Objective To evaluate the efficacy and complications of flexible ureteroscopy for the treatment of upper urinary tract calculi in children.Methods The clinical data of upper urinary tract calculi in 20 children including 16 male and 4 female,treated by using flexible ureteroscopy,were collected from January 2014 to May 2015 in the First Affiliated Hospital of Zhengzhou University.The mean age was 6.4 years (ranging from 2.7-15.0 years old).Among them,6 cases had upper ureteral calculi and 14 cases had renal calculi.All the calculi was found unilaterally with on the left side in 12 cases and on the right side in 8 cases.Ipsilateral mild to moderate hydronephrosis were found in all of the cases.The mean stone size was 1.2 cm (ranging from 0.5-1.8 cm).All the cases were treated through retrograde flexible ureteroscopy holmium laser lithotripsy after the invalid conservative through conservative treatment.Double J tube was left routinely in the sick side 1 week before operation to expand the internal diameter of the ureter.A double J stent was left in place for 1 month after operation as a routine.Results The flexible ureteral access sheath failed to insert into the upper ureter in 2 cases because of the narrow ureter,then the flexible ureteroscopy was inserted into ureter through wire directly.The flexible ureteral access sheath was successfully inserted into the ureter for the others and flexible ureteroscopy were inserted through flexible sheath.The mean operative time was 38 min (ranging from 20-60 min).The patients were discharged from hospital after a mean of 4.5 days (ranging from 3-7 days).The rate of stone-free in one-stage was more than 90％.There were residual small stones in the lower calices of kidney in 2 cases.The perfusion liquid volume during the operation was 500 mL(ranging from 200-1 000 mL).There was no major perioperative complication,while transient macroscopic hematuria and fever were common complications after operation.The stone search was performed successfully in the whole 14 cases.NO residual stone could be found through ultrasound and kidney ureter bladder plain and Double-J stent was removed 1 month after operation.Conclusions Flexible ureteroscopy produces high stone-free rate and clinical safety for upper urinary calculi in children.However,the complications of flexible ureteroscopy lithotripsy cannot be ignored and sufficient preoperative preparation and careful perioperative safety control should be required.

OBJECTIVE:To establish a method for simultaneous determination of 4 psoralen compounds in Buwu tincture. METHODS:HPLC was performed on the column of Dikma Diamonsil C18 with mobile phase of acetonitrile-0.2% Acetic acid by gradient elution at flow rate of 1 ml/min,detection wavelength was 246 nm,column temperature was 30 ℃,and the injection vol-ume was 15 μl. RESULTS:The linear range was 13.00-208 μg/ml for angelicin,26.00-416 μg/ml for bavachin,24.50-392 μg/ml for psoralidin and 37.88-606 μg/ml for isobavachalcone,respectively(r≥0.999 6);RSDs of precision,stability and reproducibility tests were less than 2.00%;recoveries were 95.22%-97.23%(RSD=0.87%,n=6),100.24%-104.64%(RSD=1.62%,n=6), 102.28%-104.39%(RSD=1.47%,n=6)and 97.68%-100.17%(RSD=0.97%,n=6),respectively. CONCLUSIONS:The method is simple and reproducible,and can be used for the quality control of Buwu tincture.

Objective To analyze the complexity of molecular structure in porcine T cell receptor gene and its similarity compared to humans.Method Based on the gene of porcine T cell receptor alpha chain ( TCRα) from the Gen-Bank database, 93 swine T cell receptor alpha chain genes ( STA) were cloned by reverse transcription-polymerase chain reaction (RT-PCR) from porcine peripheral blood lymphocytes, lymph nodes and spleen.Result Sequence analysis showed that STA genes all contain a domain of variable signal peptide and V, hypervariable J and conservative C.Howev-er, nucleotide sequence of STA was not completely identical with only 68.4%to 98.7%homology among genes, and had extremely sophisticated polymorphism and diversity.This was accord with the genetic structure of TCRαchain.Molecular structure, genetic evolution and classification of these genes were carried out according to the homology of TCRαgene, which all have several sites and zones of mutation on the domain of signal peptide, FR1 and CDR1, FR2 and CDR2, FR3 and CDR3.Analysis of similarity and classification of TCRαV domain(STAV)and J domain (STAJ) of Hezuo minipig u-sing IMGT/V-QUEST tools compared with those of humans found the genetic evolution relationship that was closer, and each of TRAV and TRAJ also found to have a corresponding fragment of humans, ever in 92% of similarity of TRAV be-tween swine and humans.Conclusion Our results indicate that inbred Hezuo minipig possesses genetic diversity against complicated environment of microbes in healthy status, and Hezuo minipig is suitable as an animal model for research on human immunology and diseases.

