OBJECTIVE:To prepare nifedipine (NF) hollow controlled-release microspheres and evaluate the quality. METH-ODS:Solvent diffusion volatilization method was used to prepare microspheres,using comprehensive scores of cumulative release in 2,12,24 h(Q2 h,Q12 h,Q24 h)as indexes,orthogonal test was designed to screen the carrier material ethyl cellulose(EC),poly-vinyl pyrrolidone(PVP)and main drug NF amounts;appearance,particle size distribution,drug loading,floating and cumulative release of the microspheres prepared by optimal formulation were evaluated and compared of in vitro release behavior with imported preparation of Nifedipine controlled-release tablets (Adalat?). RESULTS:The optimal formulation was as follow as NF 3.00 g, PVP 1.60 g,EC 15.65 g. Prepared NF hollow controlled-release microspheres were spherical in shape with particle size distribution of 24-40 mesh and drug loading of 8.66%;24 h floating rate in release medium was 97.93%,Q2 h,Q12 h,Q24 h were 20.49%, 52.90%,91.00%(RSD<10%,n=3). Compared with the imported preparation,similarity factor f2 values of cumulative release were higher than 50,showing in vitro drug-release was consistent with the zero-order kinetic equation (r=0.9993);n of Rit-ger-Peppas equation (r=0.9807) was 0.478. CONCLUSIONS:Prepare NF hollow controlled-release microspheres show similar drug-release behavior with the imported preparation,the drug is released by the combination of diffusion and erosion.

According to the amino acids sequence of OC-IdeltaD86 gene and Escherichia coli codon usage, we synthesized this gene by overlap extension PCR method with 7 oligonucleotides DNA fragments. The PCR fragment was inserted into pGEM-T-easy vector and the recombined plasmid was named pGEM-T-OC-IdeltaD86. Two oligonucleotides into which the BamH I and Xho I sites were introduced were designed and synthesized based on pGEM-T-OC-IdeltaD86 and pet21b, and the PCR fragment into which the BamH I and Xho I sites were introduced was obtained. After digesting it with BamH I and Xho I, OC-IdeltaD86 gene was cloned into the corresponding sites of pet21b and obtained prokaryotic expression vector pet21b-OC-IdeltaD86. OC-IdeltaD86 gene was expressed in E. coli (BL21(DE3)plysS) after IPTG(Isopropyl beta-D-1-thiogalactopyranoside) inducement for 5 hours. The fusion protein of OC-IdeltaD86:6His gene accounted for 11.4% of total protein and 16.4% of soluble protein, which had been successfully purified by Ni-NTA and concentrated by PEG20000. This protein can effectively inhibit papain activity in vitro and may be used in anti-nematode research in vivo.
Cloning, Molecular ; Cystatins ; biosynthesis ; genetics ; Cysteine Endopeptidases ; metabolism ; Cysteine Proteinase Inhibitors ; genetics ; Escherichia coli ; genetics ; metabolism ; Genes, Plant ; genetics ; Mutation ; Oligonucleotides ; chemical synthesis ; genetics ; Oryza ; genetics ; Papain ; antagonists & inhibitors ; Prokaryotic Cells ; metabolism ; Recombinant Fusion Proteins ; biosynthesis ; genetics ; metabolism

Objective To study the effect of mental intervention on patients with postsurgical gastroparesis syndrome (PGS). Methods 70 patients with gastroptroparesis syndrome after abdominal operation were divided into the control group and the experimental group with 35 patients in each group according to chronological order.The patients in the control group accepted routine care,and the experimental group was given mental intervention based on routine care.Center for Epidemiological Survey Depression Scale (CESD) was used to evaluated the emotional state before and after intervention.Besides,the recovery of gastrointestinal function was appraised. Results The improvement level of anxiety in the experimental group was higher than the control group.The extinction time of symptoms,indwelling time of gastric tube,recovery time of food intake were shorter than the control group.Drainage time when gastric juice ＞ 800ml/d was shorter,recovery time of PGS and hospitalization time were shorter,the treatment cost was reduced,compared with the control group. Conclusions Mental intervention can alleviate the negative emotion and shorten the recovery time of patients with postsurgical gastroparesis syndrome.

