ObjectiveTo establish a analytical system for the survival motor neuron (SMN) subtle mutation,and evaluate its application in two families with spinal muscular atrophy (SMA).MethodsSMN genes in seven family members from two SMA families were analyzed at both transcript level and genomic level,by the use of the conventional PCR-RFLP,allele-specific PCR,multiplex ligation-dependent probe amplification (MLPA) and T subcloning and sequencing of SMNI gene.ResultsIn family A,the patient had a single SMN1 copy who was carrying nonsense mutation L228X,which was also found in his father.In family B,as the patient's sample was unavailable,the father was indeed a carrier with one normal SMN1 allele and the other SMN1 allele carrying a frameshift mutation 22_23insA.The remaining family members were SMA carriers with one SMN1 copy.ConclusionThis analytical system for SMN subtle mutation offers viable molecular basis for genetic counseling and prenatal diagnosis in SMA families.

Objective The aim of this study was to evaluate the left transthoracic approach in simultaneous antireflux surgery at the time of CDH repair in Infancy.Methods Between June 2008 and June 2012,18 patients underwent a left transthoracic approach in the treatment of type Ⅲ and Ⅳ hiatal hernia.,including 1 gastric volvulus.Clinical presentation in these patients included vomiting(n =12) 、pulmonary infections (n =9)、symptomatic anemia、failure to thrive (n =18).All the patients were evaluated before and after the surgery on clinical presentation,symptoms and functional assessment.Surgical techniques included extensive mediastinal esophageal dissection,Nissen fundoplication,resection of the hernial sac,crural closure.Results The average of operative time was 90 minutes.The mean blood loss was 5 ml.The average length of stay was 15 days.The hours stayed in PICU were 21.5 h.Mean follow-up was 22 months.There was no hospital mortality and hernia recurrence.Only one patient suffered gastroesophageal reflux disease after operation,and was controlled with antireflux medications.Conclusion Transthoracic is the optimal operative approach for treating the esophageal hiatal hernia,with relatively low postoperative morbidity and recurrence rates.Advantages of the transthoracic approach include the facilitation of Nissen fundoplication、esophageal lengthening procedures,and excellent exposure for the crural suturing.

Objective:To study the optimal pain control goal for preventing delirium in critical patients.Methods:A prospective cohort study were conducted. The patients admitted to general departments and transferred to the intensive care unit (ICU) due to critical illness in the First People's Hospital of Changde from January 2017 to November 2019 were enrolled. The General data of the patients were collected within 48 hours after admission. All patients admitted to the ICU were evaluated for pain level using the critical care pain observation tool (CPOT) every 8 hours by nurses, and confusion assessment method of ICU (CAM-ICU) was used to screen delirium patient every 8 hours by the leader of nursing team without knowing the pain level of the patients, until the subjects were transferred out of ICU. The receiver operating characteristic (ROC) curve was drawn, the area under ROC curve (AUC) and the optimal threshold were analyzed with delirium as the reference standard; according to the optimal threshold, multivariate Logistic regression analysis was used to evaluate the correlation between CPOT score and delirium.Results:During the study period, 575 patients were admitted to the participating departments and passed the preliminary screening according to the inclusion and exclusion criteria. During the study period, 34 patients were excluded due to incomplete data. Finally, a total of 541 patients were enrolled in the analysis, including 149 patients in delirium group and 392 patients in non-delirium group. There was no significant difference in gender, age, source of patients, education level, smoking history, drinking history, family mental history, acute physiology and chronic health evaluation Ⅱ (APACHEⅡ) score or other general information between the two groups. There were 10.1% (15/149) of patients in the delirium group used opioids, which was significantly higher than 4.3% (17/392) in the non-delirium group, and the difference was statistically significant ( P < 0.05). The CPOT score in the delirium group was significantly higher than that in the non-delirium group (4.24±1.78 vs. 2.75±1.95, P < 0.01). The patients were subdivided into young group (< 40 years old), middle-aged group (40-65 years old) and old group (> 65 years old) according to age. The analysis results were consistent with the overall analysis results. ROC curve analysis showed that the AUC of CPOT score predicting delirium was 0.719; when the best threshold value of CPOT score was 2.5, the sensitivity was 91.3%, the specificity was 49.0%, the positive predictive value was 40.5% and the negative predictive value was 93.7%. Multivariate Logistic regression analysis showed that the risk of delirium in ICU patients with CPOT score ≥ 3 was 10.043 times higher than that in patients with CPOT score < 3 [odds ratio ( OR) = 10.043, 95% confidence interval (95% CI) was 5.498-18.345, P < 0.001]. When the gender, age, APACHEⅡ score, smoking history, drinking history, opioids usage were adjusted, the risk of delirium in patients with CPOT score ≥ 3 was 10.719 times higher than that in patients with CPOT score < 3 ( OR = 10.719, 95% CI was 5.689-20.196, P < 0.001). Conclusion:The best pain control goal for preventing the occurrence of delirium in ICU patients is a CPOT score of 3 or less.

