[Effects and molecular mechanism of exogenous L-carnitine on excessive endoplasmic reticulum stress-mediated hepatic pyroptosis in severely scald rats].

Objective: To investigate the effects and molecular mechanism of exogenous L-carnitine on hepatic pyroptosis mediated by excessive endoplasmic reticulum stress in severely scald rats. Methods: The experimental research method was adopted. According to the random number table (the same group method below), fifteen female Sprague Dawley rats aged 6-8 weeks were divided into sham-injury group, scald alone group, and scald+carnitine group (with 5 rats in each group), and full-thickness scald of 30% total body surface area were made on the back of rats in scald alone group and scald+carnitine group, and rats in scald+carnitine group were additionally given intraperitoneal injection of L-carnitine. At post injury hour (PIH) 72, The levels of aspartate aminotransferase (AST) and alanine dehydrogenase (ALT) of biochemical indicators of liver injury were detected by automatic biochemical analyzer with the sample number of 5. At PIH 72, liver tissue damage was detected by hematoxylin-eosin staining. At PIH 72, The mRNA levels of nucleotide-binding oligomerization domain-containing protein-like receptor family pyrin domain containing 3 (NLRP3), cysteine aspartic acid specific protease 1 (caspase-1), gasderminD (GSDMD), and interleukin 1ß(IL-1ß) in liver tissue as pyroptosis-related markers and glucose regulatory protein 78 (GRP78) and CCAAT/enhancer binding protein homologous protein (CHOP) in liver tissue as endoplasmic reticulum stress-related markers were detected by real-time fluorescence quantitative reverse transcription polymerase chain reaction (RT-qPCR). Protein expression levels of GRP78, CHOP, NLRP3, caspase-1, caspase-1/p20, GSDMD-N, and cleaved IL-1ß in liver tissue were detected by Western blotting, and the sample numbers were all 5. HepG2 cells as human liver cancer cells were divided into dimethyl sulfoxide (DMSO) group, 0.1 µmol/L tunicamycin (TM) group, 0.2 µmol/L TM group, 0.4 µmol/L TM group, and 0.8 µmol/L TM group and were treated accordingly. After 24 h of culture, cell viability was detected by cell counting kit 8, and the intervention concentration of TM was screened, and the sample number was 5. HepG2 cells were divided into DMSO group, TM alone group, and TM+carnitine group, and treated accordingly. After 24 h of culture, the protein expression levels of GRP78, CHOP, NLRP3, caspase-1, caspase-1/p20, GSDMD-N, and cleaved IL-1ß in cells were detected by Western blotting, and the sample numbers were all 3. Data were statistically analyzed with one-way analysis of variance and least significant difference-t test. Results: At PIH 72, the AST and ALT levels of serum in scald alone group were (640±22) and (157±8) U/L, which were significantly higher than (106±13) and (42±6) U/L in sham-injury group, respectively, with t values of -46.78 and -25.98, respectively, P<0.01. The AST and ALT levels of serum in scald+carnitine group were (519±50) and (121±10) U/L, which were significantly lower than those in scald alone group, respectively, with t values of 4.93 and 6.06, respectively, P<0.01. At PIH 72, the morphology of liver tissue of rats in sham-injury group were basically normal with no obvious inflammatory cell infiltration; compared with those in sham-injury group, the liver tissue of rats in scald alone group showed a large number of inflammatory cell infiltration and disturbed cell arrangement; compared with that in scald alone group, the liver tissue of rats in scald+carnitine group showed a small amount of inflammatory cell infiltration. At PIH 72, the mRNA expression on levels of NLRP3, caspase-1, GSDMD, and IL-1ß in liver tissue of rats in scald alone group were significantly higher than those in sham-injury group (with t values of 34.42, 41.93, 30.17, and 15.68, respectively, P<0.01); the mRNA levels of NLRP3, caspase-1, GSDMD, and IL-1ß in liver tissue of rats in scald+carnitine group were significantly lower than those in scald alone group (with t values of 34.40, 37.20, 19.95, and 7.88, respectively, P<0.01). At PIH 72, the protein expression levels of NLRP3, caspase-1, caspase-1/p20, GSDMD-N, and cleaved IL-1ß in liver tissue of rats in scald alone group were significantly higher than those in sham-injury group (with t values of 12.28, 26.92, 5.20, 10.02, and 24.78, respectively, P<0.01); compared with those in scald alone group, the protein expression levels of NLRP3, caspase-1, caspase-1/p20, GSDMD-N, and cleaved IL-1ß in liver tissue of rats in scald+carnitine group were significantly decreased (with t values of 10.99, 27.96, 12.69, 8.96, and 12.27, respectively, P<0.01). At PIH 72, the mRNA levels of GRP78 and CHOP in liver tissue of rats in scald alone group were significantly higher than those in sham-injury group (with t values of 21.00 and 16.52, respectively, P<0.01), and the mRNA levels of GRP78 and CHOP in liver tissue of rats in scald+carnitine group were significantly lower than those in scald alone group (with t values of 8.92 and 8.21, respectively, P<0.01); the protein expression levels of GRP78 and CHOP in liver tissue of rats in scald alone group were significantly higher than those in sham-injury group (with t values of 22.50 and 14.29, respectively, P<0.01), and the protein expression levels of GRP78 and CHOP in liver tissue of rats in scald+carnitine group were significantly lower than those in scald alone group (with t values of 14.29 and 5.33 respectively, P<0.01). After 24 h of culture, the cell survival rates of 0.1 µmol/L TM group, 0.2 µmol/L TM group, 0.4 µmol/L TM group, and 0.8 µmol/L TM group were significantly decreased than that in DMSO group (with t values of 4.90, 9.35, 18.64, and 25.09, respectively, P<0.01). Then 0.8 µmol/L was selected as the intervention concentration of TM. After 24 h of culture, compared with that in DMSO group, the protein expression levels of GRP78 and CHOP in cells in TM alone group were significantly increased (with t values of 10.48 and 17.67, respectively, P<0.01), and the protein expression levels of GRP78 and CHOP in TM+carnitine group were significantly lower than those in TM alone group (with t values of 8.08 and 13.23, respectively, P<0.05 or P<0.01). After 24 h of culture, compared with those in DMSO group, the protein expression levels of NLRP3 and GSDMD-N in cells in TM alone group were significantly increased (with t values of 13.44 and 27.51, respectively, P<0.01), but the protein expression levels of caspase-1, caspase-1/p20, and cleaved IL-1ß in cells were not significantly changed (P>0.05); compared with that in TM alone group, the protein expression levels of NLRP3 and GSDMD-N in cells in TM+carnitine group were significantly decreased (with t values of 20.49 and 21.95, respectively, P<0.01), but the protein expression levels of caspase-1, caspase-1/p20, and cleaved IL-1ß in cells were not significantly changed (P>0.05). Conclusions: In severely scald rats, exogenous L-carnitine may play a protective role against liver injury by inhibiting the pathways related to excessive endoplasmic reticulum stress-mediated pyroptosis.