Background: Bovine papilloma is a neoplastic disease caused by bovine papillomaviruses (BPVs), which were recently divided into 5 genera and at least 24 genotypes. Objectives: The complete genome sequence of BPV type 15 (BPV Aks-02), a novel putative BPV type from skin samples from infected cows in Southern Xinjiang China, was determined by collecting warty lesions, followed by DNA extraction and amplicon sequencing. Methods: DNA was analyzed initially by polymerase chain reaction (PCR) using the degenerate primers FAP59 and FAP64. The complete genome sequences of the BPV Aks-02 were amplified by PCR using the amplification primers and sequencing primers. Sequence analysis and phylogenetic analysis were performed using bio-informatic software. Results: The nucleotide sequence of the L1 open reading frame (ORF) of BPV Aks-02 was 75% identity to the L1 ORF of BPV-9 reference strain from GenBank. The complete genome consisted of 7,189 base pairs (G + C content of 42.50%) that encoded 5 early (E8, E7, E1, E2, and E4) and 2 late (L1 and L2) genes. The E7 protein contained a consensus CX2CX29CX 2 C zinc-binding domain and a LxCxE motif. Among the different members of this group, the percentages of the complete genome and ORFs (including 5 early and 2 late ORFs) sequence identity of BPV Aks-02 were closer to the genus Xipapillomavirus 1 of the Xipapillomavirus genus.Phylogenetic analysis and sequence similarities based on the L1 ORF of BPV Aks-02 revealed the same cluster. Conclusions The results suggest that BPV type (BPV Aks-02) clustered with members of the Xipapillomavirus genus as BPV 15 and were closely related to Xipapillomavirus 1.

The spectrum of biopsy-confirmed kidney disease varies with regions and periods. We describe the distribution of pathological types and epidemiological characteristics of kidney diseases in Northwest China due to regional differences in geographical environment, social economy, and dietary habits. Methods: Kidney biopsy cases from 2005 to 2020 in Xijing Hospital were retrospectively analyzed. Pathological characteristics of patients in different periods were analyzed using the t test or chi-square test. Joinpoint regression was used to analyze trends in pathological types and disease spectrum. Results: A total of 10,528 eligible patients were included. Primary glomerular disease (PGD) accounted for the majority of the cases and exhibited an obvious downward trend, whereas secondary glomerular disease (SGD) showed an obvious upward trend. Among PGD, immunoglobulin A nephropathy (IgAN) remained the most common pathological type, and the detection rate of membranous nephropathy (MN) was significantly increased. Among SGD, Henoch-Schönlein purpura nephritis (HSPN) was the most common pathological type and may present a significant characteristic of Northwest China. Diabetic nephropathy (DN) exhibited the most obvious upward trend in the whole process, whereas the fastest growth since 2012 was in hypertensive nephropathy. Conclusion: The proportion of SGD increased whereas PGD declined. IgAN remained the most common PGD, and HSPN was the most common SGD. MN and DN showed the most obvious upward trend among PGD and SGD, respectively. Changes in the spectrum of kidney disease, especially the constituent ratio of SGD, pose a great challenge to public health.