Objective To investigate methods for prenatal molecular diagnosis of fetuses at high risk for X-linked adrenoleukodystrophy(X-ALD).Methods The amniotic fluid was obtained and genomic DNA was isolated from amniotic fluid cells.Maternal contamination was evaluated by paternity test.PCRRFLP,sequencing and denaturing high performance liquid chromatography(DHPLC)were used to detect the ABCD1 gene of fetal genome.Results In the pedigree 1,the PCR product(799 bp)of the fetus 1 and her father(normal control)could be digested with BcnI. No P560L mutation,which was present in the index patient,was detected in the ABCD1 gene from the genomic DNA of the fetus 1 using direct sequencing.In the pedigree 2,the PCR product(232 bp)of the fetus 2 and her father could not be digested with MaeI and no Q177X mutation,which was present in the propositus,was detected in the ABCD1 gene from the genomic DNA of the fetus 2 using direct sequencing.In the pedigree 3,the PCR product(271 bp)was digested with AciI.the pattern of the fetus 3 and the propositus being the same,and the R617C mutation was found in the ABCD1 gene from the genomic DNA of the fetus 3 using direct sequencing.In the pedigree 4,the PCR product(269 bp)was analyzed with the DHPLC,and the pattern of elution peaks of the fetus 4 and her father was similar,but different from that of the propositus.No K276E mutation was detectable in the ABCD1 gene from the genomic DNA of the fetus 4 by using direct sequencing.Judging from the sex of the fetuses,fetuses 1 and 2 were normal homozygotes,fetus 3 was an ALD hemizygote,and fetus 4 was a normal hemizygote.Conclusion A new protocol for X-ALD prenatal molecular diagnosis is proposed,which would ensure the accuracy of prenatal diagnosis.

Purpose To investigate the histopathological features of cystic lung diseases ( CLD) , and to discuss the timing of clinical interventions. Methods HE and immunohistochemical staining were performed and reviewed in 125 cases of CLD. Results 125 ca-ses of CLD aged from birth to 11 years and 6 month, with an average age of 23. 0 months, median age 15 months, of which 60 cases were less than 1 year (48. 0%). 75 cases were male and 50 cases female, with male to female ratio of 1. 5 ∶ 1. Grossly, 50 cases showed single or multiple cysts with the size 0. 5 ~8. 0 cm in diameter, which did not communicate with bronchial cavity. 18 cases showed honeycomb cysts with the diameter of 0. 1~2. 0 cm. 26 cases were solid lesions without visible cysts. 21 cases were observed lung abscess with thick and rough wall and pus inside. 7 cases of emphysema showed microcysts with crepitation. 2 cases were identi-fied cystic and solid masses, with fish-fresh like cut surface. Histopathologically, 94 cases (75. 2%) were related to congenital bron-chopulmonary dysplasia in 125 cases of CLD, in which there were 59 patients (47. 2%) of congenial pulmonary airway malformation (CPAM), including 29 cases of type 1 (49. 2%), 18 cases of type 2 (30. 5%), and 12 cases of type 4 (20. 3%), there were 26 ca-ses (20. 8%) of pulmonary sequestration, including 15 cases of intralobar type (57. 7%) and 11 of extralobar cases (42. 3%), 5 ca-ses were complicated with CPAM type 2, 8 cases were bronchial cyst (6. 4%) and 1 case of enteric cyst (0. 8%). Acquired lesions were detected in 31 cases (24. 8%), including 21 cases of infected lung abscess, 1 case of fungal abscess. 7 cases of emphysema, and 3 cases of pleuralpulmonary blastoma (typeⅠ1 case and typeⅡ2 cases). Conclusion Pediatric CLD is characterized as com-plexed categories. The prognosis depends on correct pathological diagnosis, combined with imaging evaluation and appropriate timing of surgery.

Objective To investigate the clinical and pathological characteristics,diagnosis,differential diagnosis,treatment and prognosis of giant neurofibroma of penis in the child.Methods The clinical data including general data,imaging data,treatment methods,pathological characteristics of a case with giant neurofibroma of penis in a child were analyzed retrospectively and the relevant literature was reviewed.Results Gross appearance of the penile shaft neurofibroma was about 9 cm × 11 cm × 15 cm,with local ulceration.Computerized tomography scan revealed a giant mass in the penile shaft,about 9.0 cm × 10.0 cm × 13.4 cm.Partial excision of the penis was performed.Postoperative appearance of the residual penile shaft was about 2 cm long.The HE staining showed spindle cells with the red dye cytoplasm,spindle or elliptic nuclei and arranged in wavy partly.Positive immunostaining was presented with S-100 protein and Vimentin.The pathologic examination revealed a neurofibroma.There was no evidence of recurrence and the penis of the boy had normal sensation and erection by follow-up in 2 years.Conclusions Neurofibroma of penis in the child is extremely rare and the differential diagnosis of soft-tissue tumors of penis should be considered.The operative method should be individualized,the treatment goal is the complete resection;however,this goal must be weighed against detriment to functioning and the cosmetics of the involved organ.