[Influence of parental compliance on the treatment of hypertrophic scars in burn children].

Objective: To explore the influence of parental compliance on the treatment of hypertrophic scars in burn children. Methods: A retrospective cohort study method was used. From June 2014 to June 2019, 49 children with post-burn hypertrophic scars who met the inclusion criteria and visited the outpatient department of the Department of Burns of the First Affiliated Hospital of Anhui Medical University were included in this study. In the follow-up of 9 months, according to the registration form and the results of the compliance questionnaire for parents, the children were divided into good compliance group (34 cases, 21 males and 13 females, aged 2.0 (2.0, 3.5) years) and poor compliance group (15 cases, 6 males and 9 females, aged 3.0 (2.0, 4.0) years). At the first attendance and in the follow-up of 3, 6, and 9 months, the scar scores of children in good compliance group were evaluated by Vancouver Scar Scale (VSS). At the first attendance and in the follow-up of 9 months, the scar scores of children in poor compliance group were evaluated by VSS. At the first attendance and in the follow-up of 9 months, the scar pruritus scores of children in the 2 groups were evaluated by Verbal Rating Score (VRS). Data was statistically analyzed with chi-square test, Wilcoxon rank sum test, Mann-Whitney U test, independent sample t test, and paired sample t test. Results: At the first attendance, the color, vascular distribution, softness, and thickness scores, and total score in VSS scoring of scars of children in the two groups were similar (Z=0.834, 0.026, 0.837, 0.076, 1.074, P>0.05). In the follow-up of 9 months, the softness and thickness scores, and total score in VSS scoring of scars of children in good compliance group were significantly lower than those in poor compliance group (Z=5.518, 4.732, 5.042, P<0.01). Compared with those in the first attendance, the color, vascular distribution, softness, and thickness scores, and total score in VSS scoring of scars of children in good compliance group were significantly decreased in the follow-up of 9 months (Z=5.241, 5.273, 5.214, 5.245, 3.451, P<0.01); the color and vascular distribution scores, and total score in VSS scoring of scars of children in poor compliance group were significantly decreased in the follow-up of 9 months (Z=3.606, 3.542, 3.448, P<0.01). At the first attendance, the VRS score of scar pruritus of children in good compliance group was 6.00 (5.00, 6.25) points, which was similar to (5.47±1.69) points in poor compliance group (Z=0.607, P>0.05). In the follow-up of 9 months, the VRS score of scar pruritus of children in good compliance group was 1.00 (1.00, 1.25) points, which was significantly lower than (3.27±1.71) points in poor compliance group (Z=2.606, P<0.01). Compared with those in the first attendance, the VRS score of scar pruritus of children in good compliance group was significantly decreased in the follow-up of 9 months (Z=4.002, P<0.01), while there was no obvious change in poor compliance group in the follow-up of 9 months (t=3.550, P>0.05). Conclusions: Under the same treatment plan, good parental compliance has a positive effect on the treatment of hypertrophic scars in burn children decreasing the degree of scar hyperplasia and pruritus.

[Effects of early supplement of exogenous L-carnitine on renal function in severely scalded rats].