Objective:To investigate the influencing factors of recurrence after radical resection of middle and low rectal cancer, and to establish a prediction model based on magnetic resonance imaging (MRI) measurement of perirectal fat content and investigate its application value.Methods:The retrospective cohort study was constructed. The clinicopathological data of 254 patients with middle and low rectal cancer who were admitted to Tianjin Union Medical Center from December 2016 to December 2021 were collected. There were 188 males and 66 females, aged (61±9)years. All patients underwent radical resection of rectal cancer and routine pelvic MRI examina-tion. Observation indicators: (1) follow-up and quantitative measurement of perirectal fat content; (2) factors influencing tumor recurrence after radical resection of middle and low rectal cancer; (3) construction and evaluation of the nomogram prediction model of tumor recurrence after radical resection of middle and low rectal cancer. Measurement data with normal distribution were represented as Mean± SD, and measurement data with skewed distribution were represented as M(rang) and M( Q1, Q2). Count data were described as absolute numbers. Univariate and multivariate analyses were conducted using the COX regression model. The rms software package (4.1.3 version) was used to construct the nomogram and calibration curve. The survival software package (4.1.3 version) was used to calculate the C-index. The ggDCA software package (4.1.3 version) was used for decision curve analysis. Results:(1) Follow-up and quantitative measurement of perirectal fat content. All 254 patients were followed up for 41.0(range, 1.0?59.0)months after surgery. During the follow-up period, there were 81 patients undergoing tumor recurrence with the time to tumor recurrence as 15.0(range, 1.0?43.0)months, and there were 173 patients without tumor recurrence. The preoperative rectal mesangial fascia envelope volume, preoperative rectal mesangial fat area, preoperative rectal posterior mesangial thickness were 159.1(68.6,266.5)cm3, 17.0(5.1,34.4)cm2, 1.2(0.4,3.2)cm in the 81 patients with tumor recurrence, and 178.5(100.1,310.1)cm3, 19.8(5.3,40.2)cm2 and 1.6(0.3,3.7)cm in the 173 patients without tumor recurrence. (2) Factors influencing tumor recurrence after radical resection of middle and low rectal cancer. Results of multivariate analysis showed that poorly differentiated tumor, tumor pathological N staging as N1?N2 stage, rectal posterior mesangial thickness ≤1.43 cm, magnetic resonance extra mural vascular invasion, tumor invasion surrounding structures were independent risk factors of tumor recurrence after radical resection of middle and low rectal cancer ( hazard ratio=1.64, 2.20, 3.19, 1.69, 4.20, 95% confidence interval as 1.03?2.61, 1.29?3.74, 1.78?5.71, 1.02?2.81, 2.05?8.63, P<0.05). (3) Construction and evaluation of the nomogram prediction model of tumor recurrence after radical resection of middle and low rectal cancer. Based on the results of multivariate analysis, the tumor differentiation, tumor pathological N staging, rectal posterior mesangial thickness, magnetic resonance extra mural vascular invasion, tumor invasion surrounding structures were included to construct the nomogram predic-tion model of tumor recurrence after radical resection of middle and low rectal cancer. The total score of these index in the nomogram prediction model corresponded to the probability of post-operative tumor recurrence. The C-index of the nomogram was 0.80, indicating that the prediction model with good prediction accuracy. Results of calibration curve showed that the nomogram prediction model with good prediction ability. Results of decision curve showed that the prediction probability threshold range was wide when the nomogram prediction model had obvious net benefit rate, and the model had good clinical practicability. Conclusions:Poorly differentiated tumor, tumor pathological N staging as N1?N2 stage, rectal posterior mesangial thickness ≤1.43 cm, magnetic resonance extra mural vascular invasion, tumor invasion surrounding structures are independent risk factors of tumor recurrence after radical resection of middle and low rectal cancer. Nomogram prediction model based on MRI measurement of perirectal fat content can effectively predict the probability of postoperative tumor recurrence.

Objective:To explore whether endoscopic surgery can achieve the same effect on thyroid cancer as open surgery.Method:44 cases were selected to endoscopic thyroidectomy with breast areola approach, who were prepared to accept the surgical treatment of thyroid cancer with cT1N0 stages, female, ≤60 years old, no history of neck surgery and beauty desire. Fifty patients with the same conditions undergoing thyroid open surgery were as the control group.Results:Operative time of endoscopic group[ (170.0±28.0) min] was longer than that of the open operation group[ (90.0±21.0) min ( t=15.610, P=0.000) ]. There was no significant difference between the two groups in surgical complications ( P>0.05) . There was no statistical difference between the number of paratracheal and anterior tracheal lymph nodes of endoscopic group (6.0 ±4.2) and open surgery group (5.5 ±3.7) ( t=0.692, P>0.05) . There was no statistical difference between the number of anterior laryngeal lymph nodes of endoscopic group (0.7 ±1.1) and open surgery group (0.5 ±0.9) ( t=1.186, P>0.05) . Conclusion:The breast areola approach endoscopic thyroidectomy is safe and reliable in treatment of thyroid cancer, and the central cervical lymph node dissection is sufficient, which can be used as the choice of operation mode for thyroid cancer patients in cT1N0 stage.