Background and purpose:Δ133p53 can promote tumor cell growth, but the exact mechanism is not clear. This study was aimed to observe the expression and signiifcant of the p53 isoformsΔ133p53 and p53 gene downstream molecules MDM2 and cyclin G1 genes by 5-FU-MKN45 gastric cancer cell line model. Methods:Af-ter using different concentrations of 5-FU (50μg/mL, 100μg/mL) to human gastric cancer cell line MKN45, inhibition rate should be detected by MTT assay, the changes ofΔ133p53 mRNA, MDM2 mRNA and cyclin G1 mRNA express-ing were detected by reverse transcription-polymerase chain reaction (RT-PCR). Differences between these groups were analyzed by ANOVA, comparisons within groups were analyzed by t-test, bivariate correlation was analyzed by Pearson linear correlation. Results:MTT results showed that with the increased concentration of 5-FU and the extension of time, the cell inhibition rates increased gradually. The inhibition rates of 50μg/mL 5-FU were 41.10%, 54.79%and 68.48%, for culturing 24, 48 and 72 hours. There were statistically signiifcant differences between the groups(F=45.52, P=0.00). The inhibition rates of 100μg/mL 5-FU were 69.53%, 78.21%and 86.92%, for culturing 24, 48 and 72 hours. There were statistically signiifcant differences between the groups(F=85.58,P=0.00). The inhibition rates of 50 and 100μg/mL were 41.10%and 69.53%, for culturing 24 hours. There were statistically signiifcant differences between the groups(F=51.29, P=0.00). The inhibition rates of 50 and 100μg/mL were 54.79%and 78.21%, for culturing 48 hours. There were statistically signiifcant differences between the groups(F=51.29, P=0.00). The inhibition rates of 50 and 100μg/mL were 68.48%and 86.82%, for culturing 72 hours. There were statistically signiifcant differences between the groups(104.91, P=0.00). RT-PCR results showed that with the increase of the concentration of 5-FU, theΔ133p53 mRNA, MDM2 mRNA and cyclin G1 mRNA expression gradually declined in gastric cancer cell line MKN45 cells, and there were statistically signiifcant differences between the groups(F=738.532, 1 396.607, 2 785.56,P=0.00). Cor-relation analysis showed that the expressions ofΔ133p53 mRNA and MDM2 mRNA in gastric cancer were positively correlated (r=0.871, P=0.01), while the expression of cyclin G1 mRNA and p53 mRNA had no obvious relevance (P=0.13). Conclusion:In the 5-FU-MKN45 gastric cancer cell line model, anti-tumor pathway ofΔ133p53 isomers is related with MDM2 but was not related with cyclin G1.

Background and purpose: Little about the function of p53 isoforms in gastric cancer was reported. This study was designed to explore the role ofΔ133p53 in the effect of recombinant mutant human tumor necrosis factor (rmhTNF) on gastric cancer cells, and provide a new basis for the diagnostics and therapeutics of gastric carcinoma. Methods: MKN45 (withΔ133p53 expression) or SGC7901 (withoutΔ133p53 expression) cells were treated with rmhTNF of different concentrations only or combined with 5-FU (a traditional gastric cancer cellular killer), and the growth inhibition rate and apoptosis was detected by CCK-8 and lfow cytometry. mRNA expressions ofΔ133p53, Gadd45αand CyclinB1 were measured by nested reverse transcription-polymerase chain reaction (nRT-PCR) or real-time polymerase chain reaction(RT-PCR). Results:On MKN-45 cells with positiveΔ133p53 expression, the inhibitory effect of rmhTNF was signiifcant, the inhibition rates of 50 and 500 IU/mL rmhTNF were 24.82%, 72.33%after culturing for 24 h (t=-9.558, P<0.01);also, the inhibitory effect of 5-FU was improved by rmhTNF remarkably in time-and dose-dependence, the inhibition rates of 5-FU (25 μg/mL), rmhTNF (50 IU/mL) combined with 5-FU (25 μg/mL), rmhTNF (500 IU/mL) combined with 5-FU (25 μg/mL) were 18.20%, 48.66%, 59.83%, separately, after culturing for 24 h (F=82.742, P<0.01);the inhibition rates of rmhTNF (50 IU/mL) combined with 5-FU (25 μg/mL) were 48.66%, 68.20%, 85.23%, separately, after culturing for 24 h, 48 h and 72 h (F=128.583, P<0.01). However, on SGC-7901 cells with negativeΔ133p53 expression, no growth inhibition was showed by rmhTNF only, the inhibition rates of 50 and 500 IU/mL were 2.74%, 3.25%after culturing for 24 h (t=-0.121, P>0.05). In apoptosis test, the apopto-sis-enhancing effect of rmhTNF was signiifcant on MKN45 cells, and the apoptosis-enhancing effect of 5-FU was fur-ther promoted signiifcantly by rmhTNF, the apoptosis of rmhTNF (50 IU/mL), rmhTNF (50 IU/mL) combined with 5-FU (25 μg/mL), rmhTNF (500 IU/mL) combined with 5-FU (25 μg/mL) were 18.20%, 48.66%, 59.83%, separately, after culturing for 24 h (F=123.931, P<0.05). In mRNA measurement, down-regulation ofΔ133p53 and CyclinB1, up-regula-tion of Gadd45αwere signiifcant in MKN45 cells treated by rmhTNF alone or combined with 5-FU. In nRT-PCR anal-ysis, the mRNA levels ofΔ133p53 were relatively 0.886, 0.499, 0.330, 0.161 (F=240.927, P<0.01);In real-time PCR analysis, the mRNA levels of Gadd45αwere 1.227, 1.694, 3.394, and the mRNA levels of CyclinB1 were 1.221, 0.722, 0.316, relatively. The expression ofΔ133p53 was positively related to CyclinB1 (r=0.977, P<0.01), but negatively re-lated to Gadd45α(r=-0.950, P<0.01). Conclusion:InΔ133p53 positively expressed MKN45 cells, rmhTNF showed as an effective tumor inhibitor and an enhancer of 5-FU as well, and this effect might be helped by two p53 down-stream molecules CyclinB1 and Gadd45α. The results suggest thatΔ133p53 might be a key target for the biological effect of rmhTNF against gastric cancer.