Objective: To explore the effects of early exogenous L-carnitine supplementation on renal function in severely scalded rats. Methods: According to the random number table, sixty-six adult female Sprague-Dawly rats were divided into healthy control group (n=6), scald alone group (n=30), and scald+ carnitine group (n=30). In the latter two groups, the rats were inflicted with full-thickness scald of 30% total body surface area on the back, and the lactated Ringer's solution was injected through the tail vein for resuscitation immediately after scald. At post injury hour (PIH) 1, rats in scald+ carnitine group were intraperitoneally injected with 100 mg/mL L-carnitine solution 400 mg/kg, while rats in scald alone group were intraperitoneally injected with the same volume of normal saline. Rats in these two groups were injected once every 24 hours thereafter. Six rats were taken from each of scald alone group and scald+ carnitine group to collect the renal tissue and abdominal aorta blood at PIH 6, 12, 24, 48, and 72, respectively. The serum content of total protein, albumin, urea nitrogen, creatinine, and cystatin C were determined by the automatic biochemical analyzer. Renal tissue was stained with hematoxylin-eosin to observe histopathological changes. Rats in healthy control group did not undergo any treatment, and their renal tissue and blood sample were extracted and analyzed in the same way as those of severely scalded rats. Data were statistically analyzed with one-way analysis of variance and Bonferroni method. Results: (1) The serum content of total protein and albumin of rats in scald alone group at each time point after injury was significantly lower than that in healthy control group (P<0.05). The serum content of total protein of rats in scald+ carnitine group was significantly higher than that in scald alone group at PIH 12 and 24 (P<0.05), and the serum content of albumin of rats in scald+ carnitine group was significantly higher than that in scald alone group at PIH 12 (P<0.05). The serum content of total protein and albumin of rats in scald alone group and scald+ carnitine group showed a trend of decrease followed by an increase, with the lowest value at PIH 24. (2) The serum content of urea nitrogen and creatinine of rats in scald alone group at each time point after injury was significantly higher than that of healthy control group (P<0.05). The serum content of urea nitrogen of rats in scald+ carnitine group was significantly lower than that in scald alone group at PIH 6, 48, and 72 (P<0.05). The serum content of creatinine of rats in scald+ carnitine group was significantly lower than that in scald alone group at PIH 12, 24, 48, and 72 (P<0.05). The serum content of urea nitrogen and creatinine of rats in scald alone group and scald+ carnitine group showed a trend of increase followed by a decrease, with the peak value at PIH 12. (3) The serum content of cystatin C of rats in scald alone group at PIH 6, 12, 24, 48, and 72 was (0.250±0.030), (0.330±0.070), (0.300±0.060), (0.240±0.060), and (0.190±0.030) mg/L, and the content at the first 4 time points were significantly higher than (0.170±0.020) mg/L of healthy control group (P<0.05). At PIH 24, the serum content of cystatin C of rats in scald+ carnitine group was (0.210±0.040) mg/L, which was significantly lower than that of scald alone group (P<0.05). The serum content of cystatin C of rats in scald alone group and scald+ carnitine group showed a trend of increase followed by a decrease, with the peak value at PIH 12. (4) The renal tissue of rats in healthy control group was almost normal, and the degree of renal tissue injury of rats in scald+ carnitine group was lighter than that in scald alone group at each time point after injury. At PIH 24, the renal tissue of rats in scald alone group showed extensive swelling of the renal tubular epithelial cells, vacuolar degeneration and necrosis, loss of brush borders, and nuclear shrinkage; more than 2/3 of the renal tubular cell nuclei disappeared, the tubular lumen was narrowed, necrotic exfoliated cells could be seen in the lumen, and edema and inflammatory cell infiltration could be seen in the renal interstitial. Compared with those of scald alone group, significantly reduced severity of edema and necrosis of renal tubular epithelial cells, as well as less inflammatory cell infiltration were observed in the renal tissue of rats in scald+ carnitine group. Conclusions: Early supplement of L-carnitine in severely scalded rats can reduce the damage of renal cells, accelerate the restoration of the content of total protein, albumin, urea nitrogen, creatinine, and cystatin C, thereby maintaining the stability of renal function metabolism level.

Circ_001680 aggravates the malignant process of gastric carcinoma by targeting MAP2.

OBJECTIVE: This study aims to explore the expression pattern and clinical significance of circ_001680 in gastric carcinoma (GC) process. PATIENTS AND METHODS: Circ_001680 levels in 40 pairs of GC and paracancerous ones were detected by quantitative real-time polymerase chain reaction (qRT-PCR). The relationship between circ_001680 and GC clinicopathological parameters was analyzed. AGS and SGC-7901 cells were used for constructing circ_001680 knockdown models by shRNA transfection. Proliferative and metastatic abilities in GC cells with circ_001680 knockdown were examined by cell counting kit-8 (CCK-8) and transwell assay, respectively. Dual-Luciferase reporter assay was conducted to clarify the interaction between circ_001680 and MAP2. Their co-regulation on GC process was detected through rescue experiments. RESULTS: Circ_001680 was highly expressed in GC tissues and cell lines. High level of circ_001680 predicted high incidences of lymphatic and distant metastasis, and poor prognosis in GC patients. Knockdown of circ_001680 suppressed proliferative and metastatic abilities in AGS and SGC-7901 cells. MAP2 was the target gene binding circ_001680, which was lowly expressed in GC. In addition, MAP2 was negatively correlated to circ_001680. Knockdown of MAP2 could abolish the suppressed proliferative and metastatic abilities in GC cells with circ_001680 knockdown. CONCLUSIONS: Circ_001680 is highly expressed in GC tissues and closely related to metastasis and prognosis in GC patients, which promotes the proliferative and metastatic abilities in GC cells by negatively interacting with MAP2.

[Analysis of chloroquine-resistant gene polymorphisms in Plasmodium falciparum imported into China in 2012 and 2018].

