Accurate timing of myelination is crucial for the proper functioning of the central nervous system. Here, we identified a de novo heterozygous mutation in TMEM63A (c.1894G>A; p. Ala632Thr) in a 7-year-old boy exhibiting hypomyelination. A Ca²⁺ influx assay suggested that this is a loss-of-function mutation. To explore how TMEM63A deficiency causes hypomyelination, we generated Tmem63a knockout mice. Genetic deletion of TMEM63A resulted in hypomyelination at postnatal day 14 (P14) arising from impaired differentiation of oligodendrocyte precursor cells (OPCs). Notably, the myelin dysplasia was transient, returning to normal levels by P28. Primary cultures of Tmem63a^-/- OPCs presented delayed differentiation. Lentivirus-based expression of TMEM63A but not TMEM63A_A632T rescued the differentiation of Tmem63a^-/- OPCs in vitro and myelination in Tmem63a^-/- mice. These data thus support the conclusion that the mutation in TMEM63A is the pathogenesis of the hypomyelination in the patient. Our study further demonstrated that TMEM63A-mediated Ca²⁺ influx plays critical roles in the early development of myelin and oligodendrocyte differentiation.
Animals ; Cell Differentiation/physiology* ; Oligodendroglia/metabolism* ; Mice, Knockout ; Mice ; Male ; Myelin Sheath/metabolism* ; Humans ; Child ; Cells, Cultured ; Oligodendrocyte Precursor Cells/metabolism*

Objectives:To investigate the effect of the expression of low-density lipoprotein receptor associated protein (LDLR) on the vascular abnormalities in hepatocellular carcinoma (HCC) and its mechanisms.Methods:Based on the information of Oncomine Cancer GeneChip database, we analyzed the correlation between the expression level of LDLR and the expression level of carcinoembryonic antigen (CEA) and CD31 in hepatocellular carcinoma tissues. Lentiviral transfection of short hairpin RNA target genes was used to construct LDLR-knockdown MHCC-97H and HLE hepatocellular carcinoma cells. The differential genes and their expression level changes in LDLR-knockdown hepatocellular carcinoma cells were detected by transcriptome sequencing, real-time fluorescence quantitative polymerase chain reaction, and protein immunoblotting. The gene-related signaling pathways that involve LDLR were clarified by enrichment analysis. The effect of LDLR on CEA was assessed by the detection of CEA content in conditioned medium of hepatocellular carcinoma cells. Angiogenesis assay was used to detect the effect of LDLR on the angiogenic capacity of human umbilical vein endothelial cells, as well as the role of CEA in the regulation of angiogenesis by LDLR. Immunohistochemical staining was used to detect the expression levels of LDLR in 176 hepatocellular carcinoma tissues, and CEA and CD31 in 146 hepatocellular carcinoma tissues, and analyze the correlations between the expression levels of LDLR, CEA, and CD31 in the tissues, serum CEA, and alanine transaminase (ALT).Results:Oncomine database analysis showed that the expressions of LDLR and CEA in the tissues of hepatocellular carcinoma patients with portal vein metastasis were negatively correlated ( r=-0.64, P=0.001), whereas the expressions of CEA and CD31 in these tissues were positively correlated ( r=0.46, P=0.010). The transcriptome sequencing results showed that there were a total of 1 032 differentially expressed genes in the LDLR-knockdown group and the control group of MHCC-97H cells, of which 517 genes were up-regulated and 515 genes were down-regulated. The transcript expression level of CEACAM5 was significantly up-regulated in the cells of the LDLR-knockdown group. The Gene Ontology (GO) function enrichment analysis showed that the differential genes were most obviously enriched in the angiogenesis function. The Kyoto Encyclopedia of Genes and Genomes (KEGG) signaling pathway enrichment analysis showed that the relevant pathways involved mainly included the cellular adhesion patch, the extracellular matrix receptor interactions, and the interactions with the extracellular matrix receptors. The CEA content in the conditioned medium of the LDLR-knockdown group was 43.75±8.43, which was higher than that of the control group (1.15±0.14, P＜0.001). The results of angiogenesis experiments showed that at 5 h, the number of main junctions, the number of main segments, and the total area of the lattice formed by HUVEC cells cultured with the conditioned medium of MHCC-97H cells in the LDLR-knockdown group were 295.3±26.4, 552.5±63.8, and 2 239 781.0±13 8211.9 square pixels, which were higher than those of the control group (113.3±23.5, 194.8±36.5, and 660 621.0±280 328.3 square pixels, respectively, all P＜0.01).The number of vascular major junctions, the number