Objective:To analyze the genetic mutation characteristics of glucose-6-phosphate dehydrogenase (G6PD) deficiency among infants in Kunming.Methods:A total of 15 533 infants (7 994 males and 7 539 females) born in Kunming from January 1, 2018, to December 31, 2020, with an age range of 2 to 44 days, were selected. G6PD enzyme activity and gene mutation types were detected using fluorescence quantitative analysis, multicolor melting curve analysis (MMCA), and Sanger sequencing. Droplet digital PCR (ddPCR) was used for quantitative analysis of a newly identified variant family to determine the mutant allele proportion in family members. Meanwhile,the protein structure model and pathogenicity prediction of the novel variant were analyzed.Data analysis was conducted using SPSS 26.0. Specifically, chi-square tests were used for the detection rates of G6PD enzyme activity and gene mutations between different genders. One-way analysis of variance (ANOVA) was used for the comparison of enzyme activity among different mutation types.Results:Among 15 533 infants, 143 cases (129 males and 14 females) were tested positive for G6PD activity, with a detection rate of 0.92% (143/15 533). The difference in detection rates between males and females was statistically significant (χ 2=96.76, P<0.001). Out of 89 enzyme activity-positive cases (83 males and 6 females) underwent genetic testing, 77 (72 males and 5 females) were detected by MMCAand other 12 negative samples were underwent further Sanger sequencing, revealing mutations in 6 samples, all of which were males. Among the 83 individuals with gene mutations, 78 had heterozygous mutations, 1 had a homozygous mutation, and 4 had compound heterozygous mutations. A total of 12 mutation types were detected, with G6PD c.487G>A, c.1024C>T, c.1388G>A, and c.1376G>T being the most common, accounting for 74.70% (62/83) of all mutation types. The average G6PD enzyme activity of c.1376G>T was the lowest, and the differences were statistically significant compared to the average enzyme activity of the other three mutations ( P<0.05). One male infant with a newly identified G6PD c.242G>C mutation was detected, predicted to be pathogenic. ddPCR confirmed that the mother of the affected child was a c.242G>C mutant chimera, with a chimera proportion of 6.66%. Conclusions:In the Kunming region, the predominant G6PD deficiency gene mutation is c.487G>A, with the detection of a novel G6PD c.242G>C mutation. The application of ddPCR technology can assist in detecting the proportion of mutation chimeras.

Hepatectomy and liver transplantation are the most effective radical treatment for patients with hepatocellular carcinoma, but the high recurrence rate after surgery which seriously affects the prognosis of patients cannot be ignored. Microvascular invasion (MVI) is a risk factor for postoperative recurrence and metastasis in patients with hepatocellular carcinoma. There is no consensus or guideline recommendation locally or intermutually on postoperative adjuvant therapy of patients with hepatocellular carcinoma with MVI. Appropriate selection of postoperative adjuvant therapy is worth more in-depth discussion. This article reviews recent and relevant studies on postoperative adjuvant therapy for patients with hepatocellular carcinoma and MVI, including local anti-tumor therapy, systemic chemotherapy, immunotherapy, targeted therapy and combination therapy, with the aim to provide better reference to clinicians in managing these patients with postoperative adjuvant therapy.

Objective: To explore the potential genetic target of Stephania terandra in the therapy for hypertension based on inte-grated pharmacology platform to study the molecular mechanism of Chinese medicines in treating hypertension. Methods: Using the functional prediction modules of integrated pharmacology platform and the functional modules of network construction, the component-target network of Stephania terandra was calculated, and the target network of anti-hypertension was further calculated, and the target molecules related to hypertension were explored. Results: A total of 198 targets related to hypertension were obtained from the hyper-tension prevention. Among them, there were 59 potential action nodes and 20 known action nodes. The nodes with direct action were type-2 angiotensin Ⅱ receptor ( AGTR2), type-1 angiotensin Ⅱ receptor ( AGTR1), prostacyclin receptor ( PTGIR), solute carrier family 12 member 3 (SLC12A3) and endothelin-1 (EDN1). The GO enrichment and KEGG pathway enrichment of Stephania terandra chemical components showed a close correlation with hypertension, and the molecular mechanism of the target was clear. Conclusion:Through the integration of pharmacology platform, the targets of Stephania terandra-disease are deeply explored, and its potential mo-lecular mechanism as well as potential factors is studied.

'Three-acupoint and Five-needle Method' is a summary of Prof. Shao Jing-ming's clinical experience in the treatment of asthma for years. It is used to alleviate asthma during seizure and to improve the pulmonary functions, strengthen the body constitution and prevent recurrence in the remission stage. This article introduces the theoretic source and basic principle, and concrete operating method, i.e. puncturing bilateral Feishu (BL 13), Dazhui (GV 14) and bilateral Fengmen (BL 12), and adding acupoints based upon pattern identification, in combination of moxibustion and cupping therapy for promoting the clinical application of this method.

