Objective: Retro-laparoscopic partial nephrectomy is a challenge for ventro-renal tumors, especially hilar tumors.The tumors are partial or entirely out of operative field and there is blind space for operation.To solve this problem, a set of techniques including renal pedicle rotation is developed.Methods: A set of techniques including renal pedicle rotation, double-layer suture, early artery unclamping for retro-laparoscopic nephrectomy for ventro-renal tumors, especially hilar tumors were developed.Hilar tumors were located anteriorly to the angle of the renal and renal arteries and in contact with the anterior surface of the renal artery.The evaluation of the outcomes included the operation time, warm ischemia time, estimated haemorrhage, surgical margin, renal function after operation, and complications.This study reviewed the data of 28 patients with ventro-renal tumor, including 15 patients with complex hilar tumors.Results: The average tumor size was (3.8±0.5) cm.The average R.E.N.A.L score was 8 (ranging from 7 to 9).The average warm ischemia time was (23.8±4.7) min, and suture time was (26.2±6.5) min, and operation time was (124.1±12.1) min.The median hospital stay was 4 days and the average estimated haemorrhage was (139.1±54.0) mL.All the surgeries were operated under laparoscope and no surgery switched to open surgery.Seven patients developed Clavien Dindo Ⅰ-Ⅱ complications after operation.Three patients (10.7%) developed collective system damage and received repair of collective system in time and three patients (10.7%) developed slightly gross hematuria (Clavien-DindoⅠ).Two patients developed wound infection (Clavien-DindⅡ).Three patients (10.7%) got renal vein clamped because of capillary hemorrhage from basilar part of tumors.There were 14 patients received early unclamping of renal artery, and no obvious renal vessel damage was found.There were statistically significant differences between the serum creatinine levels (P<0.05) and the estimated glomerular filtration rates (P=0.02) before and after surgery.All the patients'' serum creatinine levels were in normal range and no patient needed regular dialysis.There was no positive surgical margin and the median follow-up was 7 months without local recurrence or distant metastasis.Conclusion: It is safe and effective to apply renal pedicle rotation technique in retro-laparoscopic partial nephrectomy for ventro-renal tumors.The short-term follow-up results are optimistic, but long follow-up is required.

Objective To explore the feasibility of electromagnetic navigation for hepatic ducts exploration by choledochoscope based on a three-dimensional (3D) printing model.Methods The retrospective descriptive study was conducted.The clinical data of 1 patient with obstructive jaundice combined with secondary biliary tract dilation who was admitted to the Beijing Tsinghua Changgung Hospital in April 2016 were collected.Digital imaging and standardized format data of enhanced computed tomography (CT) scan were collected,3D reconstruction of liver and bile duct were done under a 3D printing model,and then a hollow model of bile duct was achieved.Choledochoscope with internal electromagnetic probe was inserted into the printed hepatic ducts.Four points of anatomical markers in left and right hepatic ducts and common hepatic duct were chosen as fiducial markers for calibration and registration.Results After registration,the scope can be tracked in main hepatic segmental bile ducts.The locations of choledochoscope matched precisely the navigation results.Conclusion Electromagnetic navigation may bring accurate tracking effectiveness for choledochoscopic examination.