of major segments, and the total area of the lattice formed by HUVEC cells cultured in conditioned medium with HLE cells in the LDLR-knockdown group were 245.3±42.4, 257.5±20.4, and 2 535 754.5±249 094.2 square pixels, respectively, which were all higher than those of the control group (113.3±23.5, 114.3±12.2, and 1 565 456.5±219 259.7 square pixels, respectively, all P＜0.01). In the conditioned medium for the control group of MHCC-97H cells,the number of main junctions, the number of main segments, and the total area of the lattice formed by the addition of CEA to cultured HUVEC cells were 178.9±12.0, 286.9±12.3, and 1 966 990.0±126 249.5 spixels, which were higher than those in the control group (119.7±22.1, 202.7±33.7, and 1 421 191.0±189 837.8 square pixels, respectively). The expression of LDLR in hepatocellular carcinoma tissues was not correlated with the expression of CEA, but was negatively correlated with the expression of CD31 ( r=-0.167, P=0.044), the level of serum CEA ( r=-0.061, P=0.032), and the level of serum ALT (r=-0.147, P=0.05). The expression of CEA in hepatocellular carcinoma tissues was positively correlated with the expression of CD31 ( r=0.192, P=0.020). The level of serum CEA was positively correlated with the level of serum ALT ( r=0.164, P=0.029). Conclusion:Knocking down LDLR can promote vascular abnormalities in HCC by releasing CEA.

Objectives:To investigate the effect of the expression of low-density lipoprotein receptor associated protein (LDLR) on the vascular abnormalities in hepatocellular carcinoma (HCC) and its mechanisms.Methods:Based on the information of Oncomine Cancer GeneChip database, we analyzed the correlation between the expression level of LDLR and the expression level of carcinoembryonic antigen (CEA) and CD31 in hepatocellular carcinoma tissues. Lentiviral transfection of short hairpin RNA target genes was used to construct LDLR-knockdown MHCC-97H and HLE hepatocellular carcinoma cells. The differential genes and their expression level changes in LDLR-knockdown hepatocellular carcinoma cells were detected by transcriptome sequencing, real-time fluorescence quantitative polymerase chain reaction, and protein immunoblotting. The gene-related signaling pathways that involve LDLR were clarified by enrichment analysis. The effect of LDLR on CEA was assessed by the detection of CEA content in conditioned medium of hepatocellular carcinoma cells. Angiogenesis assay was used to detect the effect of LDLR on the angiogenic capacity of human umbilical vein endothelial cells, as well as the role of CEA in the regulation of angiogenesis by LDLR. Immunohistochemical staining was used to detect the expression levels of LDLR in 176 hepatocellular carcinoma tissues, and CEA and CD31 in 146 hepatocellular carcinoma tissues, and analyze the correlations between the expression levels of LDLR, CEA, and CD31 in the tissues, serum CEA, and alanine transaminase (ALT).Results:Oncomine database analysis showed that the expressions of LDLR and CEA in the tissues of hepatocellular carcinoma patients with portal vein metastasis were negatively correlated ( r=-0.64, P=0.001), whereas the expressions of CEA and CD31 in these tissues were positively correlated ( r=0.46, P=0.010). The transcriptome sequencing results showed that there were a total of 1 032 differentially expressed genes in the LDLR-knockdown group and the control group of MHCC-97H cells, of which 517 genes were up-regulated and 515 genes were down-regulated. The transcript expression level of CEACAM5 was significantly up-regulated in the cells of the LDLR-knockdown group. The Gene Ontology (GO) function enrichment analysis showed that the differential genes were most obviously enriched in the angiogenesis function. The Kyoto Encyclopedia of Genes and Genomes (KEGG) signaling pathway enrichment analysis showed that the relevant pathways involved mainly included the cellular adhesion patch, the extracellular matrix receptor interactions, and the interactions with the extracellular matrix receptors. The CEA content in the conditioned medium of the LDLR-knockdown group was 43.75±8.43, which was higher than that of the control group (1.15±0.14, P＜0.001). The results of angiogenesis experiments showed that at 5 h, the number of main junctions, the number of main segments, and the total area of the lattice formed by HUVEC cells cultured with the conditioned medium of MHCC-97H cells in the LDLR-knockdown group were 295.3±26.4, 552.5±63.8, and 2 239 781.0±13 8211.9 square pixels, which were higher than those of the control group (113.3±23.5, 194.8±36.5, and 660 621.0±280 328.3 square pixels, respectively, all P＜0.01).The number of vascular major