Objective To construct rCB1 gene eukaryotic expression vector, detect its expression in the cell, and explore its influence on apoptosis in human cervical cancer CaSki cells.Methods The total RNA was extracted from rat brains.The rCB1gene was amplified by RT-PCR.The pcDNA3.1(+)-rCB1 was constructed by enzyme digestion, purifi-cation, bind the PCR purification products and pcDNA3.1 (+) DNA.The pcDNA3.1 (+)-rCB1 plasmid was transfect-ed into HEK293 and CaSki cells by liposomes.The expression and localization of rCB1 were detected by Western blot and immunofluorescence combined with confocal laser scanning microscopy.The apoptosis rate of CaSki cells was detected by flow cytometry.The expression of rCB1, Bcl-2, Bax and Bad was detected by Western blot and real-time fluorescence quantitative RT-PCR (qRT-PCR).Results The 5300 bp pcDNA3.1(+) and 1500 bp rCB1 were obtained after diges-ting the pcDNA3.1 ( +)-rCB1.The result of sequencing was in agreement with the sequence of rCB1 gene ( NM_012784.4 ) .The rCB1 expressed in the membrane and cytoplasm when pcDNA3.1 (+)-rCB1 plasmid was transfected into HEK293 cells.The apoptosis rate of rCB1 group was increased compared with the blank group when pcDNA3.1 (+)-rCB1 plasmid was transfected into CaSki cells (P<0.05).Compared with the blank group, rCB1 gene upregulated the expres-sion of Bax and Bad, and suppressed the expression of Bcl-2.The statistical difference was significant ( P <0.05). Conclusions The pCDNA3.1(+)-rCB1 eukaryotic expression vector is constructed successfully.It is found that rCB1 is expressed in membrane and cytoplasm of HEK293 cells.rCB1 can significantly promote the apoptosis in cervical cancer CaSki cells by up-regulating the expression of Bax and Bad, and down-regulating the expression of Bcl-2 as well.

OBJECTIVETo compare the application valuable of flexible spatial scan statistics and kulldorff scanning window in the cluster detection and early warning of hepatitis A.

METHODSThe case numbers and incidence data of hepatitis A in 2012 for all the counties (cities, districts)in Yunnan province were collected from China Information System for Disease Control and Prevention and the total number was 1 335. By extracting the time length by month, the flexible spatial scan statistics was tested by retrospective analyses of hepatitis A data in Yunnan in 2012 and compared the results with the Kulldorff circular scan statistic analyses.

RESULTSThe results of flexible scanning window showed that there were fifteen hepatitis A spatial clusters in Yunnan province in 2012 and in July, these areas including Gejiu county, Mengzi county and Wenshan county had the strongest clusters (the log likelihood ratio (LLR) = 52.66, P = 0.001). The results of Kulldorff scanning window showed that there were twenty hepatitis A spatial clusters and these areas including Gejiu county, Hekou county, Maguan county, Mengzi county, Pingbian county, Wenshan county had the strongest clusters (LLR = 47.82, P = 0.001). The results of the flexible scanning window were the same as the actual monitoring results. But the results of Kulldorff scanning window showed that in May and June some areas without incidence had the clusters.

CONCLUSIONFlexible scanning window can detect the monthly clusters of the Hepatitis A. Flexible scanning window had a higher accuracy than Kulldorff irregular circular scanning window. Flexible spatial scan statistics had the value in the use of spatial aggregation detecting on hepatitis A.

China ; Cluster Analysis ; Data Interpretation, Statistical ; Hepatitis A ; Humans ; Incidence ; Retrospective Studies

Objective To establish a simple, stable, specific and sensitive method for detection of Tyzzer ’ s or-ganism by reverse dot blotting ( RDB) .Methods Primers and specific probes were designed according to the conservative sequence of Tyzzer 16S rDNA.The forward primer was labeled with biotin .The reverse dot blotting method was established followed by PCR amplification .The specificity and sensitivity of this method were determined .Next, 41 mice and 38 rats were examined by RDB , ELISA and IFA .Results The RDB method showed a high specificity , and in the testing of the 79 laboratory animals , its limit of detection was 4.5 ng/μL.Compared the results of ELISA and IFA , its consistence with ELISA was 100%and the positive rate was 7.59%(6/79), the consistence with IFA was 92.4%(73/79), and the posi-tive rate was 0%.Conclusions An accurate, sensitive and specific method in combination with PCR and RDB in detection of Tyzzer’ s organism is established in this study .

Objective To explore the protective effect of Sijunzi decoction and Danggui Buxue decoction on the immunological function in tumor-bearing mice treated by cyclophosphamide.Methods Transplantable carcinomas animal model were constructed by inoculating tumor cell to mice for 24 hours,and randomly divided into five groups.Tumor-bearing control group were reveived normal saline 0.2 mL per mice one day by intraperitoneal injection,and normal saline 0.4 mL per mice each day by intragastric administration.Cyclophosphamide group were reveived CTX 20 mg per kg weight each day,and normal saline 0.4 mL per mice each day.Sijunzi decoction and Danggui Buxue Decoction low dose group were reveived CTX 20 mg per kg weight each day and traditional Chinese medicine 0.1 g per mice each day.Sijunzi decoction and Danggui Buxue Decoction mediate dose group were reveived CTX 20 mg per kg weight each day and traditional Chinese medicine 0.2 g per mice each day.Sijunzi decoction and Danggui Buxue Decoction high dose group were reveived CTX 20 mg per kg weight each day and traditional Chinese medicine 0.4 g per mice each day.The weight indices of thymus gland and spleen,NK-cell activity and the proliferation of T-lymphocyte in five groups were measured after ten 10 days treatment.Results The weight indices of thymus gland and spleen,NK-cell activity and the proliferation of T- lymphocyte were increased in all three Sijunzi decoction and Danggui Buxue decoction groups,especially in Sijunzi decoction and Danggui Buxue decoction mediate and high dose group,the differences between them and CTX group were statistically significan(P<0.05,P<0.01).Conclusion Sijunzi decoction and Danggui Buxue decoction can antagonize the decrease of immunological function in tumor-bearing mice treated by chemotherapy.