Background and purpose:To date, it mainly depended on imaging examination for detection of residual lesions, recurrence and distant metastasis, evaluation the sensitivity of radiotherapy and chemotherapy, and prognosis in nasopharyngeal carcinoma (NPC). Thus, searching for new tumor markers for NPC early diagnosis and individualized treatment is still merited. This study was aimed to investigate the expressions of serum macrophage inflammatory protein (MIP)-3α and cystatin A in patients with NPC before and after treatment, and to explore two markers’ value in NPC diagnosis, clinicopathological characteristics and clinical outcome assessment. Methods:The serum levels of MIP-3αand cystatin A in 140 primary NPC patients without distant metastasis before and after treatment were detected by enzyme-linked immunosorbent assay (ELISA) and compared with those in 100 healthy controls. Results:The sensitivity of MIP-3αand cystatin A were 92.1%and 42.1%, respectively;and the specificity of MIP-3αand cystatin A were 86.0%and 85.0%, respectively. All 140 NPC patients had complete remission (CR) or partial remission (PR). Serum levels of MIP-3αand cystatin A in pre-treatment patients with NPC were higher than those in post-treatment patients and controls. Serum MIP-3αand cystatin A levels were associated with overall stage of NPC, and MIP-3αwas also associated with T classification of NPC. The serum MIP-3αlevel in NPC with CR after treatment reduced to the level in control group, and that was still significantly higher in NPC with PR than in control group. No significant difference was found in the serum cystatin A level between NPC with CR or PR after treatment and control group. During 1-year follow-up, the post-treatment serum levels of MIP-3αand cystatin A were significantly higher in patients with distant metastasis than in patients without distant metastasis and controls. There was found statistically significant correlation between MIP-3α and cystatin A.Conclusion:MIP-3α may be a potential marker of NPC serological diagnosis. The detection of serum MIP-3αand cystatin A may contribute to the NPC staging and prediction of short-term clinical outcomes.

Objective To confirm the hepatotoxicity of valproic acid (VPA ) and its metabolites (2-propyl-4-pentenoic acid ,3-hydroxy valproic acid ,5-hydroxy valproic acid) on human liver cells .Methods Cells were divided into control group and VPA-treated group .The control group was conventionally cultured while the VPA-treated group was treated with valproic acid and its metabolites . The rate of cell proliferation was assayed by CCK 8 protocol . The mRNA levels of CYP1A1 , CYP1A2 ,PCNA ,Bax and Bcl-2 were measured by real time PCR .The correlated protein levels were measured by Western Blotting .The activity of LDH ,AST and ALT were also detected .Results Compared to the control group ,with the increases of concentrations and reaction time of VPA and its metabolites ,the proliferation rate of L02-cell was reduced ,the mRNA and protein levels of CYP1A1 ,CYP1A2 ,and Bax was increased ,the mRNA and protein level of PCNA and Bcl-2 was decreased , AST ,ALT ,and LDH were also elevated in the treated group .Conclusion Valproic acid and its metabolites were positively re-lated to hepatotoxicity .