Objective:To investigate the efficacy and safety of cortical-sparing adrenalectomy (CSA) in the treatment of bilateral pheochromocytoma.Methods:The clinical data of 20 patients with bilateral pheochromocytoma treated in Xiangya Hospital of Central South University from January 2004 to December 2019 were analyzed retrospectively, including 10 males and 10 females. The average age of onset was 32.5 (8-51) years. 3 cases had a family history of pheochromocytoma. There were 14 and 6 patients with bilateral synchronous and metachronous onset, respectively. The mean value of vanilmandelic acid (VMA) in 20 cases was (106.4 ± 60.0) μ mol/24h. Preoperative enhanced CT showed a soft tissue mass with uneven enhancement in the adrenal region, with low-density necrosis, which suggested the diagnosis of Pheochromocytoma. All 20 cases underwent CSA under general anesthesia. In 14 cases of bilateral synchronous disease, 9 cases underwent simultaneous operation and 5 cases underwent staged operation; 6 patients with metachronous disease underwent bilateral tumor resection successively. Laparoscopic surgery was performed in 18 cases and open surgery in 2 cases. Through the abdominal or retroperitoneal approach, open the fat capsule around the upper pole of the kidney, free the medial edge of the upper pole of the kidney, expose the adrenal gland and tumor, completely remove the tumor and capsule, ensure that the adrenal tissue is 3-5 mm away from the cutting edge of the tumor, and the reserved cortical size is at least 1 / 3 of the ipsilateral adrenal gland. The central adrenal vein was preserved as much as possible to reduce the damage to the adrenal vascular bed. The operation related data, intraoperative monitoring records, postoperative complications and long-term follow-up results were recorded.Results:All the 20 cases were successfully completed without tumor rupture. The operation time of simultaneous operation and staged operation were (242.3 ± 61.0) min and (137.9 ± 60.3) min, respectively. The number of patients admitted to ICU after operation was 7 and 2, respectively ( P<0.05); The intraoperative bleeding volume was (528.6 ± 355.7) ml and (277.8 ± 264.7) ml, the number of blood transfusion cases were 5 and 2 cases, and the average hospital stay was (7.4 ± 2.0) d and (7.8 ± 3.3) d, respectively ( P>0.05). 20 cases took glucocorticoid orally (prednisone 5 mg, once every 12 hours) after operation. There was no obvious manifestation of adrenocortical dysfunction and Addison's crisis. The hormone was stopped gradually from 2 weeks to 1 month after operation. The average follow-up was 5.4 (1.0-16.0) years. There were 3 cases of recurrence and no metastasis. Gene detection was performed in 10 cases after operation, and 7 cases carried pheochromocytoma RET and VHL pathogenic gene mutations (RET in 2 cases and VHL in 5 cases). Conclusion:Although CSA has a certain risk of recurrence, it avoids hormone replacement and does not increase the risk of metastasis and death. It is recommended for the treatment of hereditary pheochromocytoma, especially bilateral pheochromocytoma.

Drug adulteration and contamination are serious threats to human health therefore,their accurate monitoring is very important.Allopurinol(Alp)and theophylline(Thp)are commonly used drugs for the treatment of gout and bronchitis,while their isomers hypoxanthine(Hyt)and theobromine(Thm)have no effect and affect the efficacy of the drug.In this work,the drug isomers of Alp/Hyt and Thp/Thm are simply mixed with α-,β-,y-cyclodextrin(CD)and metal ions and separated using trapped ion mobility spectrometry-mass spectrometry(TIMS-MS).TIMS-MS results showed that Alp/Hyt and Thp/Thm iso-mers could interact with CD and metal ions and form corresponding binary or ternary complexes to achieve their TIMS separation.Different metal ions and CDs showed different separation effect for the isomers,among which Alp and Hyt could be successfully distinguished from the complexes of[Alp/Hyt+y-CD+Cu-H]+with separation resolution(Rp-p)of 1.51;whereas Thp and Thm could be baseline separated by[Thp/Thm+y-CD+Ca-H]+with Rp-p of 1.96.Besides,chemical calculations revealed that the complexes were in the inclusion forms,and microscopic interactions were somewhat different,making their mobility separation.Moreover,relative and absolute quantification was investigated with an internal standard to determine the precise isomers content,and good linearity(R2＞0.99)was ob-tained.Finally,the method was applied for the adulteration detection where different drugs and urine were analyzed.In addition,due to the advantages of fast speed,simple operation,high sensitivity,and no chromatographic separation required,the proposed method provides an effective strategy for the drug adulteration detection of isomers.

OBJECTIVETo explore the molecular basis for three pedigrees affected with hypophosphatemia vitamin D resistant rickets (X-linked hypophosphatemia, XLH).

METHODSPeripheral blood samples from the three pedigrees were collected. Following DNA extraction, the 11 exons and flanking regions of the PHEX gene were subjected to PCR amplification and direct sequencing. Pathogenicity of identified mutations was evaluated through genotype-phenotype correlation.

