Objective To evaluate the efficacy of proximal percutaneous pedicle screw fixation combined with distal open osteotomy for sagittal plane imbalance of adult spinal deformity.Methods From January 2011 to June 2015,23 patients with diagnosis of adult spinal deformity were treated with proximal percutaneous pedicle screw fixation combined with distal open osteotomy,there were 8 males and 15 females,aged from 52 to 67 years old (average,62.1 years old).The operation time,blood loss,drainage and perioperative complications were recorded;standing anteroposterior and lateral radiographs of the whole spine were taken and the following parameters were measured:sagittal vertical axis (SVA),lumbar lordosis(LL),pelvic tilt (PT),sacral slope (SS),pelvic incidence/lumbar lordosis mismatch (PI-LL),the above parameters were compared between preoperation and postoperation.Oswestry disability index (ODI) was used to evaluate the clinical efficacy.Results The mean operation time was 253.9±52.1 min,the mean blood loss and drainage was 1 258.5±272.2 ml and 725.1 ± 135.2 ml.No patient got infected,died or had deep vein thrombosis.All patients were followed up for an average of 21.2 months (range,13-52 m).The SVA was restored from 12.6±1.4 cm to 3.5±0.7 cm.In addition,LL,SS,PT,and PL-LL were improved from 13.5°±2.3°,13.9°±2.3°,29.7°±9.6°,29.5°±13.7° to 38.8°±9.6°,25.5°±5.8°,18.9°±8.2°,7.1°±3.6°.The ODI score decreased from 40.3％±12.5％ to 13.6％±2.57％ at the time of the last follow-up compared with preoperation.Conclusion Proximal percutaneous pedicle screw fixation combined with distal open osteotomy for sagittal plane imbalance of adult spinal deformity could restore the sagittal balance and improve the quality of life.

Objective To construct in the extracellular matrix metalloproteinase inducer (EMMPRIN)glycosylation single point mutation plasmid,and to explore its relationship with tumor cell proliferation.Methods PCR point mutantion technology was used to construct the mutantion plasimid of EMMPRIN glycosylation single point.After successful mutation, the function of mutantion plasmids were detected. Western blotting was used to detect the expression of EMMPRIN protein,immunofluorescence method was used to detemine the morphological changes of the cells,and MTT assay was performed to detect the relationship between mutantion pasmid and tumor cell proliferation.Results Confirmed by restriction enzyme digestion and sequencing,the 44th,the 152th,and the 186th Asn were successfully mutated to Gln in the sequence of EMMPRIN;EMMPRIN/GFP (N44Q), EMMPRIN/GFP(N152Q),and EMMPRIN/ GFP (N186Q)glycosylation single point mutation plasmids were constructed.Compared with wild-type,thel morphology of the cells was significantly changed,the core division of mutant-type cells was significantly reduced, the number of filopodia was reduced. The results of MTT assay showed that the survival rate of the cells in wild-type group were significantly increased compared with control group (P<0.05 );the survival rates of the cells in EMMPRIN (N44Q) group, EMMPRIN (N152Q) group and EMMPRIN(N186Q)were significantly decreased compared with wild-type group(P<0.05).Conclusion Mutant-type EMMPRIN can inhibit the proliferation of tumor cells;with the duration increasing, the inhibitory effect is weakened.There is a correlation between EMMPRIN glycosylation and proliferation of tumor cells.