junctions, the number of major segments, and the total area of the lattice formed by HUVEC cells cultured in conditioned medium with HLE cells in the LDLR-knockdown group were 245.3±42.4, 257.5±20.4, and 2 535 754.5±249 094.2 square pixels, respectively, which were all higher than those of the control group (113.3±23.5, 114.3±12.2, and 1 565 456.5±219 259.7 square pixels, respectively, all P＜0.01). In the conditioned medium for the control group of MHCC-97H cells,the number of main junctions, the number of main segments, and the total area of the lattice formed by the addition of CEA to cultured HUVEC cells were 178.9±12.0, 286.9±12.3, and 1 966 990.0±126 249.5 spixels, which were higher than those in the control group (119.7±22.1, 202.7±33.7, and 1 421 191.0±189 837.8 square pixels, respectively). The expression of LDLR in hepatocellular carcinoma tissues was not correlated with the expression of CEA, but was negatively correlated with the expression of CD31 ( r=-0.167, P=0.044), the level of serum CEA ( r=-0.061, P=0.032), and the level of serum ALT (r=-0.147, P=0.05). The expression of CEA in hepatocellular carcinoma tissues was positively correlated with the expression of CD31 ( r=0.192, P=0.020). The level of serum CEA was positively correlated with the level of serum ALT ( r=0.164, P=0.029). Conclusion:Knocking down LDLR can promote vascular abnormalities in HCC by releasing CEA.

Objective:To analyze the risk factors of postoperative infection in patients with colon cancer, and construct a nomogram model.Methods:The clinical data of 220 patients with colon cancer in Anhui Cancer Hospital from May 2019 to June 2022 were retrospectively analyzed. Among them, 55 patients developed postoperative infection (infection group), and 165 patients did not develop postoperative infection (non-infection group). The receiver operating characteristic (ROC) curve was used to analyze the efficacy of each index in predicting postoperative infection in patients with colon cancer. Multivariate Logistic regression analysis was used to analyze the independent risk factors of postoperative infection in patients with colon cancer. R language 3.5.2 software was used to construct a nomogram model for predicting postoperative infection in patients with colon cancer, and it was verified and evaluated.Results:There were no significant differences in gender composition, body mass index, tumor stage, intraoperative blood transfusion, hypertension, smoking history, alcohol consumption history, tumor diameter and hemoglobin between the two groups ( P>0.05); the age, diabetes mellitus ratio, operation time and exhaust time in the infection group were significantly higher than those in the non-infection group: (49.60 ± 4.40) years old vs. (47.20 ± 4.12) years old, 63.64% (35/55) vs. 30.30% (50/165), (197.80 ± 12.55) min vs. (192.23 ± 12.05) min and (3.42 ± 1.18) d vs. (2.60 ± 0.80) d, the albumin was significantly lower than that in the non-infected group: (28.29 ± 3.02) g/L vs. (32.80 ± 3.21) g/L, and there were statistical differences ( P<0.01). ROC curve analysis result showed that the area under the curve of age, operation time, exhaust time and albumin for predicting postoperative infection in patients with colon cancer were 0.672, 0.610, 0.706 and 0.846, and the optimal cut-off values were 49 years old, 184 min, 3 d and 30 g/L, respectively. Multivariate Logistic regression analysis result showed that age (>49 years old), diabetes mellitus, operation time (>184 min), exhaust time (>3 d) and albumin (≤30 g/L) were independent risk factors of postoperative infection in patients with colon cancer ( OR = 2.131, 1.758, 1.449, 1.841 and 2.325; 95% CI 1.269 to 2.696, 1.354 to 3.059, 1.201 to 1.965, 1.018 to 2.365 and 1.582 to 3.051; P<0.01). A nomogram model was constructed with age, diabetes mellitus, operation time, exhaust time, and albumin as predictors for predicting postoperative infection in patients with colon cancer. The correction curve of the nomogram model for predicting postoperative infection in patients with colon cancer was close to the ideal curve (C-index = 0.764, 95% CI 0.657 to 0.834); decision curve analysis result showed that the nomogram model provided clinical net benefit when the risk threshold was > 0.07; and the clinical net benefit of the model was higher than that of age, diabetes mellitus, operation time, exhaust time and albumin. Conclusions:The age (>49 years old), diabetes mellitus, operation time (>184 min), exhaust time (>3 d) and albumin (≤30 g/L) are the independent risk factors of postoperative infection in patients with colon cancer, and the nomogram model based on the above variables could predict postoperative infection.

