Using specimens spiked with adenovirus or porcine parvovirus, several DNA extraction methods were evaluated for their ability to remove polymerase chain reaction (PCR) inhibitors from stool samples. It was found that PCR inhibition could be partially overcome by extracting viral DNA with guanidine thiocyanate ( GuScN) or chelex resin methods, but PCR inhibition could be hardly overcome by extracting viral DNA with traditional proteinase-K phenol chloroform extraction method. For adenoviurs (dsDNA) in stool samples, GuScN, chelex resin, proteinase-K phenol chloroform methods respectively allowed 5 TCID50, 50TCID50, 50 TCID50 titer to be detected. For porcine parvovirus (ssD-NA) in stool samples, three protocols respectively allowed 10 TCID50, 50 TCD50, 100 TCID50 to be detected. . Their rapidity and low cost make GuScN and chelex resin extraction methods the suitable for routine diagnostic testing.

Objective To identify candidate regions of sporadic pheochromacytoma (PCC).Methods Totally 42 patients who were clinically diagnosed as sporadic PCC from 2011-9 to 2013-5 in PUMCH were enrolled .To extract whole genome DNA from their tumors as well as peripheral blood leukocytes and to exclude inherited cases by San-ger sequence mutation .Within 14 verified cases of sporadic PCC , we applyed single nucleotide polymorphism ( SNP) chip to detect whole genome DNA copy number variations ( CNV) and loss of heterozygosity ( LOH) to ini-tiallylocatethehotregions.FinallyapplyQ-PCRtoconfirmthehotregionsinleftcases.Results 38caseswere identified as sporadic PCC , and 4 as inherited cases .CNV were found in 14/14 tumors, of which deletions were more common.Missing regions occurred in 1p,3q,17p,22q,11p.On the other hand, of the 3 inherited cases, de-letion was also detected.The loss of parts of arm 1p is the most common, including chr1:74 957 006 ~86 132 879 , chr1:58 096 424~67 700 471 , and chr1:98 902 091~107 622 430 .The result of Q-PCR confirmed the above-mentioned three regions , and the three segments are final candidate regions .Conclusions Partially de-letion of 1p in most cases is the most striking phenomenon , we believe that the deletion of 1p may occur with PCC development , and there may be some tumor suppressor gene ( s) within these areas .

Objective To discuss the value of unilateral adrenalectomy(UA)for the treatment of refractory or recurrent Cushing disease(RRCD).Methods From January 2003 to July 2011,10 patients with RRCD underwent UA.The preoperative symptoms,preoperative treatments,intra-and post-operative complications and follow-up in patients were reviewed.All the 10 patients presented with hypercortisolemia symptoms.Transsphenoidal surgery had been performed in 9 patients as initial therapy.Of the 10 patients,8 received radiation therapy before adrenalectomy.All the 10 cases accepted a right UA.Results No perioperative complications occurred.The 10 patients were followed up for 1-8 years.Eight patients with radiation therapy before adrenalectomy got remission.Two bilateral adrenalectomies were eventually performed because the symptoms could not be controlled.Of the 8 patients who got remission,none of them developed Nelson syndrome and none need lifelong mineralocorticoid and glucocorticoid replacement therapy.Conclusions UA could be an option for RRCD.For patients with RRCD receiving radiation therapy,UA may effectively relief hypercortisolemia.Compared with bilateral adrenalectomies,UA could avert permanent adrenal insufficiency,which leads to a lifelong glucocorticoid and mineralocorticoid replacement therapies,and a life-threatening adrenal crisis.

Objective　To establish a rapid,effective,convenient method for detecting human adenoviruses and other viral pathogens using magnetic bead separation and PCR amplification.Methods　A biotinylated oligonucleotide primers were hybridized to adenovirus DNA in stool samples.Setreptavidin coated magnetic beads were then added to isolate the DNA-oligonucleotide hybrid.The procedure allows for the recovery of viral DNA suitable for amplification by polymerase chain reaction.Results　Ten samples collected from clinical stool were detected; six of them were positive.Results indicated that this nucleic acid separation technology is very effective in concentrating and purifying adenoviruses DNA while removing PCR inhibitors in stool samples.It also effectively increase the sensitivity of PCR amplification.Conclusion　This technique can rapidly,reliably detect adenoviruses in clinical samples,and it can be used to detect other viral pathogens.