OBJECTIVE: To detect the chloroquine-resistant molecular marker polymorphisms in Plasmodium falciparum imported into China, investigate the mutation types of P. falciparum chloroquine resistant transporter (Pfcrt) gene at positions 72 to 76, and analyze the specificity of the P. falciparum specimens with different origins. METHODS: A total of 674 filter paper blood samples were collected from the National Malaria Diagnosis Reference Laboratory of China in 2012 and 2018. The amino acid po- sitions 72 to 76 of the Pfcrt gene on chromosome 7 were amplified using nested PCR assay and sequenced, and the sequencing results of the target gene fragment and the geographical region-specific prevalence of the mutations in the Pfcrt gene were analyzed. RESULTS: Among the 674 imported P. falciparum malaria cases in China in 2012 and 2018, 99.5% (644/674) were from Africa, which were predominantly from western and central Africa (80.4%, 518/644), and 4.5% (30/674) from Southeast Asia and Oceania (Papua New Guinea). A total of 4 site mutations (C72S, M74I, N75E and K76T) and 5 haplotypes (CVMNK, CVIET and SVMNT and two mixed types) were identified, with haplotypes CVMNK and CVIET present in parasites of both African and Southeast Asian origins, SVMNT detected in Southeast Asia (Myanmar) and Papua New Guinea isolates, the mixed type of haplo- types CVMNK/CVIET detected in P. falciparum of African and Southeast Asian origins, and the mixed type of haplotypes CVMNK/SVMNT detected only in the Myanmar isolate. Most P. falciparum parasites of the African origin carried the wild-type Pfcrt allele (77.7%, 478/615), and 68.0% (17/25) of the P. falciparum parasites of the Southeast Asian and Papua New Guinea or- igins harbored chloroquine resistant molecular markers (χ2 = 28.5, P < 0.05). The constituent ratio of the wild- and mutant-type Pfcrt allele varied in different geographical regions of Africa (P < 0.01), and the lowest prevalence of the wild-type Pfcrt allele was seen in western Africa. CONCLUSIONS: Among the 674 imported malaria cases in China in 2012 and 2018, the P. falciparum imported from Sotheast Asia habors a higher proportion of resistance to chloroquine and a higher molecular polymophism at ami- no acid positions 72 to 76 of the Pfcrt gene than the parasite of the African origin.

[Analysis of reasons for failure of Meek micro-skin grafting in children with severe burn and treatment measures].

Objective: To analyze the reasons for failure of Meek micro-skin grafting in children with severe burns and to observe the clinical effects of the treatment measures. Methods: Thirty children with severe burns hospitalized in the First Affiliated Hospital of Anhui Medical University (hereinafter referred to as the author's affiliation) from January 2012 to January 2018, conforming to the inclusion criteria were included to failed skin graft group. Children in failed skin graft group were performed with Meek micro-skin grafting operation and the operation failed, including 17 males and 13 females aged 1 to 12 year(s). Thirty children with severe burns hospitalized in the author's affiliation during the same period of time, conforming to the inclusion criteria, were included to successful skin graft group. Children in successful skin graft group were performed with Meek micro-skin grafting operation and the operation succeeded, including 16 males and 14 females aged 1 to 12 year(s). Main treatment measures and effects before operation, area and survival rate of Meek micro-skin graft, infected pathogens status, selection status of sensitive antibiotics, preoperative nutrition status, and wound infection status in plum rain season of children in the two groups, and nutritional status before and after strengthening nutritional support of postoperative surviving children in failed skin graft group were analyzed retrospectively. Data were processed with chi-square test and t test. Results: (1) The numbers of children in the two groups performed with main treatment measures of dilatation and anti-shock, tracheotomy intubation, ventilator-assisted respiration, and limb incision decompression after admission were close (χ(2)=0, 0.016, 0.025, 0.009, P>0.05). After taking the above-mentioned main treatment measures, effects of correcting shock, preventing asphyxia, correcting breathing difficulty, and improving peripheral circulation of limb were achieved. (2) The area of Meek micro-skin grafting of children in successful skin graft group was (20.6±2.5)% total body surface area (TBSA), close to (21.2±2.2)% TBSA in failed skin graft group (t=0.534, P>0.05). The survival rate of Meek micro-skin graft of children in successful skin graft group was (79±5)%, significantly higher than (26±3)% in failed skin graft group (t=2.956, P<0.01). (3) The microbial culture of wound secretion of 5 (16.67%) children in 30 patients in successful skin graft group was positive, with Pseudomonas aeruginosa of 2 children, and Escherichia coli, Staphylococcus aureus, and Aspergillus of one patient respectively. As children in successful skin graft group were with no symptom of systemic infection, no blood microbial culture was done. The microbial culture of wound secretion of 30 (100.00%) children in 30 patients in failed skin graft group was positive, and blood microbial culture of 8 (26.67%) children was positive. The main pathogen was Pseudomonas aeruginosa of 11 (36.67%) children in 8 pathogens caused infection with gram-negative bacteria of 22 (73.33%), gram-positive bacteria of 11 (36.67%) children, and fungi of 6 (20.00%) children. (4) Ten kinds of sensitive antibiotics such as cephalosporins, glycopeptides, carbapenems, and tetracyclines antibiotics were used in children in failed skin graft group, of which the use rate of imipenem of 9 (30.00%) was the highest. Only 4 kinds of sensitive antibiotics such as ceftazidime were used in 30 children in successful skin graft group. (5) The preoperative levels of albumin and prealbumin of children in successful skin graft group were (32±4) g/L and (133±41) mg/L respectively, significantly higher than (27±4) g/L and (93±35) mg/L in failed skin graft group (t=5.090, 4.064, P<0.01). The albumin and prealbumin levels of postoperative surviving children in failed skin graft group after nutritional support treatment were (35±4) g/L and (168±49) mg/L, significantly higher than (27±4) g/L and (94±38) mg/L before nutritional support treatment (t=6.911, 6.315, P<0.01). (6) Wound infection of 9 children in 30 children with wound infection in failed skin graft group happened in the plum rain season, and fungi infection of 3 children in 6 children with fungi infection happened in the plum rain season. Wound infection of 2 children in 5 children with wound infection in successful skin graft group happened in the plum rain season, and the only one children with fungi infection happened in the plum rain season. Conclusions: The main reasons for the failure of Meek micro-skin grafting in children with severe burns include infection, nutrition, and season factors, etc. Measures of strengthening wound dressing change, reasonable use of sensitive antibiotics to control infection, internal and external intestinal nutritional support, and reducing disturbance of the plum rain season by enhancing ventilation are effective and worthy of clinical promotion.