Stem cells of various tissues including hematopoietic tissue in the body are derived from embryonic stem cells (ESC). There exists intricated gene regulation during ESC development and its differentiation into hematopoietic stem cells (HSC). In embryo hematopoiesis development, there are two kinds of hematopoietic types, primitive hematopoiesis and definitive hematopoiesis. The theory of the yolk sac of primitive hematopoiesis is well accepted, while the initial site of definitive hematopoiesis still exists controversy. In present opinion, there are at least two independent sites associated with definitive hematopoiesis, those are, yolk sac and para-aortic splanchnopleura (PAS)/aorta-gonad mesonephros (AGM). Study on the hematopoiesis and its regulation during embryonic ontogeny will benefit not only to the dicovery of the mechanism of some blood disorders, but as well to gene therapy and HSC engineering.

Objective:To evaluate the effect of surgery on 47 patients with moyamoya disease by retrospective analysis.Methods:A total of 47 patients with moyamoya disease were enrolled from August,2010 to According to the improved treatment in August,2013,all cases were divided into two groups:a pre-improved group and a post-improved group.According to different surgical methods,they were divided into two subgroups:an indirect revascularization subgroup and a combined revascularization subgroup.Results:The cerebral ischemia in 77.4％ of patients was relieved after the surgery.There was significant difference in outcomes of patients between the pre-improved group and the post-improved group (P＜0.05),while there was no significant difference between the pre-improved indirect revascularization subgroup and the pre-improved combined revascularization subgroup.There was also no significant difference between the post-improved indirect revascularization subgroup and the post-improved combined revascularization subgroups (P＞0.05).Conclusion:Surgical treatment can improve the outcomes of patients with moyamoya disease,but there is no significant difference in surgical effects between indirect and combined revascularization.

Objective To compare the clinical significance of peripheral blood neutrophil to lymphocyte ratio (NLR) and platelet to lymphocyte ratio (PLR) in predicting prognosis of patients with hepatocellular carcinoma (HCC).Methods The clinical data of 661 patients with HCC were retrospectively analyzed.Routine peripheral blood test results were used to calculate the NLR and PLR,and the receiver operating characteristic (ROC) curves were drawn.Using the thresholds of NLR and PLR,the patients were divided into the low NLR group and the high NLR group,and the low PLR group and the high PLR group.Overall survival (OS) and disease free survival (DFS) were evaluated by the Kaplan-Meier method.Independent prognostic predictors were determined by the Cox proportional hazard model.Results The NLR and PLR thresholds were 2.790 and 99,respectively.Analysis of the ROC curves showed higher NLR and PLR were significantly associated with poorer OS and DFS (all P ＜ 0.05).Multivariate analysis showed that NLR was an independent risk factor of OS and DFS (both P ＜ 0.05).The results remained unchanged when the NLR was further analyzed by applying different cut-off values of 2.810 and 3.In subgroup analysis,NLR remained an independent factor of Barcelona Clinical Liver Cancer staging system (BCLC) 0/A/B (P ＜ 0.05 for all measurements).Conclusion An elevated preoperative NLR could be a better prognostic predictor for HCC patients in comparison with PLR,especially for BCLC 0/A/B patients.

Objective To investigate the significance of albumin-bilirubin (ALBI) score in predicting prognosis of hepatocellular carcinoma (HCC).Methods The clinical data of 644 HCC patients were retrospectively analyzed from May 2010 to May 2013 in the First People's Hospital of Changde City and the Affiliated Tumor Hospital of Guangxi Medical University.Peripheral blood test results were used to calculate ALBI score,and ALBI score was categorized into the following 3 groups:grade 1,grade 2,grade 3.Overall survival(OS) and disease free survival(DFS) were evaluated by the Kaplan-Meier method.And independent prognostic predictors were determined by the Cox proportional hazard model.Results Multivariate analysis showed that ALBI grade 2 was an independent risk factor for OS(P＜0.05).The ALBI grade stratified patients into at least two distinct overall survival cohorts (P＜0.05),whereas the CP grade did not.The ALBI grade also classified patients with CP grade A patients into two distinct overall survival cohorts (P＜0.05).In the subgroup analysis,the ALBI grade seemed to be an independent factor in terms of Barcelona Clinical Liver Cancer staging system(BCLC) 0-C.Conclusion For patients with HCC who underwent R0 resection,the ALBI grade is a good predictor of OS in HCC patients,and the above conclusions is true in patients with BCLC stage 0-C.