Objective:To explore the application of pyrophosphorolysis-activated polymerization(PAP)to monitor plasma cfDNA in ad-vanced non-small cell lung cancer(NSCLC).Methods:A total of 85 patients diagnosed with advanced NSCLC between March 2016 and June 2017 were enrolled in the present study. EGFR mutations in cfDNA extracted from the plasma were detected using PAP and ARMS-PCR technology.The concordance analysis of EGFR mutations involved plasma vs.tumor tissue and PAP vs.ARMS-PCR.Further-more,38 EGFR-positive patients were selected to monitor EGFR mutations with PAP.Results:No statistical differences in EGFR muta-tions were observed between plasma and tumor tissue(P=0.092),as well as PAP and ARMS-PCR(P=0.210).The detection rate of EGFR mutations in cfDNA was higher in the progressor than in the non-progressor(62.5% vs.21.3%,P<0.001).Conclusions:PAP can be used for detecting and monitoring EGFR mutations in cfDNA to predict disease progression.

Objective:To explore the effects of radiation on miRNA-21 expression and the biology of human salivary gland adenoid cystic carcinoma cell lines(SACC-83,SACC-LM).Methods:In vitro cultured SACC-83 and SACC-LM cells were radiated by Varian 23 EX with the dose of 0(control),2,6 and 10 Gy respectively.Cell proliferation,apoptosis and miRNA-21 expression were observed by CCK-8 assay,flow cytometry and qRT-PCR methods respectively.Results:48 h after radiation the proliferation ability of SACC-LM cells decreased with the increasing dose of radiation(F =1 321.646,P =0.000),so did the miRNA-21 expression (F =177.964,P =0.011).However,the early apoptosis rate (F =354.484,P =0.039),the later apoptosis rate (F =254.278,P =0.042) and heteroploid ratio(F =1 562.991,P =0.001) increased with the increasing dose of radiation.In SACC-83 cell line 48 h after 6 Gy radiation the cell proliferation(F =1 537.214,P =0.013) and the miRNA-21 expression(F =134.868,P =0.017) were lower than that of other radiation doses.Moreover,the early apoptosis rate (F =88.579,P =0.006),the late apoptosis rate (F =1 391.345,P =0.033),heteroploid ratio(F =250.461,P =0.004) were higher than that of other radiation doses.Conclusion:The miRNA-21 expression in SACC-LM and SACC-83 cell lines is conversely associated with the radiation sensitivity.

Objective To study the effect of DNA-PKcs blocking on the expression of autophagic proteins,and proliferation of esophageal squamous cell carcinoma cells(EC109)after irradiation(X-Ray).Methods NU7441 was used to inhibit DNA-PKcs and X-Ray radiation treatment were used to treat EC109 cells.Th experiment was divided into 4 groups,including control group, NU7441 group,X-Ray group,X-Ray+ NU7441 group.The expressions of autophagy protein Beclin-1 and LC3B were detected by Western blot.Apoptosis was detected by flow cytometry.MTT assay was used to detect cell proliferation.Results The expression of p-DNA-PKcs in EC109 cells was decreased after NU7441 treatment and increased after X-ray irradiation.Compared with untreat-ed cells(control group),the expressions of both Beclin-1 and LC3B in X-Ray+NU7441 group were increased.Compared with the X-Ray group,the expression of Beclin-1 in the X-Ray+NU7441 group was increased.Compared with the control group,the apoptotic rate of EC109 cells in the X-Ray group and X-Ray+NU7441 group was significantly increased,the difference was statistically significant(P<0.05).Compared with the X-Ray group the apoptosis rate in the X-Ray-NU7441 group was significantly increased.The MTT results showed that compared with the control group,the proliferation in the X-Ray group and X-Ray+NU7441 group was significantly inhibited, the difference was statistically significant(P<0.05).Conclusion NU7441 inhibits the expression of DNA-PKcs protein in EC109 cells, which could promote the expressions of autophagy protein Beclin-1 and LC3B,promotes apoptosis and inhibits cell proliferation.