[Effect of early supplementation of exogenous carnitine on liver mitochondrial damage in severely scalded rats and its pathological mechanism].

Objective: To observe the effect of early supplementation of exogenous carnitine on liver mitochondrial damage in severely scalded rats and to explore its pathological mechanism. Methods: Seventy-two adult female Sprague-Dawley rats were divided into sham injury group, scald injury group, and scald injury+ carnitine group according to the random number table, with 24 rats in each group. Rats in sham injury group was sham injured on the back by immersing in 37 ℃ water bath for 12 s without fluid replacement. While rats in scald injury and scald injury+ carnitine groups were inflicted with 30% total body surface area (TBSA) full-thickness scald on the back by immersing in 98 ℃water bath for 12 s. Immediately after injury, rats in scald injury group and scald injury+ carnitine group were injected with Ringer's lactate solution with the dosage of 4 mL·kg(-1)·%TBSA(-1) via tail vein according to the Parkland formula, meanwhile rats in scald injury+ carnitine group were injected with L-carnitine solution with dosage of 300 mg·kg(-1)·d(-1) via tail vein from post injury hour (PIH) 1. At PIH 12, 24, 48 and 72, abdominal aorta blood and liver tissue were collected from 6 rats in each group. The serum levels of carnitine, ß-hydroxybutyric acid, and ornithine carbamoyltransferase (OCT) were determined with enzyme-linked immuno sorbent assay, and the serum levels of lactate dehydrogenase (LDH), alanine aminotransferase(ALT), and aspartate transaminase (AST) was determined by automatic biochemical analyzer, Pathological changes of rats liver tissue were detected with HE staining. Data were processed with analysis of variance of factorial design and Student-Newman-Keulstest or Tamhane test, Bonferroni correction. Results: (1) Compared with sham injury group, the serum level of carnitine of rats in scald injury group was significantly lower at each time point (P<0.05), and that of scald injury+ carnitine group was significantly lower at PIH 12, 24, and 48 (P<0.05). The serum level of carnitine of rats in scald injury+ carnitine group at PIH 72 [(28.2±3.0) µg/mL] was similar to that in sham injury group[(29.4±4.0) µg/mL, P>0.05]. The serum level of carnitine in scald injury+ carnitine group was significantly higher than that in scald injury group at each time point (P<0.05). (2) The serum levels of ß-hydroxybutyric acid of rats in scald injury group and scald injury+ carnitine group were significantly lower than those in sham injury group at each time point (P<0.05). The serum levels of ß-hydroxybutyric acid of rats in scald injury and scald injury+ carnitine groups both showed a trend of increase, and they peaked at PIH 72 [(1.77±0.30) , (2.93±0.44) mmol/L, respectively]. The serum levels of ß-hydroxybutyric acid in scald injury+ carnitine group were significantly higher than those of scald injury group at each time point (P<0.05). (3) The serum levels of OCT of rats in scald injury and scald injury+ carnitine groups were significantly higher than those of sham injury group at each time point (P<0.05). The serum levels of OCT of rats in scald injury group and scald injury+ carnitine groups both showed a trend of decrease, and they peaked at PIH 12 [(186.28±6.77), (163.38±9.34) ng/mL, respectively]. The serum levels of OCT of rats in scald injury+ carnitine group were significantly lower than those of scald injury group at each time point (P<0.05). (4) Compared with those of sham injury group, the serum levels of LDH of rats in scald injury group were significantly higher at each time point (P<0.05). Compared with those of sham injury group, those of scald injury+ carnitine group were significantly higher at PIH 12 and 24 (P<0.05), which peaked at PIH 12 [(2 226±274) U/L]. The serum levels of LDH of rats in scald injury+ carnitine group were close to those of sham injury group at PIH 48 and72 (P>0.05). The serum levels of LDH of rats in scald injury+ carnitine group were significantly lower than those of scald injury group at each time point (P<0.05). (5) The serum levels of ALT and AST of rats in scald injury group and scald injury+ carnitine group were significantly higher than those of sham injury group at each time point (P<0.05). In scald injury+ carnitine group, the serum levels of ALT of rats were significantly lower than those in scald injury group at PIH 48 and 72 (P<0.05), and the serum level of AST of rats was significantly lower than that in scald injury group at PIH 48 (P<0.05), and the serum levels of AST and ALT of rats were close to those in scald injury group at other time points (P>0.05). The serum levels of ALT and AST in scald injury+ carnitine group both showed a trend of decrease, and they peaked at PIH 12 [(260±25), (1 511±145) U/L, respectively]. (6) The liver tissue of rats in sham injury group was basically normal at each time point. The degree of liver injury of rats in scald injury+ carnitine group was lighter than that in scald injury group. The liver tissue of rats in scald injury group at PIH 72 showed obvious cytoplasm loose, liver tissue structure loss with diffuse fatty degeneration and large coagulative necrosis. Only partially scattered fatty degeneration was observed in the liver tissue of ras in scald injury+ carnitine group. Conclusions: By early supplementation of exogenous carnitine, serum levels of carnitine and ß-hydroxybutyric acid can be restored to normal levels faster, alleviate mitochondrial damage of hepatocytes, and maintain the metabolic stability of hepatocytes in early stage of severe scald.