Objective To discuss adrenalectomy effects on biochemical and metabolic impairment for patients with subclinical Cushing syndrome (SCS) due to an adrenal incidentaloma (AI) compared with conservative management.Methods Data from 87 patients with SCS due to AI in Peking Union Medical College Hospital between September 2011 and January 2016 were reviewed.Forty-eight patients underwent laparoscopic adrenalectomy (operative group), whereas 39 were managed conservatively (control group).No significant differences were found at baseline between the two groups concerning all the examined parameters, including age [(51.8 ± 10.2) years vs.(53.2 ± 12.1)years], male/female ratio (18/30 vs.20/19), size of adrenal mass [(3.01 ± 0.84) cm vs.(2.99 ± 1.01) cm], body mass index [(27.34 ±3.84) kg/m2 vs.(26.98 ± 3.99) kg/m2], adrenocorticotropic hormone (ACTH) [(2.01 ± 0.40) pmol/L vs.(1.96 ±0.42) pmol/L] and daily urinary free cortisol levels[(75.80 ± 39.71) mg vs.(71.06 ±31.35) mg], responsiveness to 1 mg of dexamethasone, arterial blood pressure, hypertensive patients (77.08％ vs.79.49％), glucometabolic control parameters, diabetic or impaired glucose tolerance patients (35.42％ vs.33.33％), lipid profile, dyslipidemia patients (70.83％ vs.71.79％).Results The duration of follow-up was 9-54 months in operative group, and 5-57 months in control group,respectively.In the operative group, laboratory corticosteroid parameters normalized in all patients but not in the control group.In the operative group, blood pressure of hypertensive patients improved or normalized (22 of 48).On the contrary, in the control group, cure or improvement was not achieved among the patients with hypertension, whereas worse condition was observed in 5 patients (P =0.004).No significant difference was found in glycemic control and blood lipid change between the two groups.However, a decrease in triglyceridaemia and HbA1c was found in operative group compared with the control group (P =0.011 and P =0.017, respectively).Substitutive corticosteroid treatment was administered in 3 patients due to postoperative adrenal insufficiency during hospital stay, and the duration of treatment was 9 weeks, 10 weeks and 12 weeks, respectively.Conclusion Laparoscopic adrenalectomy could be performed for patients with SCS due to AI.

Objective To study the initial experience and outcomes of laparoscopic enucleation with preoperative selective arterial embolization ( PSAE) for the treatment of renal angiomyolipomas. Methods A total of 43 patients with sporadic renal angiomyolipomas from July 2013 to November 2014 underwent laparoscopic nephron-sparing surgery ( NSS) .The patients were divided into either PSAE group ( n =19 ) or non-PSAE group ( n =24 ) .The data of patient demographics, success rate of surgery, perioperative complications rate, operating time ( OT ) , warm ischemia time ( WIT ) , estimated blood loss ( EBL ) , length of hospitalization, kidney estimated glomerular filtration rate (eGFR) and serum creatinine (Scr) level over 3 months, and ipsilateral recurrence were collected and compared between the two groups. Results Enucleation was successful in 100%patients (19/19) of the PSAE group and in 62.5%patients (15/24) of the non-PSAE group,with no significant difference ( Fisher' s test, P=0.105) The EBL was (46.4 ±20.6) ml in the PSAE group, which was significantly less than the non-PSAE group [(89.5 ±30.4) ml, t=-5.287, P=0.000].The OT and WIT were significantly shorter in the PSAE group than those in the non-PSAE group [(90.3 ± 21.1) min vs.(131.7 ±18.6) min, t=-6.831,P=0.000;(9.5 ±5.7) min vs.(24.2 ±4.8) min, t=-9.181, P=0.000]. The length of hospitalization was significantly longer in the PSAE group than that in the non-PSAE group [(7.7 ±1.1) d vs.(6.3 ± 1.3) d, t=3.748, P=0.000].No severe complications occurred in the PSAE group, whereas the non-PSAE group had 1 case of urinary leakage and 1 case of hematoma.The Scr level over 3 months after surgery was more ideal in the PSAE group [(70.1 ±13.7)μmol/L vs.(84.2 ±9.1) μmol/L, t=-4.045, P=0.000].No statistically significant difference was found in the eGFR between the PSAE group and the non-PSAE group [(72.6 ±12.8) ml· min-1· 1.73 m-2 vs.(68.0 ±10.7) ml· min-1· 1.73 m-2, t=1.284, P=0.206].No evidence of recurrence was found during follow-up period in both groups. Conclusions Laparoscopic enucleation with PSAE is a safe and effective minimally invasive procedure for the treatment of renal angiomyolipomas.As compared to traditional laparoscopic partial nephrectomy, it has advantages of less WIT, less EBL, and better protection of renal functions.It can be recommended in well-selected patients.