RESULTSFor pedigrees 1 and 2, pathogenic mutations were respectively identified in exon 8 (c.871C>T, p.R291X) and exon 15 (c.1601C>T, p.P534L) of the PHEX gene. For pedigree 3, a novel mutation (c.1234delA, p.S412Vfs*12) was found in exon 11 of the PHEX gene, which caused shift the reading frame and premature termination of protein translation.

CONCLUSIONThe three mutations probably account for the XLH in the affected pedigrees. The discovery of novel mutations has enriched the spectrum of PHEX gene mutations.

Objective: To evaluate the pathogenicity of Acinetobacter venetum（Av），which is expected to be used as an environmental remediation agent，using Caenorhabditis elegans（C.elegans）. Methods: The C.elegans were cultured on the media loaded with E.coli OP50 and Av，respectively. The pathogenicity of Av was evaluated by observing the effects of Av on the growth，movement，digestive function，lifespan and reproduction of C.elegans，compared with that of another evaluation system according to NY 1109-2017 General Biosafety Standard for Microbial Fertilizers. Results: By C. elegans system，it was found that the body length，width，head thrash frequency，body bending frequency and average lifespan [（13.5±0.4）d vs.（13.7±0.4）d] of adult nematodes in the Av group were not significantly different from those in the OP50 group（all P>0.05）；while the average time of defecation cycle in the Av group shortened，the total number of progenies in the Av group increased by 18.7%（all P<0.05）. According to NY1109-2017 General Biosafety Standard for Microbial Fertilizers，it was found that the oral LD50 values for both male and female mice were more than 10 g/kgbw，which was practically non-toxic；the pathogenicity test of acute intraperitoneal injection showed that the animals did not have signs of poisoning，deaths or any abnormalities in gross anatomy；Av had no irritation to damaged skin and eyes of rabbits；the hemolysis test was negative；Av was sensitive to seven antibiotics and was medium to one antibiotic. Conclusion Av is not pathogenic. C. elegans can be used in early screening for the pathogenicity of environmental remediation agents.

OBJECTIVE: To explore the genetic basis for a fetus with cerebellar dysplasia and widened lateral ventricles. METHODS: The couple have elected induced abortion after careful counseling. Skin tissue sample from the abortus and peripheral venous blood samples from both parents were collected for the extraction of genomic DNA, which was then subjected to whole exome sequencing. Candidate variant was verified by Sanger sequencing. RESULTS: Prenatal ultrasonography showed increased nuchal translucency (0.4 cm) and widened lateral ventricles. Magnetic resonance imaging revealed infratentorial brain dysplasia. By DNA sequencing, the fetus was found to carry compound heterozygous variants c.1A>G and c.1564G>A of the RARS2 gene, which were inherited from its father and mother, respectively. Among these, c.1A>G was known to be pathogenic, but the pathogenicity of c.1564G>A was unreported previously. Based on the American College of Medical Genetics and Genomics guidelines, the c.1564G>A variant of RARS2 gene was predicted to be likely pathogenic(PM2+PM3+PP3+PP4). CONCLUSION The compound heterozygous variants c.1A>G and c.1564G>A of RARS2 gene contributed to the fetus suffering from pontocerebellar hypoplasia type 6, which expanded variant spectrum of RARS2 gene.
Female ; Fetus ; Genomics ; Humans ; Mutation ; Olivopontocerebellar Atrophies ; Pregnancy ; Whole Exome Sequencing