[Influence of three-level collaboration network of pediatric burns treatment in Anhui province on treatment effects of burn children].

Objective: To explore the influence of three-level collaboration network of pediatric burns in Anhui province on treatment effects of burn children. Methods: The data of medical records of pediatric burn children transferred from Lu'an People's Hospital and Fuyang People's Hospital to the First Affiliated Hospital of Anhui Medical University from January 2014 to December 2015 and January 2016 to September 2017 (before and after establishing three-level collaboration network of pediatric burns treatment) were analyzed: percentage of transferred burn children to hospitalized burn children in corresponding period, gender, age, burn degree, treatment method, treatment result, occurrence and treatment result of shock, and operative and non-operative treatment time and cost. Rehabilitation result of burn children transferred back to local hospitals in 2016 and 2017. Data were processed with t test, chi-square test, Mann-Whitney U test, and Fisher's exact test. Results: (1) Percentage of burn children transferred from January 2014 to December 2015 was 34.3% (291/848) of the total number of hospitalized burn children in the same period of time, which was close to 30.4% (210/691) of burn children transferred from January 2016 to September 2017 (χ(2)=2.672, P>0.05). (2) Gender, age, burn degree, and treatment method of burn children transferred from the two periods of time were close (χ(2)=3.382, Z=-1.917, -1.911, χ(2)=3.133, P>0.05). (3) Cure rates of children with mild, moderate, and severe burns transferred from January 2016 to September 2017 were significantly higher than those of burn children transferred from January 2014 to December 2015 (χ(2)=11.777, 6.948, 4.310, P<0.05). Cure rates of children with extremely severe burns transferred from the two periods of time were close (χ(2)=1.181, P>0.05). (4) Children with mild and moderate burns transferred from the two periods of time were with no shock. The incidence of shock of children with severe burns transferred from January 2014 to December 2015 was 6.0% (4/67), and 3 children among them were cured. The incidence of shock of children with severe burns transferred from January 2016 to September 2017 was 3.9% (2/51), and both children were cured. The incidences and cures of shock of children with severe burns transferred from the two periods of time were close (χ(2)=0.006, P>0.05). Incidence of shock of children with extremely severe burns transferred from January 2014 to December 2015 was 57.1% (32/56), significantly higher than that of burn children transferred from January 2016 to September 2017 [34.5% (10/29), χ(2)=3.925, P<0.05]. Shock of 25 children with extremely severe burns transferred from January 2014 to December 2015 were cured, and shock of 9 children with extremely severe burns transferred from January 2016 to September 2017 were cured. The cures of shock of children with extremely severe burns transferred from the two periods of time were close ( χ(2)=0.139, P>0.05). (5) Time of operative treatment of children with moderate, severe, and extremely severe burns transferred from January 2014 to December 2015 was obviously longer than that of burn children transferred from January 2016 to September 2017 (t=2.335, 2.065, 2.310, P<0.05). Time of operative treatment of children with mild burns transferred from the two periods of time was close (Z=-0.417, P>0.05). Costs of operative treatment of children with moderate and severe burns transferred from January 2014 to December 2015 were significantly more than those of burn children transferred from January 2016 to September 2017 (Z=-3.324, t=2.167, P<0.05). Costs of operative treatment of children with mild and extremely severe burns transferred from the two periods of time were close (t=0.627, 0.808, P>0.05). (6)Time of non-operative treatment of children with mild, moderate, and severe burns transferred from January 2014 to December 2015 was obviously longer than that of burn children transferred from January 2016 to September 2017 (t=2.335, Z=-2.095, t=2.152, P<0.05). Time of non-operative treatment of children with extremely severe burns transferred from the two periods of time was close (t=0.450, P>0.05). Costs of non-operative treatment of children with moderate and severe burns transferred from January 2014 to December 2015 were obviously higher than those of burn children transferred from January 2016 to September 2017 (Z=-2.164, t=2.040, P<0.05). Costs of non-operative treatment of children with mild and extremely severe burns transferred from the two periods of time were close (t=0.146, 1.235, P>0.05). (7) Sixty-seven burn children transferred from January 2016 to September 2017 were transferred back to local hospitals for rehabilitation under the guidance of experts of the First Affiliated Hospital of Anhui Medical University, with 25 patients in 2016 and 42 patients in 2017. Effective rehabilitation rates of burn children transferred back to local hospitals for rehabilitation in 2016 and 2017 were both 100%. Conclusions: The three-level collaboration network of pediatric burns treatment in Anhui province can effectively increase cure rate of children with mild, moderate, and severe burns, reduce incidence of shock of children with extremely severe burns, shorten time of operative treatment of burn children with moderate, severe, and extremely severe burns, and time of non-operative treatment of children with mild, moderate, and severe burns, reduce treatment costs of children with moderate and severe burns, and improve rehabilitation effectiveness of children transferred from Lu'an People's Hospital and Fuyang People's Hospital to the the First Affiliated Hospital of Anhui Medical University.