Objective To assess the effects of axial spinous process-muscle-vascellum complex transplantation for posterior atlantoarial fusion.Methods Data of 27 cases with altantoarial disease who were treated by posterior atlantoarial fusion using axial spinous process-muscle-vascellum complex transplantation from June 2015 to June 2016 were retrospectively analyzed.There were 19 males and 8 females aged from 9 to 68 years old (mean,41.0±15.4 years old).Two cases were diagnosed with atlanto-axial instability.Fourteen cases were diagnosed with atlas fracture and eleven cases were diagnosed atlanto-axial fracture.All the 27 patients suffered from neck pain or limitations of cervical motion.All patients were assessed clinically by atlantoaxial reduction and bone graft fusion.The pre-operative and post-operative atlanto-dens interval (ADI),visual analogue scale (VAS),Japanese Orthopaedic Association scores (JOA),improvement rate of JOA score and axial symptoms were measured and statistically analyzed.Complications were recorded.Clinical outcome of latest follow-up was evaluated by X-ray and CT scan.Results The time of operation was 2.0-2.5 h and blood loss was 150-300 ml.All the patients were followed-up for 9 to 18 months (mean,11.5±2.1 months).The VAS of neck pain improved from 3.6±2.7 (range,2.0-5.0) pre-operatively to 1.4±0.2 (range,0-2.0) 12 months postoperatively (P=0.000).The JOA score improved from 11.7± 1.9 (range,10.0-15.0) pre-operatively to 15.3±0.6 (range,14.0-17.0)12 months post-operatively (P=0.000).The improvement rate of JOA score at the latest follow-up was 54.1％± 12.4％,including 23 cases (85.19％) excellent,and 4 cases (14.81％) good.The results of axial symptoms were no-symptom in 22 cases (81.48％) and mild symptoms in 5 cases (18.52％).Postoperative cervical spine X-ray and CT showed that the sagittal cervical spine alignment was restored.There was statistically significant difference between ADI of 4.3±1.1 mm (range,3.9-4.5 mm) pre-operatively to 2.5± 0.4 mm (range,2.1-2.6 mm) 12 months post-operatively,which was improved significantly (P=0.000).There were no complications found during the follow-up.Conclusion The application of axial spinous process-muscle-vascellum complex transplantation for posterior atlantoaxial fixation can preserve the dynamic function of muscles and reduce the postoperative pain,as well as avoid donor site morbidity.

Objective:To investigate the anatomical safety and feasibility ofposterior occipitocervical fixation with atlan-tooccipital-clivus screw.Methods:Data of 60 patients who treated in the spinal department of our hospital with upper cervical computed tomographic scans from February 2017 to November 2019 were retrospectively collected. Occipitocervical infection, injury, tumor and deformity were excluded. The Mimics software was used to reconstruct the occiput, atlas and measure the anatomical parameters, including the height and width of the anterior edge of the clivus, the height and width of the middle part of the clivus, the thinnest distance of the soft tissue in front of the clivus, the anteroposterior diameter, transverse diameter, the angle of inside tilting in coronary plane of the occipital condyle, the distance from the hypoglossal canal to the atlantooccipital articular surface, the anteroposterior diameter and transverse diameter of the superior joint of atlas, the height of the lateral mass, and the height and transverse diameter of the inferior articular process of the superior atlas joint. The three-dimensional digital modeling was performed and the screw diameter of 3.5mm was simulated. 3-Matic software were used to measure the screw placement parameters, including the inside tilting angle in coronary plane of screw, and the angle of upper tilting in sagittal plane and length of screw. The atlanto-occipital junction was exposed at the rear of 8 cadavers. According to the above parameters, the titanium alloy screws with a diameter of 3.5 mm were transferred from the inferior articular process and posterior arch of the atlas to the clivus through the atlantooccipital. Finally, the screw path was cut along the nail path with a pendulum saw, and the track of the screw was observed to confirm the safety and effectiveness of the screw.Results:The leading edge height and width of male clivus was 16.8±2.5 mm and 20.1±3.1 mm. The middle part of the clivus was 9.7±2.3 mm and 22.4±3.7 mm. The thinnest soft tissue in front of the clivus was 5.8±1.48 mm. The anteroposterior diameter of the occipital condyle was 19.1±1.9 mm, the transverse diameter was 12.6±2.0 mm, the inside tilting angle was 33.7°±4.5°, and the vertical distance from the lowest point of the neural tube to the articular surface of the occipital condyle was 9.6±1.1 mm. The height of the lateral mass of atlas was 12.9±2.4 mm, the anteroposterior diameter of the upper joint of atlas was 21.7±1.9 mm, and the transverse diameter was 11.7±1.4 mm. The width of the inferior facet was 14.9±1.4 mm and the height of the inferior facet was 5.7±0.85 mm. The distance from the screw entry point to the vertical line of the lateral mass migration midpoint was 2.5±0.6 mm; The distance from the screw entry point to the horizontal line of the midpoint was 2.3±0.7 mm.The inside titling angle of screw was 18.4°±1.6°, the upper tilting angle was 55.6°±3.1°, the length of the screw track was 53.0±2.8 mm, the adjustment range of upper tilting angle was 15.0±2.8 mm, the adjustment range of inside tilting angle was 10.4±2.4 mm. The anatomical parameters of females were slightly smaller than those of males, and the difference was statistically significant, but there was no significant difference between left and right parameters. The screws of 8 specimens could be inserted safely and effectively.Conclusion:Atlan-tooccipital-clivus screw can be implanted without damaging the nerve and vascular structure, and it can be used as a choice for occipitocervical fixation.