Objective:To investigate the clinical value of Octreotide plus Ulinastatin in the treatment of severe acute pancreatitis(SAP)in elderly patients.Methods:From May 2016 to February 2019, 124 elderly SAP patients admitted to the gastroenterology department of our hospital were enrolled and divided into the combination therapy group and the control group, with 62 patients in each group.The combination therapy group was treated with Octreotide and Ulinastatin, while the control group was treated with Octreotide alone.Serum leukocyte count, C-reactive protein(CRP), interleukin(IL)-6, tumor necrosis factor(TNF)-α and amylase levels were monitored before and 1, 3, 5, 7 and 14 days after treatment by automated biochemical analysis and enzyme-linked immunosorbent assays.The pain grade scale, APACHE-Ⅱ score and efficacy evaluation were analyzed for the two groups 7 days after treatment.The time to oral refeeding and length of hospitalization were compared between the two groups, and related complications during the treatment were recorded.Complications and the recurrence of pancreatitis within 1 year were followed up in both groups.Results:There was no statistically significant difference in serum white blood cell count, CRP, amylase, IL-6 and TNF-α levels between the two groups before treatment(all P>0.05). Serum white blood cell count, CRP and TNF-α levels had significant differences( t=3.735, 2.851 and -2.147, P=0.036, 0.029 and 0.043)and serum amylase and IL-6 levels had no significant difference( P>0.05)between the two groups 3 days after treatment.All the above parameters had significant differences between the two groups 7 days after treatment( t=3.624, 2.918, -2.166, 2.684 and -2.593, P=0.023, 0.011, <0.001, 0.015 and <0.001). Serum amylase, IL-6 and TNF-α levels had significant differences( t=-3.515, 4.627 and -3.189, all P<0.001)and serum white blood cell count and CRP had no significant difference(all P>0.05)between the two groups 14 days after treatment.There were significant differences in visual analogue scale(VAS)and APACHE-Ⅱ score between the two groups 7 days after treatment( t=-2.346 and -3.245, P=0.021 and 0.002). On the 7th day after treatment, the effectiveness rate was 79.0%(49/62)in the combination therapy group and 61.3%(38/62)in the control group, with a significant difference between the two groups( χ2=4.661, P=0.031). Compared with the control group, time to oral refeeding and hospitalization length were shorter in the combination therapy group than in the control group(6.72±1.87 d vs.7.65±1.69 d, 11.23±2.98 d vs.13.85±3.42 d, t=-2.868 and -4.565, both P<0.05). There were significant differences in the incidences of infectious pancreatic necrosis, gastrointestinal adverse reactions and organ failure between the combination therapy group and the control group(11.3% or 7/62 vs.25.8% or 16/62, 43.5% or 27/62 vs.21.0% or 13/62, 1.6% or 1/62 vs.11.3% or 7/62, χ2=4.324, 7.233 and 4.810, P=0.038, 0.007 and 0.028). There were significant differences in mean length of time without complications and recurrence between the combined group and the control group(10.25±3.26 months vs.8.72±3.73 months, 10.69±2.51 months vs.9.62±2.92 months, Log Rank χ2=7.463 and 4.589, P=0.006 and 0.032). Conclusions:Octreotide combined with Ulinastatin can effectively alleviate local symptoms, slow clinical progression, reduce the risk of complications, decrease the recurrence rate and promote early recovery in elderly SAP patients.