[Effects of ω-3 polyunsaturated fatty acids on damage of intestinal mucosa of rats with severe burn in early stage and the mechanism].

Objective: To observe the effects of ω-3 polyunsaturated fatty acids (PUFA) on damage of intestinal mucosa of rats with severe burn in early stage and to explore the mechanism. Methods: One hundred and twenty SD rats were divided into sham injury group, pure burn group, and ω-3 PUFA group according to the random number table, with 40 rats in each group. Rats in sham injury group were sham injured, while rats in pure burn group and ω-3 PUFA group were inflicted with 30% total body surface area full-thickness scald (hereinafter referred to as burn) on the back. Rats in sham injury group and pure burn group were injected with normal saline solution (1 mL/kg) by tail vein, while rats in ω-3 PUFA group were injected with ω-3 PUFA solution (1 mL/kg) by the same way at 5 minutes post injury. At post injury hour (PIH) 3, 6, 12, 24, and 48, abdominal aorta blood and intestinal mucosa were collected from 8 rats in each group, respectively. Serum content of diamine oxidase (DAO) was detected by spectrophotography. Serum content of tumor necrosis factor alpha (TNF-α) and interleukin-6 (IL-6) was determined by enzyme-linked immunosorbent assay. Protein expression of NF-κB-p65 in intestinal mucosa was determined by Western blotting. Data were processed with analysis of variance of factorial design, one-way analysis of variance, chi-square test, LSD test, and Bonferroni correction. Results: (1) At all time points post injury, serum content of DAO of rats in pure burn group and ω-3 PUFA group was significantly higher than that in sham injury group (with P values below 0.01), and serum content of DAO of rats in ω-3 PUFA group was significantly lower than that in pure burn group (with P values below 0.01). (2) At all time points post injury, serum content of TNF-α and IL-6 of rats in pure burn group and ω-3 PUFA group was significantly higher than that in sham injury group (with P values below 0.01), and serum content of TNF-α and IL-6 of rats in ω-3 PUFA group was obviously lower than that in pure burn group (with P values below 0.01). (3) At all time points post injury, protein expressions of NF-κB-p65 in intestinal mucosa of rats in pure burn group and ω-3 PUFA group were significantly higher than those in sham injury group (with P values below 0.01). At PIH 3, 6, 12, 24, and 48, protein expressions of NF-κB-p65 in intestinal mucosa of rats in ω-3 PUFA group were 1.398±0.016, 1.999±0.948, 2.803±0.065, 1.739±0.602, and 1.484±0.645, obviously lower than 2.096±0.113, 3.402±0.189, 4.183±0.558, 3.618±0.408, and 2.614±0.775 in pure burn group (with P values below 0.01). Conclusions: The ω-3 PUFA may alleviate intestinal mucosa injury of rats with severe burn in early stage through reducing protein expression of NF-κB-p65 of intestinal mucosa, serum content of DAO, TNF-α, and IL-6, and inhibiting inflammatory response.

[Analysis on the performance evaluation of the Global Fund Malaria Programme in China from 2003 to 2013].

Objective: To analyze the performance of the 5 Global Fund Malaria Programmes in China from 2003 to 2013. Methods: All of the proposals, summaries, progress reports, survey reports, Monitoring& Evaluation reports, and performance rating reports of the 5 Global Fund Malaria Programmes in China and the epidemic data of program areas were collected for statistical analysis from 2012 to 2014. Symposiums were held with relevant experts from national and provincial Centers for Disease Control and Prevention, program managers and staffs from national and provincial Global Fund Malaria Programme offices. The completion of the relevant programme indicators (including the general grant information such as program areas, beneficiaries and funding; the implementation of malaria control measures; the performance of malaria control measures; the malaria incidence in the program areas; the prevalence of malaria parasites; and program management and performance evaluation) were analyzed, and the results: of the symposiums were summarized. Results The implementation period of the 5 Global Fund Malaria Programs were as follows: Round 1 from 2003 to 2008, Round 5 from 2006 to 2010, Round 6 from 2007 to 2012, Round 10 from 2012 to 2013, and National Strategy Application (NSA) from 2010 to 2012. Under the support of all the Global Fund Malaria Programs, a total of 11 936 726 fever cases received microscopic tests, 1 485 915 confirmed and suspected malaria cases were treated, 1 579 773 Long Lasting Insecticide-treated Nets were distributed, 3 414 633 regular nets were treated by insecticide, 40 298 284 primary and middle school students received health education on malaria control. Compared with the baseline value, the completion rates of each indicator increased after the implementation of the programs. The growth value ranged from 12.83% to 83.11%, among which the biggest growth was the value of the indicator'Percentage of households with at least one LLIN/ITN in target areas', and it increased from 9.2% (baseline value of 2006) to 92.31% (value of 2012). The malaria incidence in program areas has dropped significantly year by year, the annual reported malaria incidence in Yunnan and Hainan provinces decreased from 1 950/100 000, 3 850/100 000 in 2002 to 3.31/100 000, 0.15/100 000 in 2012, the P. falciparum malaria incidence in target counties in Hainan province decreased from 90.6/100 000 in 2002 to 0/100 000 in 2012. As from the implementation of NSA grant in 2010 to 2012, the annual reported malaria incidence in 92% of the 75 Type 1 counties was less than 1 per 10 000, 60.00% of Type 1 counties and 98.69% of the 687 Type 2 counties reported zero locally transmitted malaria cases. The Global Fund Secretariat had conducted a total of 37 performance evaluations, of which 9 have been rated as A1, 4 rated as A2, 19 rated as B1 and 5 rated as B1. Conclusions: The Global Fund Malaria Program in China has been closely integrated with the goal and task of National Malaria Control Program, reducing malaria burden in target areas, and pushing Chinese malaria control efforts to move from control to elimination.