Objective To evaluate the diagnosis and treatment procedures for malignant pheochromocytoma. Methods 12 cases of malignant pheochromocytoma surgically treated were reviewed.This series comprised of 4 male and 8 female patients.All the tumors were confirmed on surgery and pathological studies. Results All the cases were operated.One died of hypotension after surgery.11 cases have been followed up for 2.5～13.0 years.4 died of recurrence and complication of hypertension.3 died of extensive metastases more then one year after operation.One died of pulmonary infection 4 years after operation and the others have been surviving. Conclusions Imaging procedures are important for the diagnosis of malignant pheochromocytoma.Recurrent tumors greater than 5 cm in diameter and uneven density in the tumor denote malignancy.Prompt radical extirpation of the tumor is the only means to achieve a long survival.

Objective To explore the viral pathogen spectrum characteristics of acute intestinal infection in Hanzhong from 2019 to 2022. Methods Fecal samples from patients with acute intestinal infection in the outpatient clinic of 3201 Hospital from January 2019 to December 2022 were collected. Common enteroviruses such as enterovirus 71 (EV-A71), coxsackievirus 16 (CV-A16), CV-A10, CV-A6, CV-A2, CV-A4, and CV-B3 were detected and analyzed by real-time fluorescence quantitative PCR. Results A total of 5 194 fecal samples were collected, and the positive rate of nucleic acid detection was 23.95%. In terms of the enteroviruses, the highest detection rate was 9.82% for EV-A71, followed by 4.58% for CV-A16 and 3.37% for CV-A6. The positive detection rate of common enteroviruses showed statistical difference among different age groups (P<0.05), with the highest detection rate of 41.49% in 0-4 years old group. There was no significant difference in the positive virus detection rate between different genders (P>0.05). EV-A71 infection showed no seasonal characteristics, whereas the detection of CV-A16 and CV-A6 infections was concentrated in summer and autumn. There were 106 cases of mixed infection, and the prevalence rate was 2.04%, with EV-A71 and CV-A6 mixed infections accounting for the majority of cases. Conclusion The main pathogens of acute intestinal infections in the Hanzhong area from 2019 to 2022 are EV-A71, CV-A16, and CV-A6. It is necessary to strengthen the monitoring of acute intestinal infections in children aged 4 years and below.

Objective: To investigate the value of enhanced recovery after surgery (ERAS) in D2 radical resection of distal gastric cancer under total laparoscopy and its effect on inflammatory factors. Methods: The clinical data of 50 patients with distal gastric cancer at Hanzhong Central Hospital from March 2016 to October 2017 were retrospectively analyzed. The patients were divided into the observation group (25 cases) and the control group (25 cases). The observation group received totally laparoscopic radical D2 gastrectomy for distal gastric cancer under the guideline of ERAS; the control group received totally laparoscopic radical D2 gastrectomy for distal gastric cancer under the guideline of the traditional method. The changes of perioperative related indexes, postoperative recovery and postoperative inflammatory indexes were compared between the two groups. Results: All 50 patients successfully completed the operation. The operation time was (287.2±52.3) min and (296.8±57.9) min, respectively of the observation group and the control group; the number of lymph node dissection was (34.0±11.0) and (34.6±14.3), respectively of the observation group and the control group. There were no significant differences in the above indexes between the two groups (all P > 0.05). The hospitalization time, the hospitalization cost, the postoperative time of getting out of bed of the observation group was (22.0±2.8) d, (61 044.4±4 692.7) yuan, (12.8±1.8) h. The hospitalization time, the hospitalization cost, the postoperative time of getting out of bed of the control group was (23.9±1.9) d, (63 875.81±4 392.6) yuan, (23.7±8.2) h. And there were statistically significant differences between the two groups (all P < 0.05). The pain score on the 1st day was (5.0±0.8) score and (6.4±1.0) score, and on the 4th day was (2.7±0.9) score and (3.9±0.9) score, respectively of the observation group and the control group, and there was a statistical difference (all P < 0.05). There was no statistical difference in the 7th day pain score between the two groups (P > 0.05). There were no statistically significant differences in the tumor necrosis factor α, interleukin-6 (IL-6), C-reactive protein and procalcitonin on the day before the operation of both groups (all P > 0.05); the levels of above four factors in the observation group were lower than those in the control group on the 1st, 4th and 7th day after the operation, and the differences were statistically significant (all P < 0.05). Conclusion The application of ERAS in totally laparoscopic radical resection of gastric cancer can improve the postoperative condition, significantly reduce the inflammatory reaction of patients, which has a certain value of improving the quality of life of patients.