OBJECTIVE: To explore the genetic variation of a Chinese family affected with congenital insensitivity to pain with anhidrosis and albinism. METHODS: Whole exome sequencing (WES) was carried out to screen potential variants within genomic DNA extracted from the proband and his parents. Whole genome sequencing (WGS) was applied when variants were not found completely. Suspected variants were validated by Sanger sequencing. RESULTS: WES has identified a heterozygous c.1729G>C (p.G577R) variant of NTRK1 gene and two heterozygous variants of OCA2 gene, namely c.1363A>G (p.R455G) and c.1182+1G>A. WGS has identified two additional heterozygous variants c.(851-798C>T; 851-794C>G) in deep intronic regions of the NTRK1 gene. CONCLUSION The compound heterozygous variants of the NTRK1 gene probably underlay the congenital insensitivity to pain with anhidrosis. And the compound heterozygous variants of the OCA2 gene probably underlay the albinism in the proband. In the case where no variant is detected by WES in the coding region, WGS should be considered to screen potential variants in the whole genome.
Albinism ; Child ; DNA Mutational Analysis ; Hereditary Sensory and Autonomic Neuropathies/genetics* ; Heterozygote ; Humans ; Membrane Transport Proteins ; Mutation ; Pedigree

Objective:To explore the effect and mechanism of lncRNA SBF2-AS1 on glioma cell proliferation, apoptosis and radiosensitivity.Methods:Real-time quantitative polymerase chain reaction (RT-qPCR) was used to detect the mRNA expression levels of SBF2-AS1, miR-1287-5p and FSCN1 in normal human brain glial cell HEB and glioma cell lines, including LN18, SW1088, Hs683, and western blot method was used to detect the expression level of FSCN1 protein in cells. Glioma cells including LN18 and SW1088 were taken as the research object. After SBF2-AS1 small interfering RNA, miR-1287-5p mimics, FSCN1 small interfering RNA were transfected into LN18 cells, CCK-8 was used to detect cell proliferation, flow cytometry was used to detect cell apoptosis, clone formation experiment was used to detect cell radiosensitivity, Western blot was used to detect the protein expressions of cyclin D1, cleaved-caspase 3 and phosphorylated histone 2A variant phosphorylated histone (γ-H2AX). Dual luciferase reporter gene experiments verified the regulatory relationship between SBF2-AS1 and miR-1287-5p, as well as miR-1287-5p and FSCN1.Results:Compared with HEB cells, the expression level of SBF2-AS1 and the expression levels of FSCN1 mRNA and protein in glioma cell line LN18, SW1088 and Hs683 were significantly increased ( P<0.05), and the expression level of miR-1287-5p was significantly reduced ( P<0.05). After down-regulating SBF2-AS1, up-regulating miR-1287-5p or down-regulating FSCN1 expression, LN18 and SW1088 cells activity and cyclin D1 protein expression were significantly reduced ( P<0.05), the apoptosis rate and cleared-caspase-3 protein expression were significantly increased ( P<0.05), the survival score was significantly reduced ( P<0.05), and the expression ofγ-H2AX protein was significantly increased ( P<0.05). The results of dual luciferase reporter gene assay showed that the luciferase activity of LN18 and SW1088 cells co-transfected with miR-1287-5p mimics and SBF2-AS1-WT or FSCN1-WT was lower than that of co-transfected miR-NC and SBF2-AS1-WT or FSCN1-WT in LN18 and SW1088 cells ( P<0.001), while the luciferase activity of LN18 and SW1088 cells co-transfected with miR-1287-5p mimics and SBF2-AS1-MUT or FSCN1-MUT was not significantly different from that of miR-NC transfected with SBF2-AS1-MUT or FSCN1-MUT ( P>0.05). The expression level of miR-1287-5p in LN18 and SW1088 cells in si-SBF2-AS1 group was higher than that in si-NC group ( P<0.05), and the expression level of miR-1287-5p in LN18 and SW1088 cells in pcDNA-SBF2-AS1 group was higher than that in si-NC group ( P<0.05), but lower than that of pcDNA-NC group ( P<0.05). The protein expression level of FSCN1 in LN18 and SW1088 cells in the miR-1287-5p group was significantly lower than that in the miR-NC group ( P<0.05), and the protein expression level of FSCN1 in LN18 and SW1088 cells in the anti-miR-1287-5p group was significantly higher than that in the anti-miR-NC group ( P<0.05). When miR-1287-5p and SBF2-AS1 were down-regulated simultaneously or FSCN1 was up-regulated while SBF2-AS1 was down-regulated simultaneously, the proliferation and cyclin D1 protein expression of LN18 and SW1088cells were significantly increased ( P<0.001), the apoptosis rate and cleared-caspase-3 protein expression were significantly reduced ( P<0.001), the survival score was significantly increased ( P<0.001), and the expression of γ-H2AX protein was significantly reduced ( P<0.001). Conclusion:SBF2-AS1 highly expresses in glioma cells, down-regulation of SBF2-AS1 expression can inhibit the proliferation of glioma cells, promote apoptosis, and enhance cell radiosensitivity by regulating the miR-1287-5p/FSCN1 axis.