Objective To analyze the advantages and surgical experience of laparoscopic anatomical surgery for adrenal tumors.Methods A total of 156 patients with benign or malignant adrenal tumors admitted to our hospital from April 2012 to September 2016 were enrolled and underwent laparoscopic anatomical surgery in the study.The curative effect and advantages of the operation were analyzed,and surgical experiences of laparoscopic anatomical surgery for adrenal tumors were summarized.Results Laparoscopic adrenalectomy was successfully completed in 155 of the 156(99.4％)patients and 1 patient with pheochromocytoma underwent open surgery due to pneumothorax caused by an accidental diaphragm injury.Postoperative pulmonary embolism occurred in 1 case and hematoma in right adrenal region in 6 cases,all of whom were completely relieved after intervention.Postoperative surgical specimen of adrenal glands in 54 patients were histopathologically examined,the results of which showed the negative boundaries in all patients.Patients were followed up for a mean of 11.6 months(range,4 to 26 months).All patients had a good recovery after surgery without recurrence of malignant tumor in local or trocar site.Conclusions As compared with open surgery,the transperitoneal laparoscopic adrenal surgery provides a larger operation area for good orientation and vision,and has such advantages as shorter hospital stay,fewer postoperative complications and better prognosis in patients with malignant adrenal tumors,which is worthy of clinical selection.

OBJECTIVE: To study the effects of Wuzhi capsule/schisantherin A (SchA) combined with cyclophosphamide on the pharmacokinetics of cyclophosphamide (CTX) in rats. METHODS: A total of 36 rats were randomly divided into CTX group (via tail vein, iv, CTX solution 300 mg/kg), CTX+WZC group (ig, Wuzhi capsule 300 mg/kg+via tail vein, iv, CTX solution 300 mg/kg), CTX + SchA low-dose, medium-dose, high-dose and excessive high-dose groups (ig, SchA 30, 300, 3 000, 30 000 μg/kg+via tail vein, iv, CTX solution 300 mg/kg) with 6 rats in each group. Blood samples were collected from orbital venous plexus of rats before medication and 0.083, 0.25, 0.5, 1, 2, 3, 4, 6, 8, 12, 24, 36, 48 h after medication.UPLC-MS/MS method was applied for concentration determination of CTX and its metabolites [de-chloroethyl CTX (DC-CTX), 4-ketone CTX (4-keto CTX), carboxyl phosphamide (CPM)] in plasma of rats. The plasma concentration-time curve was obtained. The pharmacokinetic parameters were fitted by using DAS 2. 0 software. RESULTS: The maximum plasma concentration (cmax) of DC-CTX in CTX group, CTX+WZC group, CTX+SchA low-dose, medium-dose, high-dose and excessive high-dose groups were (22 167. 85 ±2 844. 93), (10 920. 53 ± 1 490. 89), (18 951. 29 ± 1 558. 81), (18 622. 08 ± 791. 19), (18 515. 20 ± 2 560. 61), (15 133. 21 ± 1 305. 07) μg/mL, respectively; the area under the curves (AUCo-48 h) were (173 864. 01 ± 65 342. 21), (100 996. 98 ± 33 530. 02), (137 028. 16 ± 45 975. 19), (131 650. 18 ± 53 196. 41), (113 699. 40 ± 34 131. 36), (110 773. 27 ± 30 307. 15) μg·mL/h, respectively. Compared with CTX group, cmax of DC-CTX in CTX group, CTX+SchA low-dose, medium-dose, high-dose and excessive high-dose groups were decreased by 50. 74％, 14. 51％, 16. 10％, 16. 48％, 31. 73％, respectively. AUC0-48 h were decreased by about 42. 23％, 21. 45％, 24. 63％, 33. 37％, 36. 55％, respectively; with statistical significance (P<0. 05). The pharmacokinetic indexes as t1/2, tmax had no significant change. CONCLUSIONS: To some degree, both WZC and SchA can reduce the generation of DC-CTX, which indicates both of them can inhibit CTX toxicity metabolism pathway so as to reduce the generation of toxic metabolite chloroacetaldehyde. The inhibitory effect of SchA on toxicity metabolism pathway is weaker than that of WZC, and does not have a dose-dependent inhibitory effect.

In recent years , many studies have found that vitamin D insufficiency is associated with osteomalacia , hyperten-sion, diabetes, metabolic syndrome and other diseases .Vitamin D must be hydroxylated in the liver by a 25-hydroxylase for the first time, and then in the kidney by a 1α-hydroxylase for the second time to form the active metabolite 1,25-dihydroxy vitamin D ,which binds to the intracellular vitamin D receptor and exerts its effects .This paper reviewed the relationship between vitamin D and disease , present research situation of gene polymorphism of vitamin D receptor , and the advantages and limitations of several methods of vitamin D detection, and proposed the best method for detecting vitamin D receptor gene polymorphism and the importance of the detection state to guide clinicians to use drug rationally .