Rituximab in combination with CHOP chemotherapy for the treatment of diffuse large B cell lymphoma in Chinese patients.

The objective of this study is to evaluate the long-term efficacy and safety of rituximab combined with cyclophosphamide, doxorubicin, vincristine, and prednisone (CHOP) in Chinese patients with newly diagnosed diffuse large B cell lymphoma (DLBCL). The study comprised a retrospective analysis of patients treated at a single center. Patients received four to six infusions of rituximab (375 mg/m(2) per dose) on day 1 of each cycle of CHOP chemotherapy. CHOP was initiated on day 3 of each cycle; cycles were repeated at 21-day intervals. A total of 82 patients with a median age of 45 years (range 18-76 years) was included. The overall response (OR) and complete response (CR) rates were 90.2 and 70.7%, respectively. The estimated 5-year progression-free survival (PFS) and overall survival (OS) rates were 56.4 +/- 8.3% and 74.1 +/- 7.4%, respectively. Patients with International Prognostic Index (IPI) scores < or = 2 had significantly higher OR, CR, PFS, and OS rates (p = 0.01, p = 0.02, p = 0.01, p < 0.001, respectively) compared with patients with IPI scores >2. The hematologic toxicity was mild. Five patients with a history of chronic hepatitis B experienced a reactivation of viral hepatitis. The rituximab-CHOP combination was effective and well tolerated in Chinese patients with newly diagnosed DLBCL.

Distribution of mRNA for the calmodulin-sensitive adenylate cyclase in rat brain: expression in areas associated with learning and memory.

The Drosophila learning mutant, rutabaga, is deficient in the calmodulin-sensitive adenylate cyclase, and studies of associative learning in Aplysia have implicated this enzyme in neuroplasticity. Therefore, the distribution of mRNA encoding the calmodulin-sensitive adenylate cyclase in rat brain was examined by in situ hybridization. mRNA for this enzyme is expressed in specific areas of brain that have been implicated in learning and memory, including the neocortex, the hippocampus, and the olfactory system. The presence of mRNA for this enzyme in the pyramidal and granule cells of the hippocampal formation provides evidence that it is found in neurons. These data are consistent with the proposal that the calmodulin-sensitive adenylate cyclase plays an important role in learning and memory.

A-type ATP binding consensus sequences are critical for the catalytic activity of the calmodulin-sensitive adenylyl cyclase from Bacillus anthracis.

Analysis of the predicted amino acid sequence of Bacillus anthracis adenylyl cyclase revealed sequences with homology to consensus sequences for A- and B-type ATP binding domains found in many ATP binding proteins. Based on the analysis of nucleotide binding proteins, a conserved basic amino acid residue in the A-type consensus sequence and a conserved acidic amino acid residue in the B-type consensus sequence have been implicated in the binding of ATP. The putative ATP binding sequences in the B. anthracis adenylyl cyclase possess analogous lysine residues at positions 346 and 353 within two A-type consensus sequences and a glutamate residue at position 436 within a B-type consensus sequence. The two A-type consensus sequences overlap each other and have the opposite orientation. To determine whether Lys-346, Lys-353, or Glu-436 of the B. anthracis adenylyl cyclase are crucial for enzyme activity, Lys-346 and Lys-353 were replaced with methionine and Glu-436 with glutamine by oligonucleotide-directed mutagenesis. Furthermore, Lys-346 was also replaced with arginine. The genes encoding the wild type and mutant adenylyl cyclases were placed under the control of the lac promoter for expression in Escherichia coli, and extracts were assayed for adenylyl cyclase activity. In all cases, a 90-kDa polypeptide corresponding to the catalytic subunit of the enzyme was detected in E. coli extracts by rabbit polyclonal antibodies raised against the purified B. anthracis adenylyl cyclase. The proteins with the Lys-346 to methionine or arginine mutations exhibited no adenylyl cyclase activity, indicating that Lys-346 in the A-type ATP binding consensus sequence plays a critical role for enzyme catalysis. Furthermore, the enzyme with the Lys-353 to methionine mutation was also inactive, suggesting that Lys-353 may also directly contribute to enzyme catalysis. In contrast, the protein with the Glu-436 to glutamine mutation retained 75% of enzyme activity, suggesting that Glu-436 in the B-type ATP binding consensus sequence may not be directly involved in enzyme catalysis. It is concluded that Lys-346 and Lys-353 in B. anthracis adenylyl cyclase may interact directly with ATP and contribute to the binding of the nucleotide to the enzyme.