Objective:To explore the effect and mechanism of lncRNA SBF2-AS1 on glioma cell proliferation, apoptosis and radiosensitivity.Methods:Real-time quantitative polymerase chain reaction (RT-qPCR) was used to detect the mRNA expression levels of SBF2-AS1, miR-1287-5p and FSCN1 in normal human brain glial cell HEB and glioma cell lines, including LN18, SW1088, Hs683, and western blot method was used to detect the expression level of FSCN1 protein in cells. Glioma cells including LN18 and SW1088 were taken as the research object. After SBF2-AS1 small interfering RNA, miR-1287-5p mimics, FSCN1 small interfering RNA were transfected into LN18 cells, CCK-8 was used to detect cell proliferation, flow cytometry was used to detect cell apoptosis, clone formation experiment was used to detect cell radiosensitivity, Western blot was used to detect the protein expressions of cyclin D1, cleaved-caspase 3 and phosphorylated histone 2A variant phosphorylated histone (γ-H2AX). Dual luciferase reporter gene experiments verified the regulatory relationship between SBF2-AS1 and miR-1287-5p, as well as miR-1287-5p and FSCN1.Results:Compared with HEB cells, the expression level of SBF2-AS1 and the expression levels of FSCN1 mRNA and protein in glioma cell line LN18, SW1088 and Hs683 were significantly increased ( P<0.05), and the expression level of miR-1287-5p was significantly reduced ( P<0.05). After down-regulating SBF2-AS1, up-regulating miR-1287-5p or down-regulating FSCN1 expression, LN18 and SW1088 cells activity and cyclin D1 protein expression were significantly reduced ( P<0.05), the apoptosis rate and cleared-caspase-3 protein expression were significantly increased ( P<0.05), the survival score was significantly reduced ( P<0.05), and the expression ofγ-H2AX protein was significantly increased ( P<0.05). The results of dual luciferase reporter gene assay showed that the luciferase activity of LN18 and SW1088 cells co-transfected with miR-1287-5p mimics and SBF2-AS1-WT or FSCN1-WT was lower than that of co-transfected miR-NC and SBF2-AS1-WT or FSCN1-WT in LN18 and SW1088 cells ( P<0.001), while the luciferase activity of LN18 and SW1088 cells co-transfected with miR-1287-5p mimics and SBF2-AS1-MUT or FSCN1-MUT was not significantly different from that of miR-NC transfected with SBF2-AS1-MUT or FSCN1-MUT ( P>0.05). The expression level of miR-1287-5p in LN18 and SW1088 cells in si-SBF2-AS1 group was higher than that in si-NC group ( P<0.05), and the expression level of miR-1287-5p in LN18 and SW1088 cells in pcDNA-SBF2-AS1 group was higher than that in si-NC group ( P<0.05), but lower than that of pcDNA-NC group ( P<0.05). The protein expression level of FSCN1 in LN18 and SW1088 cells in the miR-1287-5p group was significantly lower than that in the miR-NC group ( P<0.05), and the protein expression level of FSCN1 in LN18 and SW1088 cells in the anti-miR-1287-5p group was significantly higher than that in the anti-miR-NC group ( P<0.05). When miR-1287-5p and SBF2-AS1 were down-regulated simultaneously or FSCN1 was up-regulated while SBF2-AS1 was down-regulated simultaneously, the proliferation and cyclin D1 protein expression of LN18 and SW1088cells were significantly increased ( P<0.001), the apoptosis rate and cleared-caspase-3 protein expression were significantly reduced ( P<0.001), the survival score was significantly increased ( P<0.001), and the expression of γ-H2AX protein was significantly reduced ( P<0.001). Conclusion:SBF2-AS1 highly expresses in glioma cells, down-regulation of SBF2-AS1 expression can inhibit the proliferation of glioma cells, promote apoptosis, and enhance cell radiosensitivity by regulating the miR-1287-5p/FSCN1 axis.