Objective:To analyze the variation and characteristics of gene mutation in patients with 6-pyruvoyltetrahydropterin synthase deficiency(PTPSD) in Henan province, and to provide the theoretical basis for early diagnosis, treatment, genetic consultation and prenatal diagnosis of PTPSD.Methods:One thousand nine hundred and six children with hyperphenylalaninemia (HPA) treated in Henan Neonatal Screening Center, the Third Affiliated Hospital of Zhengzhou University from January 1998 to December 2018 were included.Chemiluminescence was used for pheny-lalanine (Phe) detection in blood or dried blood spots.For patients with Phe concentration >120 μmol/L, urine pterin analysis was carried out, and the activity of dihydropteridine reductase (DHPR) was detected.Mutations of the PAH, GCH1, GFRP, PCBD1, PTPS and QDPR in 79 children with tetrahydrobiopterin deficiency(BH4D) were detected by using the high-throughput sequencing.All variations were verified by Sanger sequencing. Results:Among the 1 906 children, 79 cases were diagnosed as BH4D clinically, and they all were PTPSD.The incidence of PTPSD in HPA in Henan was 4.14%.One hundred and fifty-six out of 158 alleles in 79 children were detected, and the detection rate of gene mutation was 98.73%, 30 mutations were identified and most of the variants were located in exons 5(92/156 cases, 58.97%). Variants of c. 259C>T (61/156 cases, 39.10%), c.286G>A (17/156 cases, 10.90%), c.155A>G (13/156 cases, 8.33%) and c. 272A > G (10/156 cases, 6.41%) were more common.Six novel variations were detected, which included c. -77G>T, c.158A>G, c.262C>T, c.207G>A, c.316A>G and c. 332C>G; 38 genotypes had been identified, including 3 homozygous mutations and 33 compound heterozygous mutations.Conclusions:c. 259C>T is the hot-spots gene mutation in Chinese PTPSD patients in Henan province.The identification of 6 new mutations enriches the gene mutation profile.

Pulmonary hypertension (PAH) is a common pathological stats disease in clinic, it has a high morbidity and mortality.It was found that abnormal of endothelial growth factor, coagulation-related factors, 5-hydroxytryptamine, interleukin-6 and other inflammatory factors, hypoxia-inducible factors, transforming growth factor-β1 and connective tissue growth factor(CTGF) and so on were involved in the occurrence and development of PAH, but the specific mechanism was still unclear.In recent years, in monocrotaline(MCT) induced PAH rats model, they found that MCT rats had a significant higher concentration of mitochondrial coupling factor 6 (CF6) in both plasma and lung tissue than control group, which inhibited the activity of prostacyclin (PGI2), suggesting that CF6 may be involved in the occurrence and development of PAH.This paper will discuss the characteristics of CF6, the relationship between PGI2 and CF6, the relationship between PAH and CF6.

Objective To investigate the clinical value of serum procalcitonin (PCT) in diagnosis of urinary tract infection(UTI) in elderly patients.Methods 114 elderly patients with UTI in the department of infectious diseases of a hospital from January 2013 to December 2014 were analyzed retrospectively, clinical data of patients with abnormal and normal serum PCT were compared, PCT levels in patients with positive and negative blood cultures were compared, PCT receiver operating characteristic (ROC) curve for the diagnosis of bacteremia were drawn.Results Among 114 elderly patients with UTI, 46 were with abnormal PCT, 68 were with normal PCT.In abnormal PCT group, the proportions of patients with highest body temperature within 24 hours of admission, white blood cell count, neutrophil granulocyte percentage, C-reactive protein (CRP), blood urea nitrogen(BUN), creatinine(Cr), and urinary tract obstructive disease were all higher than those with normal PCT (all P<0.05).Among 42 patients with blood culture, PCT level in positive blood culture group(n=12) was higher than negative blood culture group(n=30)(1.93 [0.57-8.32] μg/L vs 0.36[0.15-1.01]μg/L, P=0.028).The area under the ROC curve (AUC) of the patients with bacteremia diagnosed by PCT was 0.72(95%CI:0.54-0.90),at the optimal value of 0.52 g/L, sensitivity, specificity, positive predictive value, and negative predictive value were 83.3%, 63.3%, 47.6%, and 90.5% respectively.Conclusion Serum PCT level can well reflect the severity of elderly patients with UTI, and is of great value in early diagnosis of bacteremia in elderly patients with UTI.