Objective:To explore the value of multi-parametric MRI for thyroid gland in differentiating benign and malignant thyroid nodules.Methods:From December 2018 to May 2020, 78 patients with 91 post-surgically pathologically confirmed thyroid nodules were enrolled in Cancer Hospital & Shenzhen Hospital, Chinese Academy of Medical Sciences and Peking Union Medical College. For each patient, the following MRI findings were obtained including the nodules′ location, size, shape, margin, signal intensity, cystic change, degree and pattern of contrast enhancement, involvement of surrounding structure, and ADC values. The time-intensity curve (TIC) were plotted and subtyped based on dynamic contrast enhancement MRI. The MRI findings between the benign and malignant thyroid nodules were compared using Mann-Whitney U test, χ 2 test or Fisher exact test. Multiple logistic regression analysis was used to select independent predictive variables and build a combined model, and the ROC curve was used to evaluate the diagnostic performance of each MRI finding and the combined model. Results:Between the benign and malignant thyroid nodules, the significant differences were found in size, shape, margin, presence of cystic changes, T 1WI signal intensity, ADC value, enhancement homogeneity, TIC subtypes and presence of thyroid capsule involvement ( P<0.05). Multivariate logistic analysis showed that ill-defined margin (OR=77.61), no presence of cystic changes (OR=36.11) and difference between TIC subtypes (OR=83.41) were independent predictive variables, and the area under the ROC curve (AUC) was 0.879, 0.788, and 0.751, respectively. The AUC, sensitivity and specificity of the combined model were 0.977, 0.986, and 0.904, respectively. Conclusions:Thyroid multi-parametric MRI derived findings can be used for the differential diagnosis of benign and malignant nodules. Combined with the independent risk factors with ill-defined margin, no presence of cystic changes, TIC of type plateau or washout, the diagnostic model has a higher diagnostic efficiency.

Objective: To establish the loop-mediated isothermal amplification (LAMP) method for detection of variola virus. Methods: One set of primers were designed for recognizing 5 distinct sequences on variola virus-specific gene HA. To optimize the reaction temperature and primers screening, and the sensitivity and specificity of this method for variola virus (VARV) detection were evaluated. Results: Rapid detection of variola virus by LAMP assay was completed within 60 min at 63 ℃. The sensitivity of LAMP with detection limits of 1 pg/μl was 10 times higher than that of PCR, that is, the LAMP sensitivity was 3.37×10⁵ copies/μl, and the PCR sensitivity was 3.37×10⁶ copies/μl. and the result of 2 kinds of other virus were negative, showing that it had a good specificity. Conclusions The method reported here demonstrates a potential and valuable means for detection of VARV. The LAMP assay is suitable for wide-area sample screening and on-site detection of variola virus in grassroots units, for on-site and primary quarantine, medical units for rapid diagnosis.

Objective    To explore the clinical applications of 3D-CT reconstruction combined with 3D printing in the analysis of anatomical types and variations of bilateral pulmonary arteries. Methods    From January 2019 to February 2022, the clinical data of 547 patients who underwent anatomical lung lesion resection in our hospital were retrospectively collected. They were divided into a 3D-CT reconstruction plus printing technology group (n=298, 87 males and 211 females aged 53.84±12.94 years), a 3D-CT reconstruction group (n=148, 55 males and 93 females aged 54.21±11.39 years), and a non-3D group (n=101, 28 males and 73 females aged 53.17±10.60 years). Results    In the 3D-CT reconstruction plus printing technology group, the operation time of patients (right: 125.61±20.99 min, left: 119.26±28.44 min) was shorter than that in the 3D-CT reconstruction group (right: 130.48±11.28 min, left: 125.51±10.59 min) and non-3D group (right: 134.45±10.20 min, left: 130.44±9.53 min), which was not associated with the site of surgery; intraoperative blood loss (right: 20.92±8.22 mL, left: 16.85±10.43 mL) was not statistically different compared with the 3D-CT reconstruction group (right: 21.13±8.97 mL, left: 19.09±7.01 mL), but was less than that of the non-3D group (right: 24.44±10.72 mL, left: 23.72±11.45 mL). Variation was found in the right pulmonary artery of 7 (3.91%) patients and in the left pulmonary artery of 21 (17.65%) patients. We first found four-branched lingual pulmonary artery in 2 patients. Conclusion    Preoperative CT image computer-assisted 3D reconstruction combined with 3D printing technology can help surgeons to formulate accurate surgical plans, shorten operation time and reduce intraoperative blood loss.