Objective:To investigate the clinical significance of a new classification system for atlas fractures based on pre- and post-treatment CT features, with a focus on diagnosis and treatment.Methods:A retrospective analysis was conducted on 75 cases of cervical vertebra fractures treated at the Sixth Hospital of Ningbo City between January 2015 and December 2020. The study included 44 males and 31 females, with an average age of 53.3±13.0 years (range: 27-81 years). The fractures were classified according to the Landells classification, resulting in 12 cases of type I, 13 cases of type II, 33 cases of type III, 9 cases that were difficult to classify due to fracture lines located at anatomical junctions, and 8 cases that could not be classified using the Landells classification due to diverse injury mechanisms. To establish a new preliminary classification for cervical vertebra fractures, the researchers considered whether the fracture line in the CT images involved the facet joint surface of the atlas, the impact on bilateral half-rings, and the displacement distance of the fracture ends. Five spinal surgeons were randomly selected to classify the CT images of the 75 patients using the new classification method. After one month, the imaging data of the 75 cases of cervical vertebra fractures were randomized and reclassified to assess the reliability and repeatability of the classification.Results:The new cervical vertebra fracture classification method comprised three types based on whether the fracture line involved the facet joint surface of the atlas: type A (no involvement of the facet joint surface of the atlas), type B (involvement of one side of the facet joint surface with intact contralateral half-ring), and type C (involvement of one side of the facet joint surface with fractured contralateral half-ring). Additionally, based on the maximum displacement distance between the fracture ends (>4 mm), six subtypes were identified: subtype 1 (≤4 mm displacement) and subtype 2 (>4 mm displacement). Consequently, the subtypes were classified as A1, A2, B1, B2, C1, and C2. According to the new classification method, the 75 patients included 17 cases of A1, 12 cases of A2, 7 cases of B1, 13 cases of B2, 12 cases of C1, and 14 cases of C2. The classification demonstrated excellent consistency, as assessed by the five doctors, with Kappa values of 0.85 and 0.91 for reliability and repeatability, respectively. At the final follow-up, all conservatively treated patients achieved bone healing, while four surgically treated patients experienced non-union of the fracture ends but exhibited good fusion between the atlas and axis. The remaining surgically treated patients achieved bony union without complications such as loosening or fracture of internal fixation.Conclusion:The new cervical vertebra fracture classification method, based on CT imaging features, comprehensively covers common clinical cases of cervical vertebra fractures and demonstrates excellent consistency. It provides valuable clinical guidance for the diagnosis and treatment of cervical vertebra fractures.

