-3, 66, 12)in the sMDD was significantly lower than that in the rMDD, and the functional connectivity of left angular gyrus in the sMDD(MNI: x, y, z=-57, -57, 33) was significantly higher than that in the rMDD.Conclusion:The DMN network has not fully returned to its normal level though the posttreatment Hamilton Depression Scale-17 score was lower than 7 points, and the injury of FC is still recovering in the following 6 months, suggesting that the recovery of the DMN network function was delayed in the full remission of clinical symptoms, and is related to the recure of MDD.

This study reported two women with extreme obesity who underwent metabolic surgery due to their mutations in leptin receptor (LEPR).Genomic DNA was extracted from the anticoagulant blood samples of the two patients and their parents.A panel of genes related to metabolic diseases or whole exon sequencing was screened and the results were confirmed by Sanger sequencing.This is the first time that these three mutations in LEPR were reported.Two patients complained insatiety and early-onset obesity since childhood at clinics.Patient 1 was a 39-year-old woman with height 150 cm,weight 130 kg,and BMI 57.8 kg/m2.Serum leptin level was 156.4 μg/L.A homozygous mutation of c.2317G＞T was found in exon 15 of LEPR gene in patient 1,which was descended from her father and mother respectively.Patient 2 was a 37-year-old woman with height 158 cm,weight 167 kg,and BMI 67 kg/m2.Serum leptin level was 193.4 μg/L.Genetic analysis showed compound heterozygous mutations of c.1482delT and c.1892C ＞ A.Her father showed heterozygous c.1482delT mutation,and her mother carried heterozygous c.1892C ＞ A mutation.Two patients all underwent metabolic surgery with body weight reduction of about 22 kg and 40 kg respectively after first six months.However,the follow-up studies showed that the body weight of patient 1 rebounded to pre-surgery level in two years and patient 2 did not further lose weight in the following six months.

Objective To explore the value of pulse index continuous cardiac output (PICCO) combined with intracranial pressure monitoring in patients with severe craniocerebral injury.Methods One hundred and thirty-eight patients with severe craniocerebral injury accepted controlling decompression surgical treatment in our hospital from February 2015 to February 2019 were prospectively chosen.According to patients' families will,postoperative application of PICCO combined with intracranial pressure monitoring for fluid management was performed in 72 patients (treatment group) and application of central venous pressure combined with intracranial pressure monitoring for fluid management was performed in 66 patients (control group).All patients were adjusted according to the monitoring results.The intracranial pressure and cerebral perfusion pressure one week after surgery,incidences of new traumatic cerebral infarction,neurogenic pulmonary edema,pulmonary infection,scalp exudation,and intracranial infection,average hospitalization days,total hospitalization costs,intensity of antimicrobial use,and Glasgow coma scale scores two weeks after operation were compared and analyzed between the two groups.Glasgow outcome scale was used to evaluate the prognoses of the patients 6 months after injury.Results There were 7 patients (3 from the control group and 4 from the treatment group) dropped out of the study due to various reasons and 131 patients (63 from the control group and 68 from the treatment group) included in the final statistical analysis;there was no significant difference in drop-out rate of the two groups (P＞0.05).The intracranial pressure in the treatment group ([14.28±2.98] mmHg) was significantly lower than that in the control group ([18.99±2.78] mmHg) and cerebral perfision pressure ([66.72±2.25] mmHg) was significantly higher than that in the control group ([52.96±3.12] mmHg) one week after operation (P＜0.05).During hospitalization,the incidences of new traumatic cerebral infarction,neurogenic pulmonary edema,pulmonary infection,scalp exudation and intracranial infection in the treatment group (8.8％,13.2％,11.8％,7.4％,and 2.9％) were significantly lower than those in the control group (22.2％,27.0％,25.4％,19.0％,and 12.7％,P＜0.05).The average hospitalization days,total hospitalization expenses and intensity of antimicrobial use in the treatment group were significantly shorter/lower than those in the control group (P＜0.05).Glasgow coma scale scores (11.88±1.78) and good recovery rate (76.5％) in the treatment group were significantly higher than those in the control group (8.06±1.12,54.0％) two weeks after operation (P＜0.05).Good recovery rate (76.5％) in the treatment group was significantly higher than that in the control group (54.0％,P＜0.05).The mortality rate (5.9％) was significantly lower than that in the control group (17.5％,P＜0.05).Conclusion PICCO combined with intracranial pressure monitoring can effectively improve intracranial pressure,optimize cerebral perfusion,reduce complications such as traumatic cerebral infarction,neurogenic pulmonary edema,pulmonary infection and intracranial infection in patients with severe craniocerebral injury,thereby improving prognosis and reducing mortality;besides that,it can reduce patients' exposure to anti-brain infection,and the breadth and intensity of bacterial drugs can reduce the length of hospitalization and total cost of hospitalization,thereby reducing the burden of family and society.

Objective:Non-alcoholic fatty liver disease(NAFLD)is a common metabolic disorder in overweight and obese children,and its etiology and pathogenesis remain unclear,lacking effective preventive and therapeutic measures.This study aims to explore the association between whole blood copper,zinc,calcium,magnesium and iron levels and NAFLD in overweight and obese children aged 6 to 17 years,providing a scientific basis for the prevention and intervention of early NAFLD in overweight and obese children. Methods:A cross-sectional study design was used to collect relevant data from overweight and obese children who visited the Hunan Children's Hospital from January 2019 to December 2021 through questionnaire surveys.Fasting blood samples were collected from the subjects,and various indicators such as blood glucose,blood lipid,and mineral elements were detected.All children were divided into an overweight group(n=400)and a NAFLD group(n=202).The NAFLD group was divided into 2 subgroups according to the ALT level:A non-alcoholic fatty liver(NAFL)group and a non-alcoholic steatohepatitis(NASH)group.Logistic regression analysis was used to analyze the association between minerals(copper,zinc,calcium,magnesium,and iron)and NAFLD,NAFL and NASH. Results:A total of 602 subjects were included,of whom 73.6%were male,with a median age of 10(9,11)years,and a body mass index(BMI)of 24.9(22.7,27.4)kg/m2.The intergroup comparison results showed that compared with the overweight group,the NAFLD group had higher levels of age,BMI,diastolic blood pressure(DBP),systolic blood pressure(SBP),triglyceride(TG),low density lipoprotein(LDL),alanine transaminase(ALT)and aspartate aminotransferase(AST),and lower level of high density lipoprotein(HDL).The NAFL group had higher levels of age,BMI,DBP,SBP,ALT,and AST,and lower levels of HDL compared with the overweight group.The levels of age,BMI,DBP,SBP,TG,LDL,ALT,and AST of NASH were higher than those in the overweight group,while the level of HDL was lower than that in overweight group(all P<0.017).After adjusting for a variety of confounders,the OR of NAFLD for the highest quantile of iron was 1.79(95%CI 1.07 to 3.00)compared to the lowest quantile,and no significant association was observed between copper,zinc,calcium,and magnesium,and NAFLD.The subgroup analysis of NAFLD showed that the OR for the highest quantile of iron in children with NAFL was 2.21(95%CI 1.26 to 3.88),while no significant association was observed between iron level and NASH.In addition,no significant associations were observed between copper,zinc,calcium,and magnesium levels and NAFL or NASH. Conclusion:High iron level increases the risk of NAFLD(more likely NAFL)in overweight and obese children,while copper,zinc,calcium,magnesium,and other elements are not associated with the risk of NAFLD in overweight and obese children.