Objective To investigate serum interferon-inducible protein 16 (IFI16) level in the patients with preeclampsia (PE) and its correlation with PE pathogenesis.Methods Forty-five PE pregnant women in the Third Affiliated Hospital of Zhengzhou University from August 2015 to March 2016 were selected as the PE group and contemporaneous 30 healthy pregnant women undergoing the routine pregnant examination were selected as the control group.The biochemical indexes of serum urea,uric acid,creatinine,etc.were detected by using the automatic biochemical analyzer.The serum levels of IFI16 and ET-1 were measured by ELISA.Then the correlations between serum IFI16 level with these detected indicators were analyzed.The receiver operating characteristic (ROC) curve analysis was performed to evaluate the value of serum IFI16 for predicting PE disease.Results The serum IFI16 and ET-1 levels in the PE group were significantly higher than those in the control group (P<0.01).Furthermore the serum IFI16 level in severe PE was significantly higher than that in mild PE (P<0.01).Serum IFI16 level in PE was positively correlated with systolic blood pressure,diastolic blood pressure,24-h urine protein quantitation and serum ET-1 level,and negatively correlated with serum albumin.Serum IFI16 levels 14.47 ng/mL and 17.09 ng/mL as the critical values for predicting preeclampsia and discriminating between mild preeclampsia and severe preeclampsia has a higher sensitivity and specificity.Conclusion The high level of serum IFI16 in pregnant women has a certain correlation with PE pathogenesis and may be a novel biomarker for predicting PE occurrence.

Objective To investigate the antimicrobial resistance ,molecular phenotypes ,virulence gene profiles of Salmonella A gona (S .A gona) isolated from patients with acute diarrhea ,and to better understand its epidemic trend ,prevention and treatment .Methods Clinical data and stool samples of patients with acute diarrhea during April to October in 2013 and 2014 from the Second Hospital of Tianjin Medical University were collected .Enrichment culture ,biochemical identification and serotyping analysis were used to isolate and identify S .A gona strains .The isolated strains were further analyzed with antibiotics susceptibility test ,pulsed field gel electrophoresis (PFGE) ,multiple locus sequence typing (MLST ) , Quinolone resistance determining region (QRDR) .Plasmid-mediated quinolone resistance (PMQR) and β-lactamases genes (TEM ,SHV ,OXA ,and CTX-M) were amplified by polymerase chain reaction (PCR) and sequencing .The representative genes carried by Salmonella pathogenicity islands (SPI) 1 — 6 ,9 — 12 and virulence plasmids were amplified by PCR .And the clinical characteristics of S .Agona infection were analyzed .Results Among 119 non-repetitive (non-typhoidal salmonella ,NTS) isolates during the two years ,eight isolates (6 .7％ ) of S .A gona were identified . The resistance rate of S .A gona strains to streptomycin was 100 .0％ , those to ampicillin and gentamicin were 62 .5％ ,to levofloxacin ,ciprofloxacin and nalicixic acid were 25 .0％ ,to chloramphenicol ,amoxillin/clavulanic acid and piperacillin tazobactam were 12 .5％ .The strains were susceptible to other drugs .All 8 isolates had the identical ST13 genotype .PFGE showed 5 clones ,and 4 out of 5 isolates had the exact same patterns of PFGE and drug susceptibility .Two (fluoroquinolones ,FQ) resistant strains carried gyrA mutation leading to amino acid substitutions at position 87 in GyrA ,and no PMQR genes was detected ,while one of which was sensitive to ciprofloxacin by K-B method .All five ampicillin-resistant isolates were positive for TEM-1b gene and one isolate of them was resistant to β-lactam/β-lactamase inhibitor complex .The representative genes carried by SPI 1 — 6 , 9 ,11 ,12 (hilA ,sseL ,mgtC ,siiE ,sopB ,pagN ,bapA ,pagC and sspH2) were 100 .0％ positive ,while the genes carried by SPI10 (sef A ) virulence plasmids (spvB , prot6E) were negative . Two patients with FQ resistant strains infection were clinically diagnosed with bacillary dysentery ,and the remaining six cases with FQ susceptible strains infection were clinically diagnosed with acute gastroenteritis .Conclusions FQs-resistant and multi-drug resistant S .A gonaisolates have emerged in clinical settings .These isolates carry a variety of virulence genes .Resistance to FQ of S .Agonamay cause more severe illness .ST13 might be the dominant genotype of S . A gona in China ,and we should try to prevent the infection outbreak of S .A gona .