Objective:To investigate the safety of posterior atlantoaxial transarticular screw combined with atlas pedicle screw fixation in Chinese.Methods:CT data of upper cervical spine in 48 patients were collected from Ningbo No.6 Hospital, including 26 males and 22 females aged 26-58 years [(37.3±13.5)years]. All CT data was transformed into 3D dimensional model and inserted with virtual screws by Mimics 19.0. Firstly, the vertical plane P 1 and the horizontal plane P 2 of the atlas were built in those atlantoaxial models, secondly the atlantoaxial transarticular screw S 0 was inserted by Margel method, and it's insertion point was located at 3 mm lateral and 2 mm cephalad in the C 2 inferior articular process. Finally, four atlas pedicle screws were inserted at the midline of atlas lateral mass. The four screws are inserted as follows. S 1: the screw was tangent to the lateral side of the S 0 or the medial of the atlas pedicle. S 2: the screw was tangent to the lateral wall of the atlas pedicle. S 3: the screw was tangent to the upper wall of the atlas pedicle. S 4: the screw was tangent to the inferior wall of the atlas pedicle. The angles between S 1, S 2 and P 1 as the camber angle, and the angle between S 3, S 4 and P 2 as the gantry angle were measured. Then the safety range of camber angle and gantry angle were calculated, and the screw length of S 1, S 2, S 3 and S 4 was measured. In all models, the camber angle and gantry angle of the screws were adjusted an interval of 2°, the number of successful cases was calculated and the success rate of insertion was calculated. Results:All 3D models were inserted successfully. The minimum value, maximum value and safety range of the camber angle of atlas pedicle screws were (-6.7±5.2)°, (10.4± 4.3)°, (17.1±3.7)°, respectively; and of the gantry angle were (-0.5±3.5)°, (11.0±5.8)°, (11.5±4.9)°, respectively. The length of screw placement was S 1: (31.1±2.4)mm, S 2: (28.3±2.5)mm, S 3: (29.2±3.8)mm, S 4: (29.6±3.0)mm, respectively ( P<0.05). When the camber angle was from -1°to 5°, the success rate of screw placement was 87.5%; when inserting with 2° camber angle, the success rate was 100%; when the gantry angle was 5°, the success rate was 93.8%. Conclusion:The posterior atlantoaxial transarticular screw combined with atlas pedicle screw can achieve satisfy insertion length and success rate.

Objective:To compare the consistency of tomographic infrared spectrum analysis with conventional infrared spectrum analysis for the composition analysis of large-volume of urinary stones in vitro.Methods:Postoperative urinary stone specimens collected from 105 patients admitted to Beijing Tsinghua Changgung Hospital from January 2019 to June 2021 were analyzed, including 81 (77.14%) kidney stones, 16 (15.24%) ureteral stones, and 8 (7.62%) bladder stones. All stones measured ≥0.8 cm in maximum diameter on preoperative imaging. Eighty-four specimens, which were mainly stone fragments, were collected from percutaneous nephrolithotomy and ureteroscopic lithotripsy. These 84 specimens were analyzed and retested for stone composition using conventional infrared spectrum analysis by random multiple sampling. Other 21 renal stone specimens were obtained by laparoscopic lithotomy or standard percutaneous nephrolithotomy after November 1, 2020. These 21 specimens had a maximum diameter of ≥0.8 cm measured postoperatively. Based on intraoperative observation, stone specimens with typical layered structures were chosed. Then, all 21 samples were analyzed and retested by conventional infrared spectrum analysis and tomographic infrared spectrum analysis, respectively. When using tomographic infrared spectrum analysis, we need to take two maximum cross sections with a vertical spacing of these sections >2 mm, then perform multiple points sampling according to the morphological stratification of the first section. If the section's structure was homogeneous, we equidistantly took 2 to 3 samples from the center to the periphery. Otherwise, every layer needed to take a stone sample according to the stratification. Putting all the results of one section together, we obtained complete tomographic infrared spectrum analysis data. Take another coaxial cross-section of the same specimen for retesting. We recorded the characteristics of the three-dimensional distribution of stone composition in 21 stone specimens. Meanwhile, we compared the consistency of the results of conventional infrared spectrum analysis and tomographic infrared spectrum analysis for the same sample.Results:The consistency rate of the conventional infrared spectrum analysis was 56.19% (59/105), and that of tomographic infrared spectrum analysis was 80.95% (17/21). The difference in consistency between two methods was statistically significant ( χ2=4.447, P=0.035). Among 21 specimens, the consistency rate of conventional infrared spectrum analysis was 38.10% (8/21), which was significantly lower than that of tomographic infrared spectrum analysis ( χ2=7.814, P=0.005). Regarding the characteristics of the three-dimensional distribution of the components, the color and crystal morphology of five common types of stone components were different, and layered structure in the cross-section of the stones were observed. When the calculi were of the same composition, they were displayed in different morphology. We observed a trending change in the composition ratio between sublayers from the center to the edge in some compound-composition stones. Conclusions:For the composition analysis of larger-volume urinary stones, tomographic infrared spectrum analysis showed a higher consistency of retesting than conventional infrared spectrum analysis, and the three-dimensional distribution of stone composition had some characteristic features.