OBJECTIVETo generate a mouse model of chronic hepatitis B (CHB) infection by performing in vivo transduction of hepatitis B virus (HBV) covalently closed circular (ccc)DNA.

METHODSNude mice were injected with HBV cccDNA at doses of 1.5, 1.0 or 0.5 mug/ml. A control group was generated by giving equal injection volumes of physiological saline. The serum levels of hepatitis B surface antigen (HBsAg) and hepatitis B e antigen (HBeAg) on post-injection days 1 and 3, weeks 1-6, 8 and 10 were assayed by reflection immunoassay. At post-injection week 10, all animals were sacrificed and liver tissues were collected. Copies of HBV DNA in serum and liver tissue were detected by real-time PCR. HBV antigens in liver tissue were detected of by immunohistochemistry. Pathological analysis of liver tissue carried out with hematoxylin-eosin staining. Linear correlation of data was determined by statistical analysis.

RESULTSHBsAg and HBeAg were detected in sera from all three groups of cccDNA-injected mice staring at post-injection day 1 and lasting through week 10. The levels of HBsAg over the 10-week period showed two patterns of increase-decrease;the lowest level was detected at week 4 and the highest level was detected at week 8. In contrast, the levels of HBeAg over the 10-week period showed three patterns of increase-decrease; the lower levels were detected at weeks 2 and 4 and the higher levels at weeks 3 and 6. HBV DNA copies in liver tissues showed a cccDNA dose-dependent descending trend over the 10-week study period (1.5 mug/ml:1.14E+07 ± 6.51E+06 copies/g, 1.0 mug/ml:9.81E+06 ± 9.32E+06 copies/g, and 0.5 mug/ml:3.72E+06 ± 2.35E+06 copies/g; Pearson's r =0.979). HBV DNA copies in sera showed the pattern of 1.0 mug/ml cccDNA more than 1.5 mug/ml cccDNA more than 0.5 mug/ml cccDNA, and in general were higher than those detected in the liver tissues. Liver tissues from all cccDNA-injected mice showed positive immunohistochemistry staining for both HBsAg and HBeAg. HE staining showed that the liver tissues of all cccDNA-injected mice had severe fatty and vacuolar degeneration and less obvious structure of liver lobules (compared to the liver tissues from control mice).

CONCLUSIONThe CHB mouse model successfully established in this study by in vivo transduction of HBV cccDNA may represent a useful tool to study the pathogenic mechanisms and potential antiviral treatments of human CHB.

Animals ; DNA, Circular ; administration & dosage ; DNA, Viral ; administration & dosage ; Disease Models, Animal ; Hepatitis B Surface Antigens ; blood ; Hepatitis B e Antigens ; blood ; Hepatitis B virus ; genetics ; physiology ; Hepatitis B, Chronic ; virology ; Male ; Mice ; Mice, Nude ; Transduction, Genetic ; Virus Replication

Objective To compare the application valuable of flexible spatial scan statistics and kulldorff scanning window in the cluster detection and early warning of hepatitis A.Methods The case numbers and incidence data of hepatitis A in 2012 for all the counties( cities, districts) in Yunnan province were collected from China Information System for Disease Control and Prevention and the total number was 1 335.By extracting the time length by month, the flexible spatial scan statistics was tested by retrospective analyses of hepatitis A data in Yunnan in 2012 and compared the results with the Kulldorff circular scan statistic analyses.Results The results of flexible scanning window showed that there were fifteen hepatitis A spatial clusters in Yunnan province in 2012 and in July, these areas including Gejiu county, Mengzi county and Wenshan county had the strongest clusters(the log likelihood ratio(LLR) =52.66,P=0.001). The results of Kulldorff scanning window showed that there were twenty hepatitis A spatial clusters and these areas including Gejiu county, Hekou county, Maguan county, Mengzi county, Pingbian county, Wenshan county had the strongest clusters (LLR=47.82,P=0.001).The results of the flexible scanning window were the same as the actual monitoring results.But the results of Kulldorff scanning window showed that in May and June some areas without incidence had the clusters.Conclusion Flexible scanning window can detect the monthly clusters of the Hepatitis A.Flexible scanning window had a higher accuracy than Kulldorff irregular circular scanning window. Flexible spatial scan statistics had the value in the use of spatial aggregation detecting on hepatitis A.