【Objective】 To explore the role of ZFP36 in cardiomyocyte injury and autophagy induced by hypoxia/reoxygenation (H/R) so as to clarify its molecular regulatory mechanism. 【Methods】 H9C2 rat cardiomyocytes were infected with ZFP36 overexpressing lentivirus (OE-ZFP36) or its negative control lentivirus (OE-ZFP36 NC) to construct stable cell lines, respectively. Transfection of ATG4D overexpression plasmid (OE-ATG4D) improved the expression of ATG4D. Hypoxia/reoxygenation (H/R) induced myocardial cell injury. H9C2 cells were mainly divided into control group, H/R group, OE-ZFP36 NC+H/R group, OE-ZFP36+H/R group, OE-ATG4D NC+H/R group, OE-ATG4D+H/R group, OE-ZFP36+OE-ATG4D NC+H/R group, and OE-ZFP36+OE-ATG4D+H/R group. The protein expressions of ATG4D, Beclin1, LC3 and ZFP36 in H9C2 cells were detected by Western blotting. The mRNA levels of ZFP36 and ATG4D in H9C2 cells were detected by Real-time fluorescence quantitative PCR (qPCR). The viability of H9C2 cells was detected by CCK-8 assay. The levels of interleukin (IL-6) and tumor necrosis factor (TNF-α) in H9C2 cells were detected by enzyme-linked immunosorbent assay (ELISA). Reactive oxygen species (ROS) in H9C2 cells were detected by DCFH-DA method. SOD detection kit was used to detect the SOD level in H9C2 cells. The apoptosis of H9C2 cells was detected by flow cytometry. LC3 autophagosomes in H9C2 cells were detected by cellular immunofluorescence. Dual-luciferase reporter gene assay was used to detect the binding of ZFP36 and ATG4D mRNA in H9C2 cells. 【Results】 Compared with control group, H/R group showed decreased cell viability, increased IL-6 and TNF-α levels, increased ROS levels and decreased SOD levels, increased cell apoptosis. Up-regulated ATG4D and Beclin1 protein expression, increased LC3Ⅱ/LC3Ⅰ ratio, as well as upregulated ZFP36 expression were found in H/R group (all P<0.05). Compared with OE-ZFP36 NC+H/R group, elevated cell viability, decreased IL-6 and TNF-α levels, decreased ROS levels and increased SOD levels, reduced cell apoptosis (P<0.05), and downregulated ATG4D and Beclin1 protein expression, decreased LC3Ⅱ/LC3Ⅰ ratio were shown in OE-ZFP36+H/R group (all P<0.05). Compared with infection with OE-ZFP36 NC lentivirus, infection with OE-ZFP36 lentivirus decreased the luciferase activity of ATG4D 3′-UTR reporter gene, decreased the stability of ATG4D mRNA, and downregulated the H/R-induced ATG4D mRNA expression (all P<0.05). Compared with OE-ATG4D NC+H/R group, OE-ATG4D+H/R group had upregulated ATG4D mRNA and protein expression, decreased cell viability, increased IL-6 and TNF-α levels, increased ROS levels, decreased SOD levels and elevated cell apoptosis (all P<0.05). Compared with OE-ZFP36+OE-ATG4D NC+H/R group, OE-ZFP36+OE-ATG4D+H/R group had decreased cell viability, increased IL-6 and TNF-α levels, increased ROS levels, decreased SOD levels and elevated cell apoptosis (all P<0.05). 【Conclusion】 The expression of ZFP36 is upregulated in H/R-induced cardiomyocyte injury. The overexpression of ZFP36 inhibits H/R-induced cardiomyocyte injury and autophagy by regulating ATG4D, thus resisting cardiomyocyte H/R injury. It proves that ZFP36 is an important regulatory molecule against MI/RI.